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1.3: Micropipetting - Biology

1.3: Micropipetting - Biology


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Learning Objectives

Goals:

  • Use various instruments found in the biotechnology lab.
  • Measure volume with precision and accuracy.
  • Pipet with precision and accuracy.
  • Learn how to use a micropipette to measure very small volumes.

Student Learning Outcomes:

Upon completion of this lab, students will be able to:

  • Make accurate and precise measurements with micropipettes and serological pipettes.
  • Calculate percent error for a given measurement.
  • Read, set, and operate a micropipette.
  • Determine which pipette should be used to measure a specific volume.
  • Determine how accurately you can measure with each micropipette.

Introduction to Micropipetting:

The ability to measure very small amounts, microliters (µl), of liquid chemicals or reagents is a fundamental skill needed in the biotechnology or research lab. Scientists use a device called a micropipette to measure these very small volumes with accuracy. This activity introduces the technique of micropipetting. Remember, as with all fine motor skills, this new skill will require practice and determination. Be sure to operate the micropipette slowly and carefully.

Part I: Choosing and Setting the Micropipette

There are several sizes of micropipettes used in the biotechnology lab. Today, you will be using the P-1000, P-200, and P-20. The P-1000 measures volumes between 100-1000 µl, the P-200 measures volumes between 20-200 µl, and the P-20 measures volumes in the 2-20 µl range. It is important to always pick the correct micropipette for the volume to be measured.

Looking at Figure 3.1, you can see that each micropipette has a similar but different display window. For the P1000, the red number indicates the thousands place, followed by the hundreds, tens, and the ones displayed as small vertical lines. Each line represents 2 µl. The P-200 is reads differently. The display from the top down reads, hundreds, tens, ones, and the vertical lines are considered 0.2 µl. Finally, the P-20 can be read from the top down tens, ones, and the red tenths.

A. Choosing your Micropipette

For each amount listed below, indicate the correct micropipette needed to measure the volume accurately then set the pipette to the indicated amount and show your partner.

Table 1. Pipette Size Choice
AmountPipette NeededPartner Observation
  1. 567 µl:

2. 160 µl:

3. 700 µl:

4. 25 µl:

5. 15 µl:

B. Setting your Micropipette

Materials

  • P-20 micropipette
  • P-20 tips
  • Waste container
  • Tube of red dye in tube rack
  • Laminated sheet for pipetting

Procedure

  1. Each student will load 5, 10, 15, and 20 µl of red dye onto the laminated sheet.
  2. Locate the p-20 and set the dial to 5 µl.
  3. Hold the micropipette in your dominant hand, and gently but securely place the end of the micropipette into the proper size tip. Once the tip is on, be careful not to touch the tip on anything! If your tip touches the bench, lab coat etc. eject the tip into the waste container and get a new clean pipet tip.
  4. With your other hand, open the cap of the tube of red dye and bring the tube of red dye to eye level,
  5. Push the micropipette plunger down to the first stop and hold your thumb in this position.
  6. Place the pipet tip into the red dye solution.
  7. Gently release your thumb from the plunger to draw fluid into the tip.
  8. Confirm that the tip has liquid and that no bubbles are present within the tip.
  9. Close the tube of red dye and place back in tube rack.
  10. Gently touch the tip to the center of the circle labeled 5 µL and slowly push all the way down (to second stop) on the plunger to dispense the liquid.
  11. Repeat this process for the remaining volumes.
  12. Be sure to watch your groupmates to provide feedback and help with their technique.

Results

Take a picture or draw a picture of your spots and include this in your lab notebook as Figure 1. Make sure the figure has a title.

Conclusion

  1. Observe if your spots were similar in size to your groupmates.
  2. Which volume had the most variability?
  3. What could have contributed to your spot being too large or small?

Part II: Pipetting Practice

A. Microplate Art

Materials

  • p20 pipette (1)
  • p200 micropipette (1)
  • P-20/P-200 tips
  • Microplate art set (design cards, colored dyes, and 96-well microplate) (1)
  • Analytical or electronic balance

Procedure

  1. Obtain a 96-well microplate, a design card, and tubes of colored dyes.
  2. Write the Microplate Art Design number in your lab notebook.
  3. Using the gram balance, obtain the weight of your 96 well microplate and record in your notebook.
  4. Using the p200 micropipette with tip, dispense 50 µl of dye into the wells written on the design card.
  5. Once you have finished pipetting, weigh your completed microplate, and record in your lab notebook.

Results

  1. Be sure to record your weight in grams of your microplate pre/post pipetting in your lab notebook.
  2. Using these values, calculate your percent error of the microplate you just created. Include the calculation in your lab notebook.
  3. Take a picture of your microplate design and include this in your lab notebook.

CONCLUSION

  1. Was your percent error below +/- 5%? If your percent error was above this range, elaborate on the potential causes.
  2. Did your pattern look correct? How could you avoid errors in the future?

B. Micropipette Practice Matrix

Materials

  • p20 pipette (1)
  • p200 micropipette (1)
  • 1.5ml microfuge tubes (3)
  • Permanent marker
  • Analytical or electronic balance

Procedure

  1. Label three microfuge tubes: 1, 2, 3,
  2. Weigh each tube before placing any liquid inside.
  3. Draw table 2 in your lab notebook and use it to record your data.

Results

Table 2. Calculating Accuracy for Micropipetting

Tube #

Weight of tube (g)

Weight of tube + dye (g)

Theoretical weight of dye

Actual weight of dye

% Error

1

2

3

  1. Deliver the volumes indicated in Table 3 into each of the 3 labeled tubes.
Table 3. Volumes to be Pipetted into each Tube

Tube #

Micropipette

Red Dye (µl)

Blue Dye (µl)

Green Dye (µl)

1

P1000

210

435

332

2

P200

110

153

67

3

P20

15

17

10

  1. Weigh each tube after pipetting.
  2. Determine the theoretical weight of the dye using the information about the weight of a mL of the dye solution at room temperature provided by your instructor.
  3. Determine the % error for each tube.

Conclusion

Based on your data comment on the following in your lab notebook:

  1. Which micropipette gave the most precise measurement?
  2. Which micropipette gave the most accurate measurement?
  3. What may have contributed to higher percent errors?

Study Questions

  1. Convert the following:
    • 345 mL = __________________ µl
    • 0.54 mL = _________________ µl
    • 5.2 L = ________________ mL
  2. Which micropipette would you choose to measure 550µl? 17µl? 167µl?
  3. Make 3 suggestions that other biotechnologists can use to improve micropipetting accuracy.
  4. Assuming that the density of water is 1 gram per milliliter, how much should 550 µL of water weigh?
    • 17 µL of water?
    • 167 µL of water?
  5. What is the formula to calculate percent error?
  6. What is the maximum volume you can set for each micropipette (P-1000, P-200, P-20)?

RESNA Annual Conference - 2014

Approximately 20% of the working population in the United States has a disability (Dept. of Labor, 2013). Yet only 2.7% of the science, technology, engineering, and mathematics (STEM) workforce report having a disability (Miner, Nieman, Swanson, Woods, 2001). Additionally, less than 3% of all biological sciences doctorates are earned by persons with disabilities (PWDs) (Supalo, Mallouk, Amorosi, Lanouette, Wohlers & McEnnis, 2009 NSF, 2013). There are several factors in the educational experiences of PWDs which help explain their underrepresentation in STEM professions: lack of independent hands-on experiences, low expectations, lack of role models, and limited exposure to science in and out of the classroom (Dunn, Rabren, Taylor, & Dotson, 2012 Supalo, Mallouk, Amorosi, Lanouette, Wohlers & McEnnis, 2009). Active practical STEM learning experiences in the science classroom or research settings are crucial to the development and success of scientists (National Research Council, 1996, Stefanich, 2007). Indeed, when undergraduates with visual and mobility disabilities were interviewed, the deterrent to them pursuing a laboratory-based career was not a lack of interest but the lack and inaccessibility of hands-on learning experiences (Duerstock, 2006 Mansoor et al., 2010).

In order to provide practical research experiences for PWDs including persons with blind or low vision (BLV) disabilities, it is necessary that each of the procedural components of the research task be accessible. Due to the foreseeable needs of a BLV graduate student entering into a biology research field, we decided to look at the accessibility of two common molecular biology procedures: micropipetting and transferring cultured cells from a well plate to a microscope slide.

In a survey of Purdue University life science research faculty, 85% of respondents stated that micropipetting was performed daily in their labs. Micropipetting is a ubiquitous component of most molecular biology protocols, including cell culturing. Over half of survey respondents performed cell culturing at least 2 to 3 times a week. In this study, test experiments were conducted to demonstrate the proficiency of BLV researchers micropipetting and culturing cells on a microscope slide coverslip in a well plate and then transferring it to a slide. Micropipetting liquids into and out of the wells required positioning the micropipette tip in the approximate middle of the wells near the bottom without touching the coverslip. Though this procedure is commonly performed by researchers with normal vision and hand eye coordination, it is extremely difficult to do while blind. Additionally, transferring cultured cells from the microscope coverslip in the well plate to a slide is normally accomplished using forceps without damaging the cells. This requires fine dexterity and good eyesight. For our blind researchers, both micropipetting and removing the coverslip from the well plate with forceps was impossible, even disregarding aseptic techniques.

