Introduction to Equipment

1. Introduction to Equipment Biotechnology Fall 2012

2. Lesson 1: Essential Questions • What are the most common pieces of biotechnology laboratory equipment? • What is aseptic technique? Why is it important? What happens when we do not practice it?

3. Lesson 1: Learning Objectives • By the end of today, you should be familiar with the most common pieces of lab equipment • You should know what each piece of equipment is and what it is used for • In addition, you should know what aseptic technique is and why it is important in the biotechnology laboratory

4. Measurements • In the science laboratory we use the metric system • To measure volume= liter • To measure mass or weight= gram • To measure length= meter • However, Biotechnology often works on the milli and micro-scales (103 and 106)

5. Volume • To measure large volumes (greater than 25 ml) you would use a graduated cylinder • They come in several sizes • They are very accurate • Measure to meniscus

6. Volume • Larger volumes (greater than 25 ml) should be made in beakers and flasks • These should not be used to measure, as they are not very accurate

7. Volumes • Small amounts are usually measured with pipets • There are two types • Glass pipets (2-25 ml) • Micropipets (0.2 – 1 ml)

8. Mass or Weight • We will always use the measurement of grams • The scales we have are very accurate and expensive

9. Centrifuge • Provide a centrifugal force to move liquid to the bottom of a tube • Regular • Micro

10. Aseptic Technique • Microbes are ubiquitous • Reduces the chances of contaminating yourself, others, or the experiment • Involves the following: • Sterilizing all equipment and supplies • Keeping all equipment and supplies sterile • Never touch anything with your bare hands • Everything that touches an experimental substance should be sterile • Keeping containers closed as much as possible • Disinfecting your hands and bench frequently

11. Aseptic Technique • What happens when you are not sterile? Lets see with the use of a “deadly virus” • Everyone grab 1 tube of powder • Glove up • Squirt some glycerol on your glove and really rub it in • Pour your tube of powder all over your hand • Shake hands with 3 people • Spread out and shake with 3 people not around you • View your hand under UV light • What happened?

12. Lesson 2: Essential Question • How do we convert grams, meters, and liters correctly?

13. Lesson 2: Learning Objectives • Know how to convert grams, meters, and liters correctly

14. Conversions

15. Conversions • When you go to a smaller measurement, you move the decimal to the right • When you go to a larger measurement, you move the decimal to the left • Lets try these!

16. Lesson 3: Essential Question • How do we properly use a glass pipet to transfer liquids?

17. Glass Pipets (1-25 ml) • 4 Different Sizes • Attach the Pipet and adjust volume by moving the wheel • Watch closely while pipeting • Must be disposed of in the sharps container

18. Lab • Practice using graduated cylinders, scales, and glass pipets • Follow the protocol given to you • Make sure to be as accurate as possible and use aseptic technique at all times • Dispose of everything in its proper place • Don’t forget to clean up when you are finished!

19. Lesson 4: Essential Question • How do we use a micropipet correctly?

20. Lesson 4: Micropipets

21. How to Use a Micropipet • To draw up a solution • Rotate the volume adjuster to the desired volume • Place the appropriate tip on the barrel • Press the plunger to the first stop • Insert the tip into the solution • Draw up the solution by gradually releasing the plunger • Check to see that there are no air bubbles present in the tip

22. How to Use a Micropipet • To expel the solution • Touch the tip against the side of the tube or container into which the solution will be expelled • Press the plunger to the second stop to make sure everything is expelled • Remove the pipette tip from the container with the plunger still pressed • Release the plunger slowly • Eject tip into the appropriate waste container by pressing the tip-ejection button

23. Care of the Micropipet • The following rules should be observed at all times: • Never place a pipette near the edge of the bench where it can get knocked over • Never rotate the adjuster below or beyond the designated ranges • Never immerse the pipette barrel in liquid • Never lay the pipette down or hold it horizontally when there is liquid in the tip • Always wipe the pipette barrel when you are done • Always use the appropriate tip for each pipette

24. Lab • Practice using the micropipet • Follow all instructions carefully • Be sure to use proper aseptic technique at all times

25. Lesson 5: Essential Question • How do we make solutions in the biotechnology laboratory?

26. Math Skills Needed for Biotech • You must know how to calculate solutions using the following equations! • You will do this every day in class • Like conversions, they will not go away, YOU MUST LEARN THEM NOW!!!!! • C1V1=C2V2 • Mass/Volume

27. Solute/Solvent= Concentration • Solutes (usually measured in g) • Solvents (usually measured in ml) • Mixed together until solute dissolves in solvent solute solution concentration =

28. C1V1=C2V2 • (starting concentration)(starting volume)=(final concentration)(final volume) • Solve for any of the variables, as long as you know the other 3 • How do we actually make the solution once we have the answer?

29. Mass/Volume Equations • The mass of solute in the amount of solvent • Calculates the amount of solute needed per amount of solvent Mass/volume ___________ _____________ __________________ concentration desired X volume desired = mass of solute needed % Mass/volume __________ ____________ _________________ conc desired (decimal) X volume desired = mass of solute needed

30. Examples: Mass/Volume To make 100 mL of a 5 mg/mL hemoglobin solution ___________ _____________ _________________ concentration desired X volume desired = mass of solute to be weighed 5 mg/mL100 mL500 mg = 0.5 g concentration desired X volume desired = mass of solute to be weighed

31. Examples: % Mass/Volume To make 100 mL of a 3% CuSO4 solution 3 % = 0.03 __________ ____________ _________________ conc desired (decimal) X volume desired = mass of solute needed 0.03 g/mL100 mL3 g CuSO4 conc desired X volume desired = mass of solute needed