Today s Goals

1. Today�s Goals Determine the most appropriate tool for measuring specific volumes Describe how to select, set, and use a variety of micropipets within their designated ranges to accurately measure small volumes Demonstrate proper use of pipets and micropipets to properly dispense solutions Convert between units of measure using appropriate conversion factors

2. Measuring Volumes in a Biotechnology Facility Volume is a measurement of the amount of space something occupies Volume is measured in Liters (L) Milliliters (mL) Microliters (mL) How many microliters are in a milliliter? How many microliters are in a liter? Different tools are used to measure volume Graduated cylinder Pipet (seriological) Micropipet

3. Volume Units Often volumes are measured in one unit of measurement and reported in another Converting between metric units Conversion factor We will discuss later in this lesson

5. Using Seriological Pipets Measuring units smaller than 10 mL requires a pipet.

6. What are they? Used for pipetting liquids ranged from .1 mL to 10 mL to make solutions. Long glass tube able to hold a set amount of liquid Come in sizes, 1mL, 2 mL, 5mL, and 10mL. Pumps are used with tubes to take up and dispense liquid.

7. How to read them Usually in intervals of 10 For example: .1,.2,.3� 1mL Each pipette has a certain pump to use 1mL and 2mL use blue pump 5 mL and 10 mL use the green pump Larger pipettes are graduated differently Smaller pipettes are graduated from the bottom to top. Larger pipettes are also graduated top to bottom Pipette tip is included in interval measurements Demonstration

8. How to use them Firmly put glass tube in pump and slightly twist to seal. Do not apply to much force Roll wheel up with thumb to take up liquid Roll wheel down to dispense liquid Trigger is used to completely dispense Take the glass tube out of pump carefully Wipe excess liquid off tip using top of beaker If TD is on glass, do not force extra liquid out Demonstration

9. Demonstration of Use of Serological Pipets Why is it important? Video- Biotech DVD 1 & 2 Points to remember for practical Choose the smallest pipette for the volume you need to transfer Pick up the tube and pipettes at eye-level Read the volume of liquid from the bottom of the meniscus for best accuracy Pick up the tube into which you will pipet the liquid and slowly lower the plunger

10. Practice Questions What equipment would you use to measure the following, most accurately? 125 ml 10 ml 4 ml 6 ml 60 ml

11. Micropipetting

12. Uses in the Lab Loading wells in electrophoresis and page gels PCR reactions Broth culture Bacteria on Agar Plates

14. Types 2-20ul 20-200ul 100-1000ul

17. Observe the 3 sizes of micropipets and tips we have in class

19. The �How to� Use the correct micropipette depending on the amount you need. P-10 measures 0.5-10�L P-20 measures 2-20�L P-100 measures 10-100�L P-200 measures 20-200�L P-1000 measures 100-1000�L Choose the pipette that is most accurate For example, if you want to measure 20uL, use a P-100 because the P-20 maxes out to 20uL and is less accurate.For example, if you want to measure 20uL, use a P-100 because the P-20 maxes out to 20uL and is less accurate.

20. Set the micropipette dial to the desired volume. Obtain a box of pipette tips of the appropriate size small white tips for the P-10, p20 medium yellow tips for the P-100, and P-200 large blue tips for the P-1000

21. Taking Up a Sample Before picking up the micropipette, open the cap or lid of the tube from which you are taking fluid. Push the end of the pipette into the proper-size tip Close the tip box to prevent contamination.

22. Hold the micropipette in one hand at a 45 degree angle This way, contaminants from your hands or the micropipette won�t fall into the tube Hold the container you�re drawing from in your other hand. Hold both up at about eye level.

23. Push down on the plunger and STOP when you feel the first �stop.� Hold the plunger down in that position while you place the tip of pipette into the solution you are pipetting.

24. Slowly release the plunger while the tip is still under the solution�s surface Congratulations! You�ve drawn fluid into the tip! Now onto expelling it�

25. Expelling a Sample Keep holding the pipette with one hand at a 45 degree angle With your other hand, open the cap of the tube container you�re filing.

26. Hold the container and your micropipette in separate hands and at about eye level. Gently touch the inside wall of the container with the micropipette tip to get rid of excess liquid. Also, this creates a tiny surface-tension effect that helps draw the fluid out of the tip!

27. Slowly push the plunger down to the first stop and then continue to the second stop so that all the fluid is out of the tip. This is to ensure that the entire sample has been released Hold the plunger all the way down at the second stop until you have removed the pipet from the collection tube This way, you won�t draw any liquid back into the tip

28. Change the tip for each new solution (in this case color) being used. To do this, press the ejector button on top of the micropipet and the tip will shoot off. Throw out used tips in waste beakers or the garbage can!

29. WHAT IS DIALING DOWN? tips are disposable and usually intended for one use.

30. Demonstration: Proper use of Micropipets Why is it important? DVD- Reading, setting, proper use #3,4,5,6

31. Practice Questions What equipment would you use to measure the following, most accurately 25 mL 10 mL 1 mL 1000 mL 1 ml 2 ml 175 mL

32. What are Metric Conversions? Sometimes we need to change from one measurement to another This is usually done because the unit that is used is either too big or too small for that type of measurement.

33. TOO BIG!!! 525,568 millimeters Sometimes the number is too big for the unit being measured.

34. TOO SMALL 0.00654 km Sometimes the number is too small for the unit being measured

35. How Do We Fix This Problem? We can correct these problems by doing a process called Metric Conversion. When you do a Metric Conversion, you are changing from one unit of measurement to another.

36. Conversion of Metric Measurements-Review Basics To convert from a larger unit to a smaller unit- Multiply Decimal point moves to the right Ex. 1L = 1000 ml To convert from a smaller unit to a larger unit- Divide Decimal point moves to the left Ex. 100 mls - 0.1 L

37. Length Units Most important to know for Biotech Lab Kilometer (km) Meter (m) Centimeter (cm) Millimeter (mm) Micrometer (mm) How many micrometers in a millimeter? 1000 How many millimeters in a centimeter? 10

38. Mass Units Most important to know for Biotech lab Kilogram (kg) Gram (g) Milligram (mg) Microgram (mg) How many milligrams in a gram? How many micrograms in a gram?

39. Volume Units Most important to know for Biotech Lab: Liter (L) Milliliter (ml) Microliter (mL) How many milliliters in a liter? How many microliters in a milliliter? How many microliters in a liter?

40. Practice Questionswipe board Convert the following

42. Now it is time for you to practice�.. 1 worksheet (front & back): Complete in class or for homework. Next Class: Reread Lab 3 A & B Review metric worksheet