A micropipette guide placed on top of the six well plate was designed to direct the tip to the proper location and depth. In addition, the micropipette guide would protect against contamination of the wells containing cell cultures. In order to remove the coverslip from the bottom of the well, a holder was designed that fit in the bottom of the well plate with a protruding handle for easy removal and transfer. 3-D printing facilitated a rapid and inexpensive iterative design process of building, assessing, and redesigning prototype solutions.


Protocol

1. Prepare a Sterile Workspace

Before starting any sterilization procedures in your work area, wash your hands thoroughly with antiseptic soap and warm water.

Be sure to re-wash your hands any time you suspect you have contamination from your experimental manipulations.

Clear away all materials cluttering your work area on the laboratory bench. Remove a pre-moistened disinfectant wipe from the canister and wipe down the entire area. Allow the disinfectant to evaporate - do not wipe dry!

Use disinfectants such as alcohol (isopropanol or 70% ethanol) or phenolic compounds (o-phenylphenol) .

To prevent aerosolization, or the production of a fine mist containing bacterial cells, and spread of microbial contaminants, avoid dispensing disinfectant from a squeeze bottle.

Desiccation of microorganisms is one of the most effective ways to decontaminate surfaces.

Even if someone has recently used the laboratory bench and the bench top was wiped down with disinfectant, ALWAYS begin your laboratory time by wiping down the bench.

After the disinfectant has dried completely, use an igniter to light the Bunsen burner. Adjust the flame so that a blue cone can be seen in the middle of the flame. The flame is now producing an updraft, or air convection currents in which warm air rises up and away from the flame (Figure 1). As heat rises, microorganisms and dust particles are forced upward and away from the immediate work area. Work slowly, carefully, and deliberately at all times within this area created by the Bunsen burner, referred to as a sterile field. Keep the Bunsen burner on during the entire procedure.

The tip of the blue cone is the hottest part of the flame.

Be careful not to disturb the updraft by rapid movements that dramatically change the air currents around the laboratory bench. Creating an updraft with the Bunsen burner minimizes the possibility of microorganisms and dust falling onto the bench or into open bottles, tubes or flasks in the work area.

Arrange all the supplies needed for the procedure on the laboratory bench near the sterile field. Make sure all the materials are properly labeled.

Supplies may include serological pipettes and micropipettors, sterile culture tubes, sterile flasks, media bottles containing broth, sterile microcentrifuge tubes, micropipettor tips, racks for tubes, bacterial cell cultures, and phage stocks.

Liquid media should be sterilized in an autoclave at 121 ଌ for at least 15 minutes on the liquid setting. Larger volumes of media (> 1L) require longer autoclave times. Labware should be sterilized in an autoclave at 121 ଌ for at least 30 minutes on gravity (dry) setting.

In general, sterile solutions may be stored at 4 ଌ for up to 5 months. Note that storage time is significantly reduced for solutions containing unstable components such as antibiotics - always check the manufacturer's recommendations.

2. Transferring Liquids Using Serological Pipettes

Serological pipettes come in many sizes and options: plastic or glass, disposable or reusable, plugged or unplugged. These are calibrated to deliver volumes ranging from a 0.1 ml to 25 ml.

Common sizes for serological pipettes are 5 ml, 10 ml and 25 ml and should be used for aseptic liquid transfers of 0.1 ml or more (panel A of Figure 2). There also are larger serological pipettes that can deliver volumes up to 100 ml however, the focus of this protocol is on the more common, smaller sized pipettes.

Pre-sterilized pipettes with a cotton wool plug are needed for microbiology and tissue culture experiments. The plug should not be removed from the top of the pipette it is designed to function as a barrier to overfilling the pipette.

Different applications call for plastic versus glass serological pipettes. Glass is needed for organic solvents. Either can be used when performing BSL-1 experiments on the laboratory bench top. Only plastic may be used when working in a Biosafety cabinet with BSL-2 organisms where a Bunsen burner cannot be used. It also is recommended that plastic be used for applications involving transfer of molten agar.

Serological pipettes are of two types: TC ("to contain") or TD ("to deliver"). TC pipettes deliver all the volume, including the tip, and must be "blown out" or rinsed to get the specified volume. TD pipettes are calibrated to leave a tiny bit in the tip that should not be delivered. Be sure to check the label on the body of the pipette near the top to ascertain which type it is (Figure 3). The most commonly used are TD pipettes, which are marked with double rings at the top.

Take a sterile plastic serological pipette (also called a volumetric transfer pipette) and carefully remove the paper sleeve at the end with the cotton wool plug by peeling it away like the skin of a banana - do not remove the entire sleeve, protecting the tip of the pipette that will come in contact with the liquid to be transferred. Touch only the top of the pipette (above the graduation marks) with your hands.

Never go into a sterile solution with a used pipette, even if great care has been taken to keep it sterile.

Glass serological pipettes are typically stored in metal canisters (panel B of Figure 2). Loosen the top of the canister then carefully remove the cap, and flame the open ends of the cap and canister. Place the cap down, on its side, on the disinfected bench. Remove one pipette from the canister by holding it horizontally and gently shaking it so the tops of one or two pipettes stick out about an inch and can be easily grasped. Lay down the canister on its side and remove one pipette, but be cautious not to touch the other pipettes in the container. Do not touch the bottom tip of the pipette with your hands, and avoid contact of the tip with other non-sterile surfaces.

Affix a pipette aid such as a bulb, pump, or gun to the top end of the serological pipette. Remove the paper sleeve from the plastic pipette. Hold the pipette aid in your right hand.

If using a glass pipette, pass the bottom third of the pipette through the blue cone in the Bunsen burner flame for 1-3 seconds. Rotate the pipette 180° as it passes through the flame. Plastic pipettes and tubes cannot be flamed.

If left-handed, hold the pipette aid in your left hand and conduct subsequent manipulations of culture bottles and tubes with your right hand.

Contamination tends to occur with plastic pipettes when withdrawing the final inches of the pipette from the sleeve because the sterile tip comes into contact with the part of the sleeve touched by your hands.

Remove the cap of the bottle containing sterile media. Do not place the cap on the laboratory bench but hold it between your ring finger and palm of the right hand while manipulating the pipette aid with the thumb, index and middle fingers of the same hand (Figure 4). Holding the bottle at a 45° angle, pass the rim of the bottle through the flame of the Bunsen burner, creating a sterile field around the open bottle.

Although best avoided, if you must put the cap down, place it face down on a disinfected surface. With a cap that faces up, there is a greater chance of contamination from movements of objects or hands, creating air currents that cause microorganisms and dust particles to descend to the inside surface of the cap.

The purpose of flaming is not to sterilize but to warm the opening of the bottle and create air convection currents up and away from the opening (i.e., updraft). The warm, rising air helps prevent dust particles and other contaminants from entering the bottle.

Keep the sterile container open for as little time as possible. It is important to keeping the points of entry of airborne microorganisms to a minimum throughout the procedure.

Avoid coughing, sneezing, talking, and other inadvertent movement while sterile containers are open.

Never pass hands and fingers over the top of a sterile field (i.e., open bottles or flasks, the inside of tubes and bottle caps) once they have been passed through the Bunsen burner flame.

Always work with an open flame when opening sterile tubes or bottles. Never have more than one tube, bottle, or flask open on the bench at a time. Flaming should be done immediately upon opening and just before closing tubes, bottles and flasks.

Place the tip of the serological pipette into the bottle containing the sterile media then aspirate, or draw the sample aseptically, from the bottle. Use the pipette aid to control the flow of the sample into the pipette. Precisely read the volume drawn into the pipette by aligning the meniscus formed on top of the liquid column to the graduation marks on the calibrated pipette (Figure 5).

DO NOT MOUTH PIPET! Always use a pipet aid (pump, bulb, or gun).

Pay attention to the sequence of numbers when determining the volume aspirated. The numbers may be printed tip to top, or vice versa, or often times in both directions.

When reading the volume, always hold the pipette vertically, perpendicular to the ground, and view the liquid meniscus dead-on at eye level.

Serological pipettes are only as accurate as the smallest marked increment, which is typically 0.1 ml for 5 ml and 10 ml pipettes and 0.2 ml for 25 ml pipettes. If greater precision is needed, a serological pipette may be used in combination with a micropipettor.

Pass the rim of the bottle through the Bunsen burner flame once again, then place the cap back on the bottle. Set the media bottle aside.

Don't burn yourself with the Bunsen burner in a rush to close the bottle.

Hold a test tube or flask in your left hand. Remove and hold the cap as described in step #4 above. Create a sterile field by flaming the rim of the tube or flask in the Bunsen burner.

Dispense the media in the pipette into the tube or flask. Control the flow of the sample so it does not splash out of the tube or flask.

Volumes may be measured such that the entire volume is delivered and the pipette drains completely, or a specific volume is achieved by doing a point-to-point delivery (one volume marking to another).

Pass the rim of the tube or flask through the Bunsen burner flame once again, then replace the cap. Set the tube or flask aside. Remove the pipette aid, and discard the pipette into the proper waste receptacle.

Plastic serological pipettes are disposable, while glass serological pipettes can be sterilized and used again. Proper disposal requires plastic pipettes be placed in a designated sharps container (rigid box lined with plastic disposal bag) while glass pipettes initially should be immersed in a container with 10% bleach solution to disinfect the inside and outside surfaces. Then the glass pipettes should be thoroughly washed with laboratory detergent, rinsed with distilled water, and sterilized in an autoclave.

These same steps should be followed when inoculating media with a bacterial culture or phage stock or when performing serial dilutions.

3. Transferring Liquids Using Micropipettors

Precisely measuring and dispensing minute volumes can be accomplished using micropipettors (also referred to as Pipetman Panel A of Figure 6). These instruments come in different sizes each with a specific volume range: P2 for 0.2-2 μl , P10 for 1-10 μl, P20 for 2-20 μl, P200 for 20-200 μl, and P1000 for 200-1000 μl.

Treat micropipettors with care, as they are precision instruments. Do not leave them lying on the laboratory bench unattended where they can be knocked off and damaged. Do not allow pipettes to come into contact with corrosive chemicals.

For volumes greater than 1000 μl, use a serological pipette.

Although working within the sterile field created by the Bunsen burner, do not flame micropipettors, tubes and plastic tips. The tubes and tips should be pre-sterilized. The micropipettors may be wiped down with a pre-moistened disinfectant wipe prior to use.

A numeric volumeter showing the dispensed volume can be set by turning adjustment knob. Adjust the volume before proceeding to step #3.

NEVER TURN THE ADJUSTMENT KNOB ABOVE THE INTENDED RANGE!

To obtain maximum accuracy when decreasing the volume setting on the micropipettor, slowly dial down the thumb wheel, making sure not to overshoot the graduation mark.

To obtain maximum accuracy when increasing the volume setting on the micropipettor, dial the thumb wheel up, passing the desired graduation mark by 1/3 of a turn. Then slowly dial down the thumb wheel to reach the intended volume, making sure not to overshoot the graduation mark.

The volumeter shows three numbers. Depending on the micropipettor, the numbers are interpreted differently. Note that each micropipettor is only as accurate as the smallest graduation mark.P2: For volumes between 0.2-2.0 μl. The top number denotes volume in microliters. The second number indicates tenths of a microliter (0.1 μl), and the third number represents hundredths of a microliter (0.01 μl). Each graduation mark equals an increment of two one-thousandths (0.002 μl) of a microliter.P10: For volumes between 1.0-10.0 μl. The top number is for tens of microliters this usually is set at "0" and should only be set at "1" with the other two numbers set at "0" when dispensing 10.0 μl. The middle number denotes volume in microliters. The third number indicates tenths of a microliter (0.1 μl). Each graduation mark equals an increment of two one-hundredths (0.02 μl) of a microliter.P20: For volumes between 2.0-20.0 μl. The top number in black is for tens of microliters this should only be set at "2" with the other two numbers set at "0" when dispensing 20.0 μl. The second number in black denotes the volume in microliters. The third number in red indicates tenths of a microliter (0.1 μl). Each graduation mark equals an increment of two one-hundredths (0.02 μl) of a microliter.P200: For volumes between 20.0-200 μl. The top number is for hundreds of microliters this should only be set at "2" with the other two numbers set at "0" when dispensing 200 μl. The middle number indicates the dispensed volume in tens of microliters, and the third number denotes volume in microliters. Each graduation mark equals an increment of two one-tenths (0.2 μl) of a microliter.P1000: For volumes between 200-1000 μl. The top number is for thousands of microliters this usually is set at "0" and should only be set at "1" with the other two numbers set at "0" when dispensing 1000 μl. The middle number is for hundreds of microliters. The bottom number is for tens of microliters. Each graduation mark equals an increment of two (2 μl) microliters.

Performance check: These instruments should be calibrated annually, ensuring accuracy and precision are maintained to stay within ± 5% of specifications. Use an analytical scale to measure water, making sure the minimum and maximum settings correspond to the intended volume. For example, use a P1000 to transfers 200 μl of water to a weigh dish on the scale. Since water has a density of 1, then 1 ml of water is equivalent to 1 gram (g). Thus, 200 μl (0.2 ml) of water should be equal to 0.2 g. Also, make sure the tip does not leak and can maintain the desired volume until dispensed using plunger system.

Micropipettors must be used with plastic disposable tips at all times. Fit a tip tightly onto the end of the barrel of the micropipettor. Press down and twist slightly to ensure an airtight seal.

Tips are usually packed into plastic boxes that can be sterilized by autoclaving. Open the tip box to retrieve a tip, then close the tip box to minimize contact with contaminants in the air.

Some tips have filters similar to the cotton wool plugs on serological pipettes. These tips are often more expensive than regular tips and thus are used for specialized applications. For instance, when measuring volatile chemicals such as chloroform or radioactive liquids such as 32 P-labeled DNA, using filter tips helps prevent the barrel of the micropipettor from getting contaminated.

Hold the micropipettor in a vertical position.

Keeping the micropipettor upright will prevent liquids from running inside and contaminating the barrel of the micropipettor.

The micropipettor has three positions: (1) Rest position, (2) First stop, and (3) Second stop (Figure 6, panel B). The instrument has a two-stop plunger system. The first stop has two functions. The first is to draw in the desired volume of liquid into the tip when releasing the plunger from the first stop to the rest position. The second function is to dispense the majority of liquid from the tip when depressing the plunger from the rest position to the first stop. Further depressing the plunger to the second stop dispenses whatever liquid remains in the tip. Depress the pushbutton on the plunger from the rest position to the first stop. Air equal to the volume of the setting will be displaced.

Immerse the tip into the liquid while holding down the pushbutton to the first stop.

Do not touch the micropipettor itself to the sides of bottles, tubes and flasks otherwise the inside surfaces of these vessels will become contaminated. Only the tips are sterile.

Release the pushbutton slowly to aspirate the liquid into the tip. Stop once the pushbutton is back to the rest position. Wait a moment so liquid can be drawn into the tip.

The volume of liquid in the tip will equal the volume of the setting of the micropipettor.

Viscous liquids such as those containing glycerol require more time to enter the tip.

Remove the tip from the liquid, and visually inspect the tip to confirm that the liquid drawn up has reached the expected level in the tip and there are no air bubbles in the tip.

If necessary, expel the liquid and manually tighten the tips onto the micropipettor. Draw up the liquid and check again.

Place the tip at an angle (10 ° to 45 °) against the wall of the tube receiving the liquid. To expel the liquid, slowly depress the pushbutton on the plunger to the first stop. Wait a moment then press the pushbutton to the second stop to expel any residual liquid in the tip.

Depressing the plunger too quickly may cause the liquid being expelled to splatter or will produce undesirable bubbles in the tube.

Before releasing the plunger to the rest position, remove the tip from the liquid.

Discard tips into designated sharps waste container by pressing the ejection button on the micropipettor.

4. Cleaning Up The Work Space

When finished with an experiment requiring use of aseptic technique, turn off the Bunsen burner, then put away all supplies and reagents. Wipe down the outside surfaces of labware (bottles, micropipettors, pipette tip boxes) with a pre-moistened disinfectant wipe to ensure contaminants are not transferred to the storage location.

Place contaminated glassware and hazardous waste materials into the proper disposal receptacle. Laboratory waste includes labware such as gloves, pipettes, tips, and tubes. Non-infectious hazardous waste is generated when performing experiments with non-pathogenic organisms (BSL-1) while infectious hazardous waste is generated when using pathogenic organisms (BSL-2 or above). Infectious waste must be autoclaved or disinfected before it is discarded. Follow laboratory safety guidelines described in BMBL (5 th Ed.) as well as those provided by OSHA and institutional Environmental Health and Safety departments.

Wipe down the entire work area on the laboratory bench with a pre-moistened disinfectant wipe from the canister, once again allowing the disinfectant to evaporate.

Wash hands thoroughly with antiseptic soap and warm water before leaving the laboratory.

5. Representative Results

A sample application for using serological pipettes to transfer liquids is shown in Figure 7. These pipettes often are used in the microbiology laboratory to prepare media for inoculation with bacterial cultures. For example, sterile flasks first are filled with a specified volume of culture broth, in this case Luria Broth (LB), then a small number of cells (such as E. coli) are added to the media. Using a serological pipette, first the broth must be aseptically transferred from the media bottle to the flask. In this case, 25 ml of LB was added to a 125 ml sterile flask using a 25 ml serological pipette. Next, the broth must be inoculated with E. coli cells. Here, 10 μl of cells were transferred aseptically using a P20 micropipettor from a previously growing culture flask to the 25 ml of fresh LB. The flask is incubated in a growth chamber for a particular amount of time, allowing the cells to replicate (for this example, the E. coli cells were incubated overnight at 37 ଌ on a shaking platform). The result is a turbid bacterial cell culture that can be used for subsequent experiments.

Serological pipettes also may be used to transfer media originally supplied in a bottle to test tubes, or between test tubes, as is done when making dilutions of a bacterial culture. If aseptic technique is not maintained throughout these types of media manipulations, then cultures will become contaminated, and subsequent experiments using those cultures will be delayed because fresh, uncontaminated cultures will need to be prepared. Errors occur because a sterile field is not maintained throughout the procedure. For instance, you may forget to disinfect the laboratory bench or flame the rim of a culture bottle or tube. You may touch the tip of the pipette or set the cap of a bottle or test tube on the bench instead of holding it in your hand. Proper procedure is critical for keeping contamination of media and cultures to a minimum. Figure 8A provides an example of a pure versus contaminated culture of E. coli in a tube containing 5 ml of LB. The left panel shows a culture displaying uniform fine turbidity typical of a pure E. coli culture. In contrast, the right panel shows a contaminated culture in which the growth characteristics deviate from those expected for this bacterial strain.

Technical errors may occur when manipulating serological pipettes resulting in transfer of incorrect volumes of media between test tubes. For instance, you may read the volume on the pipette incorrectly (i.e., top versus bottom of the meniscus) or you may expel the media completely from a TD pipette, which was designed to leave a tiny bit in the tip not to be delivered. When performing a point-to-point delivery of media, you may use the wrong calibration marks and dispense the incorrect volume. Figure 8B shows an example of test tubes with correct versus incorrect volumes of media. The tube on the left contains 3.5 ml of LB measured with a 5 ml serological pipette. The student conducted a point-to-point delivery of the media in which LB was drawn up to the 5.0 ml graduation mark and dispensed to the 1.5 ml mark. The tube on the right contains 2.5 ml of LB measured with a pipette of the same size because the student who performed the point-to-point delivery of media incorrectly dispensed it from the 5.0 ml mark to the 2.5 ml mark. This mistake will result in a bacterial culture that will be at a higher concentration than planned, causing subsequent dilutions to be incorrect. This propagation of errors can result in a failed experiment that would need to be repeated with the correct cell concentrations.

A sample application for using micropipettors to transfer liquids is shown in Figure 9. These pipettors are used for a variety of experiments in molecular biology and microbiology including preparing samples for PCR and gel electrophoresis or inoculating sterile media or buffer with small volumes (less than 1.0 ml) of bacterial cells or phage particles. In the example provided, the student transferred 12.5 μl of TE buffer into a 1.8 ml microcentrifuge tube (left tube in panel A note that dye has been added to the buffer to facilitate visualization of the liquid inside the clear microcentrifuge tubes). This procedure required the student first to select the correct micropipettor, in this case a P20, and next to set the volumeter to the correct volume (panel B). A tip was used that contains a cotton wool plug at the end to prevent possible contamination that could be expelled from the barrel of the micropipettor from reaching the buffer sample in the tip. This precaution is not necessary if care is taken when aspirating liquids into the tips, depressing the plunger slowly so the liquid does not splash into the pipettor barrel. Technical errors may occur that result in transfer of incorrect volumes. For example, you may select the wrong micropipettor for the job or set the volumeter on the correct micropipettor to an incorrect volume. Before immersing the tip into the buffer, you may push the plunger past the first stop, causing an excess of buffer to be drawn into the tip when releasing the plunger. Alternatively, you may not immerse the tip far enough into the buffer, so air is drawn into the tip instead of buffer. You may forget to push the plunger to the second stop when dispensing buffer into the microcentrifuge tube causing less than the desired volume to be released from the tip. The right tube in panel A of Figure 9 shows a microcentrifuge tube containing the incorrect volume of buffer relative to the tube on the left. Instead of dispensing 12.5 μl of buffer, the student dispensed 125 μl. In this case, although the numbers are set identically on the volumeter, the wrong micropipettor was selected for the job (the student used a P200 instead of a P20 panel B) resulting in the delivery of a substantially larger volume of buffer. If this solution was being used to prepare a mixture of reagents for an application such as PCR, then this mistake will change the final concentration of all reagents subsequently added to the same tube. Consequently, it is unlikely that the experiment will be successful, since molecular biology procedures such as PCR require all components to be at specific concentrations for the reaction to work properly.

Because it is not always feasible to ensure micropipettors (especially the inside of the barrel) are sterile, stock solutions can become contaminated causing even troubleshooting efforts to fail when performing experiments. If using micropipettors to transfer sterile solutions, it is strongly recommended that aliquots of stock solutions (media, buffer, water) be made using aseptic technique with serological pipettes. It is common to maintain working stock solutions in 15 ml or 50 ml sterile conical tubes. These are often easier to manipulate while operating a micropipettor and can be replaced with a fresh aliquot of the stock solution if contaminated during volume transfers.

Figure 1. Sterile field created by updraft of Bunsen burner flame. To minimize contamination of sterile solutions and cultures, it is critical that all manipulations be conducted within the sterile field. The rims of glass culture tubes and flasks should be passed through the tip of the blue cone, the hottest part of the flame. Plastic tubes and tips cannot be flamed - these should be pre-sterilized by alternative methods prior to use.

Figure 2. Serological pipettes used for aseptic transfer of liquids. (A) Shown from left to right are drawings of 25 ml, 10 ml, and 5 ml pipettes. (B) Serological pipettes may be plastic or glass. Plastic pipettes are disposable (one-time use) and typically are individually wrapped in paper and plastic sleeves in which all inside surfaces are sterile (left side). Glass pipettes can be used multiple times provided they are cleaned and sterilized between uses these typically are stored in metal canisters (right side).

Figure 3. Serological pipettes are of two types: TC ("to contain") or TD ("to deliver"). Shown is the explanatory label of a TD 5 ml pipette.

Figure 4. Aseptic technique. When aspirating liquids from a bottle, flask, or tube with caps, never place the cap on the bench. Instead, hold the cap in the same hand as pipette aid while manipulating the vessel containing the liquid with the opposite hand as shown.

Figure 5. Meniscus formed when drawing liquid into serological pipette. The volume corresponds to the graduation mark on the pipette where the bottom of the meniscus aligns. In this example, the meniscus aligns with the 2.5 ml graduation mark.

Figure 6. Single channel micropipettor. (A) Shown is a sample micropipettor with a plastic tip attached to the bottom of the barrel tip holder. Indicated are the locations of the volumeter, the thumb wheel for changing the volumeter setting, the barrel tip holder, the tip ejector button, and the pushbutton for the plunger. (B) Two-stop plunger system on a micropipettor.

Figure 7. Using serological pipettes to transfer media into sterile 125 ml flasks. The left flask has 25 ml of media only (LB), while the right flask is a culture of E. coli resulting from inoculating LB with cells then incubating overnight at 37 ଌ. Note how the media in the flask on the right is turbid due to cell growth.

Figure 8. Using serological pipettes to transfer media into sterile test tubes. (A) The left tube contains 5 ml of a pure E. coli culture, while the right tube contains 5 ml of a contaminated bacterial cell culture. Note the differences in growth characteristics between the two cultures. Although both are turbid, the culture on the right has been contaminated with a fungus or other airborne microorganisms giving the culture a different color and consistency from that expected for E. coli cells. (B) The left culture tube contains 3.5 ml LB while the right tube contains only 2.5 ml LB. This volume difference resulted from a mistake made while conducting a point-to-point delivery of media to the tubes.

Figure 9. Using micropipettors to transfer buffer into sterile microcentrifuge tubes. (A) The left microcentrifuge tube contains only 12.5 μl of TE buffer, while the right tube contains 125 μl. Note that a dye has been added to the buffer to facilitate visualization of the liquid inside the clear microcentrifuge tubes. (B) The left volumeter is from a P20 micropipettor, while the right volumeter is from a P200 micropipettor. A common mistake is selecting the wrong micropipettor. Although the numbers are set identically on the P20 and P200 volumeter, selection of the wrong micropipettor results in transfer of incorrect volumes.

Figure 10. Laminar flow hood used to prevent contamination of solutions and cultures. Shown is a biosafety cabinet approved for work with BSL-2 organisms.


Biology

BIOL 1300 Body Systems with Lab (3 semester credit hours) Examines the organ systems of mammals, predominantly the human. Function in relation to structure is emphasized. The effects of one organ system on others are stressed. The overall objective of the course is an appreciation of the integration and control of all systems. There is a model-based human anatomy lab. This course is specifically designed for non-majors. (2-2) S

BIOL 1318 (BIOL 2316) Human Genetics (3 semester credit hours) Elementary course in the fundamentals of human genetics. Topics include patterns of inheritance DNA structure and replication gene function mutation and its role in genetic diseases, cancer, and the immune system matters of sex evolution genetic engineering and gene therapy forensics and bioethics. This course is specifically designed for non-majors. (3-0) Y

BIOL 1350 Body Systems (3 semester credit hours) Examines the organ systems of mammals, predominantly the human. Function in relation to structure is emphasized. The effects of one organ system on others is stressed. The overall objective of the course is an appreciation of the integration and control of all systems. This course is specifically designed for non-majors. (3-0) R

BIOL 1V00 Topics in Biological Sciences (1-6 semester credit hours) May be repeated for credit as topics vary (6 semester credit hours maximum). ([1-6]-0) R

BIOL 1V01 Topics in Biological Sciences with Lab (1-6 semester credit hours) May be repeated as topics vary (6 semester credit hours maximum). ([1-5]-[1-5]) R

BIOL 1V95 Individual Instruction in Biology (1-6 semester credit hours) Individual study under a faculty member's direction. May be repeated for credit as topics vary (6 semester credit hours maximum). Instructor consent required. ([1-6]-0) S

BIOL 2111 Introduction to Modern Biology Workshop I (1 semester credit hour) Problem solving and discussion related to the subject matter in BIOL 2311. Prerequisites: ((CHEM 1311 or CHEM 1315 or equivalent) and (CHEM 1312 or CHEM 1316)) or CHEM 1301. Corequisite: BIOL 2311. (1-0) S

BIOL 2112 Introduction to Modern Biology Workshop II (1 semester credit hour) Problem solving and discussion related to the subject matter in BIOL 2312. Corequisite: BIOL 2312. (1-0) S

BIOL 2281 Introductory Biology Laboratory (2 semester credit hours) Introductory lectures discuss the theoretical and historical aspects of the experiments carried out in the laboratory. Laboratory experiments introduce the student to bioinformatics, basic cellular biology, and structure and function of proteins and nucleic acids. Computer exercises in bioinformatics involve multiple alignment analyses, BLAST and literature searches, and construction of phylogenetic trees. Laboratory experiments include microscopy, microbial techniques, yeast genetics, and the electrophoretic behavior of normal and mutant proteins. DNA related experiments include isolation (nuclear and mtDNA), amplification, restriction digests, electrophoresis, plasmid mapping, and transformations. Students present posters of their long-term investigations at the end of the semester. Prerequisite: BIOL 2311 (also see prerequisites for BIOL 2311). ([0-1]-[1-2]) S

BIOL 2311 (BIOL 1306) Introduction to Modern Biology I (3 semester credit hours) Presentation of some of the fundamental concepts of modern biology, with an emphasis on the molecular and cellular basis of biological phenomena. Topics include the chemistry and metabolism of biological molecules, elementary classical and molecular genetics, and selected aspects of developmental biology, physiology (including hormone action), immunity, and neurophysiology. Prerequisites: ((CHEM 1311 or CHEM 1315) and (CHEM 1312 or CHEM 1316)) or CHEM 1301. Corequisite: BIOL 2111. (3-0) S

BIOL 2312 (BIOL 1307) Introduction to Modern Biology II (3 semester credit hours) The overall emphasis will be on organ physiology and regulatory mechanisms involving individual organs and organ systems. Factors considered will be organ development and structure, evolutionary processes and biological diversity, and their effects on physiological mechanisms regulating the internal environment. Corequisite: BIOL 2112. (3-0) S

BIOL 2350 Biological Basis of Health and Disease (3 semester credit hours) Fundamentals of pathophysiology, focusing on the dynamic processes that cause disease, give rise to symptoms, and signal the body's attempt to overcome disease. The course covers diseases which may affect dramatically the life of an individual and society in the modern age. Topics include 1) mechanisms of infectious disease, immunity, and inflammation and 2) alterations in structure and function of the reproductive, circulatory, respiratory, and urinary systems. Special emphasis is given to preventative aspects for each disease based on non-drug, wellness-promoting approaches. This course is designed as a science elective open to all majors. (3-0) S

BIOL 2V00 Topics in Biological Sciences (1-6 semester credit hours) May be repeated as topics vary (6 semester credit hours maximum). Instructor consent required. ([1-6]-0) R

BIOL 2V01 Topics in Biological Sciences with Lab (1-6 semester credit hours) May be repeated as topics vary (6 semester credit hours maximum). ([1-5]-[1-5]) R

BIOL 2V95 Individual Instruction in Biology (1-6 semester credit hours) Individual study under a faculty member's direction. May be repeated for credit as topics vary (6 semester credit hours maximum). Instructor consent required. ([1-6]-0) S

BIOL 3101 Classical and Molecular Genetics Workshop (1 semester credit hour) Problem solving and discussion related to the subject matter in BIOL 3301. Prerequisites: BIOL 2311 and (BIOL 2281 or CHEM 2401 or equivalent) and (CHEM 2323 or equivalent). Corequisite: BIOL 3301. (1-0) S

BIOL 3102 Eukaryotic Molecular and Cell Biology Workshop (1 semester credit hour) Problem solving and discussion related to the subject matter in BIOL 3302. Prerequisites: BIOL 3301 and (BIOL 3361 or CHEM 3361) or equivalent. Corequisite: BIOL 3302. (1-0) S

BIOL 3161 Biochemistry Workshop I (1 semester credit hour) Problem solving methodology in biochemistry discussion of recent advances in areas related to the subject matter in BIOL 3361 or CHEM 3361. Prerequisites: (CHEM 2323 or equivalent) and CHEM 2325. Corequisite: BIOL 3361 or CHEM 3361. (1-0) S

BIOL 3162 Biochemistry Workshop II (1 semester credit hour) Problem-solving methodology in biochemistry discussion of recent advances in areas related to the subject matter in BIOL 3362 or CHEM 3362. Prerequisite: BIOL 3361 or CHEM 3361 or equivalent, or instructor consent required. Corequisite: BIOL 3362 or CHEM 3362. (1-0) Y

BIOL 3301 Classical and Molecular Genetics (3 semester credit hours) The phenomenon of heredity, its cytological and molecular basis gene expression and transfer of genetic information, with major focus on bacterial and model eukaryotic systems genetic recombination and chromosome mapping tetrad analysis mutations and mutagenesis genetic interactions application of recombinant DNA techniques to genetic analysis. Prerequisites: BIOL 2311 and (BIOL 2281 or CHEM 2401 or equivalent) and (CHEM 2323 or equivalent). Corequisite: BIOL 3101. (3-0) S

BIOL 3302 Eukaryotic Molecular and Cell Biology (3 semester credit hours) Structural organization of eukaryotic cells regulation of cellular activities membranes and transport cellular replication examples of cell specialization such as blood (immunoglobulins) and muscle cells. Prerequisites: BIOL 3301 and (BIOL 3361 or CHEM 3361) or equivalent. Corequisite: BIOL 3102. (3-0) S

BIOL 3303 Introduction to Microbiology (3 semester credit hours) Microbes contribute to major biogeochemical processes, live in environments inhospitable to other organisms, and may comprise the majority of biomass on Earth. They form beneficial symbioses with multicellular organisms and play critical roles in the development of those organisms. In contrast to these beneficial roles, certain microbes are global public health concerns. This course surveys the form and function of the microbial world. Prerequisites: (BIOL 2281 or equivalent) and BIOL 2311 and BIOL 2312. (3-0) S

BIOL 3305 Evolutionary Analysis (3 semester credit hours) Molecular and fossil evidence for evolution. Darwinian natural selection, mechanisms of evolution, Mendelian genetics in populations, forms of adaptation, evolutionary trees, molecular phylogeny, theories on the origin of life. Prerequisite: BIOL 3301. (3-0) Y

BIOL 3312 Introduction to Programming for Biological Sciences (3 semester credit hours) This course is an introduction to programming practices using C++ designed specifically for students in the biological sciences. Special emphasis will be put in particular features of C++ like object oriented programming, some data structures as well as applications to process, model and analyze biological data. One goal of this course is to provide a strong background on programming skills on a basic level while leaving more advanced techniques of software development and algorithms for other advanced courses. This course also covers an introduction to data analysis with R, a statistical platform used widely in the biological sciences community. Prerequisites: (BIOL 2281 or equivalent) and BIOL 2311 and BIOL 2312. (3-0) Y

BIOL 3315 Epigenetics (3 semester credit hours) Almost all cell types in our body share the same genetic information, but they perform very different functions. For example, our nerve cells are morphologically and functionally distinct from our muscle cells. How can the same genome give rise to hundreds of distinct cell types in our body? How can different diseases affect identical twins sharing the same genetic information? Why our parents and grandparents diet and health may have lasting influences in our own health? The field of epigenetics emerged over the past decades to tackle these fundamental questions that intersect our genome, development, environment and disease. This course will provide a broad overview of epigenetic phenomena and epigenetic mechanisms with weekly lectures and small group discussion of primary literature. The course will introduce students to seminal works in epigenetics and recent developments with the goal of instilling critical knowledge of the field. Prerequisites: (BIOL 3101 and BIOL 3301) or equivalent or instructor consent required. (3-0) Y

BIOL 3318 Forensic Biology (3 semester credit hours) Role and methodology of biological testing in criminal investigation and forensic science. Analysis of the procedures and methodologies employed in the collection, preservation and screening of biological evidence, and protein and DNA testing. Population genetics employed during the statistical evaluation of data is covered. The course is structured to allow individuals with and without biological training to participate. The subject matter will be developed from the concept of "What is DNA?" through "What does a statistical estimate really mean?" (3-0) T

BIOL 3320 Applied Genetics (3 semester credit hours) Genetic model organisms such as the flatworm (Planaria), fruit fly (Drosophila melanogaster), nematode (Caenorhabditis elegans), and the zebrafish (Danio rerio) are the cornerstones of biomedical research. These organisms known for their simplicity of structure and gene similarity to humans have been seminal in advancing our understanding of many biological processes and human diseases. In this inquiry-based course, learners will apply basic principles of genetic model systems, transmission genetics, and molecular genetics to investigate important biological concepts such as embryonic cell division, stem cells and regeneration, Mendelian inheritance, gene mutations, and phenotypes. Throughout this exploratory course, students will gain practical hands-on experience conducting basic culturing, genetic manipulation and phenotypic analysis necessary to utilize genetic model organisms in their investigation. Learners will engage in class discussions and activities to draw connections between the concepts learned in class and their real-time application(s) in biomedical sciences. Prerequisites: BIOL 2281 or equivalent and (BIOL 2311 and (BIOL 2111 or equivalent)) and (BIOL 2312 and (BIOL 2112 or equivalent)). (3-0) S

BIOL 3335 Microbial Physiology (3 semester credit hours) Life processes of microbes: fermentations, N2 assimilation, and other biochemical pathways specific to bacteria cellular structure and differentiation, among others. Substitutes for BIOL 3362 or CHEM 3362 for Biology majors. Prerequisites: BIOL 2311 and (BIOL 3361 or CHEM 3361). (3-0) T

BIOL 3336 Protein and Nucleic Acid Structure (3 semester credit hours) Examines the different types of protein motifs, protein and DNA folding and stability, and the relation of structure to function. Circular dichroism, NMR, and crystallographic methods of structural determination are presented. Types of proteins considered include transcription factors, proteinases, membrane proteins, proteins in signal transduction, proteins of the immune system, and engineered proteins. Students also receive instruction in the viewing and manipulation of protein and DNA structures using various modeling programs and data from national web sites. Prerequisite: BIOL 3361 or CHEM 3361. (3-0) T

BIOL 3351 Medical Cell Biology (3 semester credit hours) Explores topics in cell biology and medicine. Topics include cellular organization, structure and inheritance of DNA, gene therapy, stem cells, regenerative medicine, cell to cell signaling, the functioning of different types of cells and tissues, including those of the immune and endocrine system, and the study of several genetic diseases, such as cancer and cardiovascular disease. Prerequisites: BIOL 2311 and BIOL 2312 or equivalent. (3-0) S

BIOL 3355 Clinical Pathophysiology (3 semester credit hours) The focus of this course is to meet the interests of the students who plan to become professionals working in the health-care field. The strategic goal of the course is to make students internalize the notion of the complexity of the processes leading to the onset and the development (pathogenesis) of a diseased condition, to emphasize the concept of the unbalanced homeostatic regulation underlying any pathology. To understand the idea of the involvement of all body systems in the seemingly "local" manifestations of a disease, and to realize the importance of the mind-body connections in the subjective and objective characteristics of an individual ailment and its influence on the process of sanogenesis (recovery). We will incorporate the most recent scientific data into the fundamentals of pathophysiology and discuss the classical typological problems like the etiology, diagnosis, clinical characteristics, treatment, and the prognosis of the condition. The pathological conditions that will be covered in this course include the infectious diseases and some immune disorders, the diseases of the reproductive, cardiovascular, respiratory, and urinary systems. Prerequisites: BIOL 2281 and BIOL 2312. ([1-3]-0) S

BIOL 3357 Mammalian Physiology with Lab (3 semester credit hours) This course will focus on human body systems and physiological pathways related to organ system functions and control including, but not limited to, central nervous system control and feedback, cardiovascular, respiratory, and neuromuscular physiology as well as topics such as blood pressure regulation and exercise physiology. This course will use computer software and electronic instrumentation for performing electrocardiography, electromyography, electroencephalography, plethysmography, pulmonary function analysis, polygraph analysis, and biofeedback. Instructor consent required. Prerequisites: BIOL 3455 or equivalent and BIOL 3456 or equivalent. (3-1) S

BIOL 3361 Biochemistry I (3 semester credit hours) Structures and chemical properties of amino acids protein purification and characterization protein structure and thermodynamics of polypeptide chain folding catalytic mechanisms, kinetics and regulation of enzymes energetics of biochemical reactions generation and storage of metabolic energy associated with carbohydrates oxidative phosphorylation and electron transport mechanisms photosynthesis. Prerequisites: CHEM 2323 (or equivalent) and CHEM 2325 (or equivalent). Corequisite: BIOL 3161. (Same as CHEM 3361) (3-0) S

BIOL 3362 Biochemistry II (3 semester credit hours) Breakdown and synthesis of lipids membrane structure and function nitrogen metabolism and fixation nucleotide metabolism structure and properties of nucleic acids sequencing and genetic engineering replication, transcription, and translation chromosome structure hormone action biochemical basis of certain pathological processes. Prerequisite: (BIOL 3361 or CHEM 3361) or its equivalent, or instructor consent required. Corequisite: BIOL 3162. (Same as CHEM 3362) (3-0) S

BIOL 3370 Exercise Physiology (3 semester credit hours) Examines the operation and adaptation of human organ systems (cardiovascular, respiratory, renal, skeletal, and hormonal) during exercise. Clinical aspects of exercise, including the effects of training, nutrition, performance, and ergogenic aids, are also discussed. Prerequisites: BIOL 2312 and (BIOL 3455 or BIOL 3456 or equivalent). (3-0) Y

BIOL 3380 Biochemistry Laboratory (3 semester credit hours) Current techniques in the purification and characterization of enzymes to demonstrate fundamental principles that are utilized in modern biochemistry and molecular biology research laboratories. Practical skills taught include micropipetting, basic solution preparation, conducting pH measurements, isolating crude enzyme extracts, and performing standard activity assays. Advanced experiments with Green Fluorescent Protein and Lactate Dehydrogenase include Ni++-NTA affinity chromatography, ion chromatography, protein detection using Bradford, Lowry, and spectrophotometric assays, SDS-PAGE separation, Western Blot analysis, and enzyme kinetics. Prerequisite: BIOL 2281 or CHEM 2401 or equivalent. Prerequisite or Corequisite: BIOL 3361 or CHEM 3361. (1-4) S

BIOL 3385 Medical Histology (3 semester credit hours) Medical histology will cover the microscopic structure and function of human cells and tissues that make up the organ systems in normal and pathological conditions. The lecture component will include understanding of relevant disease and pathophysiological conditions from a histological standpoint. The laboratory component of this course will involve the microscopic study of cells and tissues using the compound light microscope and prepared slides. Laboratory studies will complement and correlate with the study of cells and tissue organization. Prerequisites: BIOL 2311 and BIOL 2312. (1.5-3) S

BIOL 3388 Honey Bee Biology (3 semester credit hours) This survey course explores the biology of honey bees at the colony, organism, and molecular levels. Topics include honey bee anatomy, nest architecture, caste development and social organization, reproduction and genetic diversity, pheromones and communication, foraging behavior, colony reproduction, pest and disease management, and basic beekeeping. Optional hands on experience may be provided. Prerequisites: (BIOL 2281 or CHEM 2401 or equivalent) and BIOL 2311 and BIOL 2312. (3-0) Y

BIOL 3455 Human Anatomy and Physiology with Lab I (4 semester credit hours) First of a two-course sequence providing a comprehensive study of the basic principles of human physiology in conjunction with a detailed, model-based human anatomy laboratory and computer-assisted physiology experiments. Examination of structure-function relationships includes a survey of human histology and skeletal, muscular, neural, and sensory organ systems. Prerequisite: BIOL 2312 or equivalent. (3-3) S

BIOL 3456 Human Anatomy and Physiology with Lab II (4 semester credit hours) Continuation of the comprehensive study of the basic principles of human physiology in conjunction with a detailed, model-based human anatomy laboratory and computer-assisted physiology experiments. Endocrine, cardiovascular, respiratory, digestive, renal, and reproductive systems are examined. Prerequisite: BIOL 3455 or equivalent. (3-3) S

BIOL 3520 General Microbiology with Lab (5 semester credit hours) Majors course in general microbiology. Lectures include topics recommended by the Education Division of the American Society for Microbiology: microbial structure, diversity, growth and growth control, metabolism, genetics, and gene regulation. Among additional topics covered are virology, immunology and microbial diseases (plant and animal) including epidemiology, transmission, and host-microbe interactions. The laboratory focuses on developing laboratory skills in classical microbiology by the individual student. Exercises include various staining and pure culture techniques, biochemical and other in vitro testing, as well as isolation and identification of unknown organisms. Prerequisites: (BIOL 2281 or CHEM 2401 or equivalent) and (BIOL 2311 and BIOL 2312) or equivalent and CHEM 2323. (2-3) Y

BIOL 3V00 Topics in Biological Sciences (1-6 semester credit hours) May be repeated as topics vary (9 semester credit hours maximum). Prerequisites: (BIOL 2281 or CHEM 2401 or equivalent) and BIOL 2311 and BIOL 2312 or equivalent. ([1-6]-0) S

BIOL 3V01 Topics in Biological Sciences with Lab (1-6 semester credit hours) May be repeated as topics vary (6 semester credit hours maximum). Prerequisites: (BIOL 2281 or CHEM 2401 or equivalent) and BIOL 2311 and BIOL 2312 or equivalent. ([1-5]-[1-5]) R

BIOL 3V15 Research Practicum for UT-PACT (1-6 semester credit hours) Students in the UT-PACT program participate in clinical or biomedical research projects under the joint supervision of UT Southwestern faculty and UT Dallas UT-PACT program coordinator. Students receive training in relevant research methodology and research ethics prior to placement in clinical settings. Consult with UT-PACT program coordinator prior to enrollment for information on prerequisites and minimum on-site hours. May be repeated for credit. (9 semester credit hours maximum). UT-PACT program coordinator consent required. ([1-6]-0) S

BIOL 3V40 Topics in Molecular and Cell Biology (1-6 semester credit hours) May be repeated as topics vary (9 semester credit hours maximum). Prerequisites: (BIOL 2281 or CHEM 2401 or equivalent) and BIOL 2311 and BIOL 2312 or equivalent. ([1-6]-[0-5]) S

BIOL 3V81 Clinical Medicine I (1-6 semester credit hours) Clinical Medicine is a component of the UT Partnership in Advancing Clinical Transition (UT PACT) program that addresses clinical competencies in the medical profession, including communication skills, professional identity formation, interprofessional teamwork, and medical ethics. Students participate in small group sessions, clinical preceptorships, and hospital rotations at UT Southwestern Medical Center. Enrollment is limited to students who have completed at least one year of the UT PACT Program. Credit/No Credit only. UT PACT advisor consent required. ([1-6]-[1-9]) Y

BIOL 3V82 Clinical Medicine II (1-6 semester credit hours) Clinical Medicine II addresses clinical competencies in the medical profession, building on skills already addressed in Clinical Medicine I and other parts of the UT Partnership in Advancing Clinical Transition (UT PACT) curriculum. Topics to be addressed include the application of basic science to clinical practice, interpersonal skills in medicine, cultural competency, and professionalism and medical ethics in clinical settings. Students participate in small group sessions and clinical preceptorships and rotations at UT Southwestern Medical Center. Enrollment is limited to students who have completed their second year in the UT PACT Program. Credit/No Credit only. UT PACT advisor consent required. Prerequisite: BIOL 3V81. ([1-6]-[1-9]) Y

BIOL 3V83 Clinical Medicine III (1-6 semester credit hours) Clinical Medicine III is a continuation of Clinical Medicine I and II that is offered to students in the UT Partnership in Advancing Clinical Transition (UT PACT) program, to be taken during students' third academic year at UT Dallas. Enrollment is limited to students who have completed Clinical Medicine I and II, and at least two years of the UT PACT Program. UT PACT advisor consent required. ([1-6]-[1-9]) Y

BIOL 3V84 Clinical Medicine IV (1-6 semester credit hours) Clinical Medicine IV is a continuation of Clinical Medicine I, II, and III that is offered to students in the UT Partnership in Advancing Clinical Transition (UT PACT) program to be taken during students' third academic year at UT Dallas. Enrollment is limited to students who have completed Clinical Medicine I, II, and III, and at least two years of the UT PACT Program. Credit/No Credit only. UT PACT advisor consent required. ([1-6]-[1-9]) Y

BIOL 3V90 Undergraduate Readings in Biology (1-3 semester credit hours) Subject and scope to be determined on an individual basis. May be repeated for credit as topics vary. Instructor consent required. ([1-3]-0) S

BIOL 3V91 Undergraduate Research in Biology (1-3 semester credit hours) Subject and scope to be determined on an individual basis. May be repeated for credit as topics vary. Instructor consent required. ([1-3]-0) S

BIOL 3V93 Undergraduate Research in Biochemistry (1-3 semester credit hours) Subject and scope to be determined on an individual basis. May be repeated for credit as topics vary. Instructor consent required. ([1-3]-0) S

BIOL 3V94 Topics in Biology: Individual Instruction (1-6 semester credit hours) Individual study under a faculty member's direction. May be repeated for credit as topics vary. Instructor consent required. ([1-6]-0) S

BIOL 3V96 Undergraduate Research in Molecular and Cell Biology (1-3 semester credit hours) Subject and scope to be determined on an individual basis. May be repeated for credit as topics vary. Instructor consent required. ([1-3]-0) S

BIOL 4302 TA Apprenticeship (3 semester credit hours) Development and practice of teaching skills in the classroom and laboratory in the biological sciences. May be repeated only once for credit (6 semester credit hours maximum). Instructor consent required. (3-0) S

BIOL 4305 Molecular Evolution (3 semester credit hours) This course describes principles and models of evolutionary theory at the molecular level. It focuses primarily on the evolution of nucleotide sequences including genes, pseudogenes, and genomes as well as amino acid sequences used to study the evolution of proteins, protein complexes, and interactions. Phylogenetics and current leading quantitative models of sequence evolution are discussed in detail. Recent methods on amino acid evolution and its connections to molecular structure and function are also studied. Relevant examples of molecular evolution presented in this course include protein interactions, signaling networks, and viral evolution. Students learn computational tools and algorithms used to study evolution at the molecular level and work on a proposal-like research project applying tools and concepts learned in class to investigate new research questions in their area of specialization. Prerequisites: BIOL 3301 and BIOL 3302. (3-0) S

BIOL 4310 Cellular Microbiology (3 semester credit hours) The course covers topics related to pathogenesis of infectious diseases in the context of host cell properties. It introduces various human pathogens and describes their virulence, and explores the evolutionary aspects of how pathogens interact with their host cells and how host cells defend themselves against invading microorganisms. Topics include bacterial toxins and secretion mechanisms, virus infections, microbial invasion and intracellular parasitism, manipulation of host cell functions and induction of cell death by pathogens, innate and acquired defense mechanisms of the host, inflammation, sepsis, and advances of microbial genomics involving human microbiome, vaccines, and anti-infectives. The course aims to complement the scientific knowledge and principles established in cell biology, medical microbiology, and immunology with appropriate relevance to clinical applications involving parasitology and infectious disease control. Prerequisite: BIOL 2311. (3-0) Y

BIOL 4315 Genes, Disease and Therapeutics (3 semester credit hours) This course explores models of genetic disease beginning with the genetic basis and traveling through the clinical presentation. Therapeutic approaches as well as particular issues relevant to each disease are also covered. These issues include legal aspects, prenatal screening and ethical concerns. Prerequisites: BIOL 2311 and BIOL 2312 and (BIOL 2281 or CHEM 2401 or equivalent). (3-0) S

BIOL 4317 Cellular and Molecular Medicine of Human Diseases (3 semester credit hours) This course is designed to provide upper level undergraduate students with current understandings of and experimental approaches (e.g. animal models) to human diseases with emphasis on cellular and molecular basis of cancer, metabolic diseases, inflammation, and tissue injuries. Students will become aware of the most recent advancements in biomedical research and the contributions of various animal models to basic and clinical studies. Students are also expected to acquire the necessary skills to interpret and present recent landmark research articles. Sessions include lectures, seminars from invited guest lecturers, and journal article presentation. Prerequisites: (BIOL 3301 and BIOL 3302 and BIOL 3361) or instructor consent required. (3-0) R

BIOL 4320 Cell Migration in Health and Disease (3 semester credit hours) Cell adhesion and migration play important roles in normal development, immune responses, wound healing and regeneration. Dysregulated migration underlies many conditions including congenital disorders, chronic inflammation, and cancer invasion and metastasis. This course will examine the cellular and molecular mechanisms underlying cell adhesion and migration in normal, regenerative and diseased states. Model systems, tools, and technologies used to study and analyze cell migration will be discussed. The course will include didactic lectures, enquiry-based learning and student presentations. Prerequisites: (BIOL 3301 and BIOL 3302) and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. (3-0) S

BIOL 4325 Nutrition and Metabolism (3 semester credit hours) This course examines nutrient utilization and requirements with an emphasis on multifaceted links between diet, health, genetics, microbiome, and diseases. The course intends to support studies towards medicine, health professions, biomedical research, and biotechnology. Topics cover the basis of nutritional physiological phenomena and metabolic hemostasis in the context of human development, aging, exercise, health and diseases. Integration of energy metabolism and physiological requirements concerning macronutrients and major vitamins and minerals as well as benefits of potentially-protective compounds in food are reviewed. How unbalanced intake of nutrients contributes to the initiation, development and severity of various chronic diseases, including coronary heart disease, atherosclerosis, lipidemia, hypertension, diabetes, obesity, osteoporosis, thyroid disorders, immune dysfunction, inflammatory conditions, cancer, and dysbiosis are discussed with relevance to clinical nutrition and public health. The course also introduces the fields of microbiomics, nutrigenomics, nutrigenetics and chrononutrition to explore evolving concepts concerning the influence of diet on intestinal microbiota and the effect of foods and sleep on metabolism and genes. Prerequisites: (BIOL 3361 and BIOL 3161) or equivalent and (BIOL 3362 and BIOL 3162) or equivalent. (3-0) S

BIOL 4330 Advanced Research in Molecular and Cell Biology (3 semester credit hours) This course aims to show students how to carry out original research and to teach them some practical approaches and techniques used in a research laboratory. Advanced research approaches and techniques will be used to investigate fundamental molecular and cellular processes in eukaryotic cells and organisms. Practical skills that will be taught and applied include the following: growth and monitoring of bacterial and yeast cultures, plasmid DNA isolation, restriction digest analysis, DNA cloning, polymerase chain reaction (PCR), bacterial and yeast transformation with DNA. Advanced techniques include fluorescent microscopy, beta-galactosidase, and fluorescent reporter assays, cancer cell cultures, protein extraction, protein purification, and immunohistochemistry. Prerequisites: BIOL 2281 and BIOL 2311 and BIOL 3302 and CHEM 2125. (1-5) S

BIOL 4337 Seminal Papers in Biology (3 semester credit hours) Theoretical and experimental papers in selected areas of biology will be discussed in a senior seminar format. The historical and biographical context of the papers and their authors will also be explored. The areas to be covered in any semester will vary with the instructor. Each student is expected to make an oral presentation and to prepare a written paper. Prerequisites: (BIOL 3301 and BIOL 3302) and (BIOL 3361 or CHEM 3361) and (BIOL 3362 or CHEM 3362). (3-0) S

BIOL 4341 Genomics (3 semester credit hours) Fundamentals of how the human genome sequence was acquired and the impact of the human genome era on biomedical research, medical care and genetic testing. Also covered is the impact new tools such as DNA microarray, real time PCR, mass spectrometry and bioinformatics will have on approaches to how scientific questions are investigated. The class will be a mixture of didactic lectures and paper presentations on examples of applied genomics. There will be two computer-based labs where students will perform online bioinformatics and data mining using the NCBI public database. Prerequisite: BIOL 3301 with a grade of C or better. (3-0) T

BIOL 4345 Immunobiology (3 semester credit hours) Interactions of antigens and antibodies. Fine structure of antibodies. Tissues and cells of the immune system. Response of B and T lymphocytes to antigens. Cellular interactions in humoral and cell-mediated immunity. Genetic basis of antibody diversity. Immunity and infectious diseases. Prerequisites: CHEM 2323 and CHEM 2325 (Organic Chemistry I and II). Suggested additional preparation: BIOL 3302. (3-0) T

BIOL 4350 Medical Microbiology (3 semester credit hours) This course will cover the methods used for identification of pathogenic organisms and the study of these organisms in relation to their disease process in humans. We will also cover at the molecular level important concepts such as microbial virulence, the control of bacterial growth, and host responses to infection. Prerequisite: BIOL 3301 or BIOL 3V20. (3-0) T

BIOL 4353 Molecular Biology of HIV/AIDS (3 semester credit hours) Topics include a discussion of the history and epidemiology of AIDS, the likely origins of human immunodeficiency virus (HIV), and the molecular and cell biology of HIV replication. The cell biological basis of the immunodeficiency induced by HIV infection is examined, as well as that of common accompanying pathologies such as Kaposi's sarcoma. The molecular basis of a variety of existing and potential anti-viral therapies is considered. Suggested prerequisite: BIOL 3302. (3-0) T

BIOL 4356 Molecular Neuropathology (3 semester credit hours) Molecular Neuropathology course offers a 360 degree view on neurological diseases and the underlying molecular causes. In this course, we will be looking at the pathology of the brain and CNS in various diseases. Following a look at the pathology, we will dive into the molecular aspects of the same diseases looking at it from the genetic and protein structure-function point of view. We love an open class format and enjoy discussions on the various topics on the syllabus. Prerequisites: BIOL 3301 and BIOL 3302 and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. (3-0) S

BIOL 4357 Molecular Neuropathology II (3 semester credit hours) Molecular Neuropathology course offers a 360 degree view on neurological diseases and the underlying molecular causes. In this course, we will be looking at the pathology of the brain and CNS in various diseases. Following a look at the pathology, we will dive into the molecular aspects of the same diseases looking at it from the genetic and protein structure-function point of view. We love an open class format and enjoy discussions on the various topics on the syllabus. Prerequisites: BIOL 3301 and BIOL 3302 and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. (3-0) Y

BIOL 4360 Evolution and Development (3 semester credit hours) The objective of the course is to integrate evolutionary biology and developmental biology into a common framework, focusing on the evolution of developmental pathways as a basis for the evolution of animal morphology. This is a reading intensive course with a heavy focus on scientific research. Prerequisite or Corequisite: BIOL 3301. (3-0) S

BIOL 4365 Advanced Human Physiology (3 semester credit hours) Function and integration of human organ systems. The role of these systems in the adaptation of humans to, and their interaction with, the environment. Maintenance and perturbation of homeostasis. Pathophysiological basis of certain diseases. Prerequisite: BIOL 3302 or instructor consent required. (3-0) R

BIOL 4366 Molecular Biology of Cancer (3 semester credit hours) Subject matter includes a discussion of representative examples of the principal categories of dominantly acting oncogenes. The role in oncogenesis of tumor suppressor genes ("recessive oncogenes") is also considered, as are anti-apoptotic oncogenes such as Bcl. The roles that the proteins encoded by these genes play in growth hormone signal transduction, gene regulation, cell cycle regulation, and programmed cell death will be examined. Students will also read and discuss the primary literature in this field. Prerequisite: BIOL 3302. (3-0) T

BIOL 4371 General and Molecular Virology (3 semester credit hours) What is a virus? What is the basis of virus/host specificity? How do viruses replicate? This course will cover virus structure, classification, gene expression, and replication. Once we have covered the basics using a few select model systems, we will consider selected groups of viruses from each of the three domains of life and discuss in detail virus replication from attachment to release of progeny virions (and/or alternative fates such as lysogeny, abortive infections and others). This course is designed for upper level undergraduate students who have a firm grasp on the basics of Central Dogma: transcription, translation, replication, as well as a background in bacteriology and eukaryotic cell biology. BIOL 3302 is recommended but not required. Prerequisites: BIOL 3301 and (BIOL 3520 or BIOL 3V20) or instructor consent required. (3-0) Y

BIOL 4380 Cell and Molecular Biology Laboratory (3 semester credit hours) Current techniques that are utilized in a modern molecular biology research laboratory. Practical skills taught include monitoring bacterial growth, phenotype testing, plasmid isolation, restriction digest analysis, DNA cloning, and DNA fingerprinting using the polymerase chain reaction (PCR). Advanced techniques include fundamental microscopy, DNA transfection and general characterization of animal cell cultures, sub-cellular fractionation using differential centrifugation, basic immunological techniques, and chemical mutagen testing. Prerequisite: BIOL 3380. Prerequisite or Corequisite: BIOL 3302. (1-4) S

BIOL 4385 Oral Histology and Embryology (3 semester credit hours) This course will provide exposure to and broad coverage of maxillofacial and oral histological structures and embryology of the face, neck, and teeth using lectures and electronic images of calcified and soft tissues cells. Prerequisites: (BIOL 3361 and (BIOL 3455 or BIOL 3456)) or instructor consent required. (3-0) S

BIOL 4390 Senior Readings in Molecular and Cell Biology (3 semester credit hours) For students conducting independent literature research and scientific writing in Biology or Molecular and Cell Biology. Subject and scope to be determined on an individual basis. Topics may vary. Instructor consent required. (3-0) S

BIOL 4391 Senior Research in Molecular and Cell Biology (3 semester credit hours) For students conducting laboratory research and scientific writing in Biology or Molecular and Cell Biology. Subject and scope to be determined on an individual basis. Topics may vary. Instructor consent required. (3-0) S

BIOL 4399 Senior Honors Research for Thesis in Molecular and Cell Biology (3 semester credit hours) For students conducting independent laboratory research for honors in Biology or Molecular and Cell Biology. Besides the university specifications the student should contact the undergraduate academic advisor in biology for program requirements. Topics may vary. Instructor consent required. (3-0) S

BIOL 4461 Biophysical Chemistry (4 semester credit hours) For students interested in the interface between biochemistry and structural biology. Provides an advanced treatment of the physical principles underlying modern molecular biology techniques. Topics include classical and statistical thermodynamics, biochemical kinetics, transport processes (e.g., diffusion, sedimentation, viscosity), chemical bonding, and spectroscopy. Prerequisites: ((MATH 2413 and MATH 2414) or MATH 2417) and (PHYS 1301 or PHYS 2325 or equivalent) and (BIOL 3361 or CHEM 3361). (4-0) Y

BIOL 4V00 Special Topics in Biology (1-6 semester credit hours) May be repeated as topics vary (9 semester credit hours maximum). Prerequisites: (BIOL 3301 and BIOL 3302) and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. ([1-6]-0) S

BIOL 4V01 Topics in Biological Sciences with Lab (1-6 semester credit hours) May be repeated as topics vary (6 semester credit hours maximum). Prerequisites: (BIOL 3301 and BIOL 3302) and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. ([1-5]-[1-5]) R

BIOL 4V40 Special Topics in Molecular and Cell Biology (1-6 semester credit hours) May be repeated as topics vary (9 semester credit hours maximum). Prerequisites: (BIOL 3301 and BIOL 3302) and (BIOL 3361 or CHEM 3361) or equivalent or instructor consent required. ([1-6]-[0-5]) S

BIOL 4V95 Advanced Topics in Biology (Individual Instruction) (1-6 semester credit hours) Individual study under a faculty member's direction. May be repeated for credit as topics vary. Instructor consent required. ([1-6]-0) S

BIOL 4V99 Senior Honors Research in Molecular and Cell Biology (3-6 semester credit hours) For students conducting independent research for honors theses or projects. Besides the university specifications, the student should contact the undergraduate advisor in biology for program requirements. May be repeated for credit as topics vary. Instructor consent required. ([3-6]-0) S


BIOLOGY, 3rd Ed./LIVE: Edmondson (Option 3)

Aug 25, 2021 at 9 am , runs for 32 weeks

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Teacher-Led & Graded by Becky Edmondson, B.S. Biology (Pre-Med):

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Measuring Biological Responses with Automated Microscopy

Christine C. Hudson , . Carson R. Loomis , in Methods in Enzymology , 2006

Meeting the Demand of High‐Throughput Screening with Transfluor

High‐throughput screening assays need to be predictive of the pharmacology of a test compound on its target, simple and easy to perform, robust, and automatable. The arrestinGFP translocation assay has been shown to meet all of these requirements. First, it has been used successfully to determine the pharmacology of known agonists and antagonists for several GPCRs ( Ghosh et al., 2005 Oakley et al., 2006 ). Next, the Transfluor assay is performed easily because it involves a few, basic steps ( Fig. 6 ). In contrast to other high‐content screening assays, which require primary and secondary antibody incubations and washes after cell fixation, no wash steps are necessary with Transfluor. In addition, all steps in the assay are automated easily by liquid‐handling robotics such as MultiDrops and MiniTraks, and detection of assay results has been validated on multiple, automated fluorescent imaging platforms ( Oakley et al., 2006 ). Furthermore, the assay is very reproducible and gives excellent screening statistics, including Z prime values above 0.5, ensuring an optimal screening assay window.

Fig. 6 . Transfluor screening protocol steps with associated liquid‐handling robotics.


Watch the video: Κατηγορίες μικροοργανισμών Λυκ (January 2023).