Introduction to Equipment

Introduction to EquipmentBiotechnology Fall 2012Lesson 1: Essential QuestionsWhat 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? Lesson 1: Learning ObjectivesBy the end of today, you should be familiar with the most common pieces of lab equipmentYou 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 laboratoryMeasurementsIn the science laboratory we use the metric system

To measure volume= literTo measure mass or weight= gramTo measure length= meter

However, Biotechnology often works on the milli and micro-scales (103 and 106)VolumeTo measure large volumes (greater than 25 ml) you would use a graduated cylinderThey come in several sizesThey are very accurateMeasure to meniscus

VolumeLarger volumes (greater than 25 ml) should be made in beakers and flasksThese should not be used to measure, as they are not very accurate

VolumesSmall amounts are usually measured with pipetsThere are two typesGlass pipets (2-25 ml)Micropipets (0.2 1 ml)

Mass or WeightWe will always use the measurement of gramsThe scales we have are very accurate and expensive

CentrifugeProvide a centrifugal force to move liquid to the bottom of a tubeRegular Micro

Aseptic TechniqueMicrobes are ubiquitousReduces the chances of contaminating yourself, others, or the experimentInvolves the following:Sterilizing all equipment and suppliesKeeping all equipment and supplies sterile Never touch anything with your bare handsEverything that touches an experimental substance should be sterileKeeping containers closed as much as possibleDisinfecting your hands and bench frequently

Aseptic TechniqueWhat happens when you are not sterile? Lets see with the use of a deadly virusEveryone grab 1 tube of powderGlove upSquirt some glycerol on your glove and really rub it inPour your tube of powder all over your handShake hands with 3 peopleSpread out and shake with 3 people not around youView your hand under UV light

What happened? Lesson 2: Essential QuestionHow do we convert grams, meters, and liters correctly?Lesson 2: Learning ObjectivesKnow how to convert grams, meters, and liters correctlyConversions

ConversionsWhen you go to a smaller measurement, you move the decimal to the rightWhen you go to a larger measurement, you move the decimal to the left

Lets try these! Lesson 3: Essential QuestionHow do we properly use a glass pipet to transfer liquids?Glass Pipets (1-25 ml)

4 Different SizesAttach the Pipet and adjust volume by moving the wheelWatch closely while pipeting Must be disposed of in the sharps containerLabPractice using graduated cylinders, scales, and glass pipetsFollow the protocol given to youMake sure to be as accurate as possible and use aseptic technique at all timesDispose of everything in its proper place Dont forget to clean up when you are finished! Lesson 4: Essential QuestionHow do we use a micropipet correctly? Lesson 4: Micropipets

How to Use a MicropipetTo draw up a solutionRotate the volume adjuster to the desired volumePlace the appropriate tip on the barrelPress the plunger to the first stopInsert the tip into the solutionDraw up the solution by gradually releasing the plungerCheck to see that there are no air bubbles present in the tip

How to Use a MicropipetTo expel the solutionTouch the tip against the side of the tube or container into which the solution will be expelledPress the plunger to the second stop to make sure everything is expelledRemove the pipette tip from the container with the plunger still pressedRelease the plunger slowlyEject tip into the appropriate waste container by pressing the tip-ejection button

Care of the MicropipetThe following rules should be observed at all times:Never place a pipette near the edge of the bench where it can get knocked overNever rotate the adjuster below or beyond the designated rangesNever immerse the pipette barrel in liquidNever lay the pipette down or hold it horizontally when there is liquid in the tipAlways wipe the pipette barrel when you are doneAlways use the appropriate tip for each pipette

LabPractice using the micropipetFollow all instructions carefullyBe sure to use proper aseptic technique at all times

Lesson 5: Essential QuestionHow do we make solutions in the biotechnology laboratory? Math Skills Needed for BiotechYou must know how to calculate solutions using the following equations!You will do this every day in classLike conversions, they will not go away, YOU MUST LEARN THEM NOW!!!!!

C1V1=C2V2Mass/VolumeSolute/Solvent= ConcentrationSolutes (usually measured in g)Solvents (usually measured in ml)Mixed together until solute dissolves in solvent

concentration =solute solutionC1V1=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? Mass/Volume EquationsThe mass of solute in the amount of solventCalculates the amount of solute needed per amount of solvent

___________ _____________ __________________concentration desired X volume desired = mass of solute needed __________ ____________ _________________conc desired (decimal) X volume desired = mass of solute neededMass/volume% Mass/volumeExamples: Mass/VolumeTo make 100 mL of a 5 mg/mL hemoglobin solution ___________ _____________ _________________concentration desired X volume desired = mass of solute to be weighed 5 mg/mL 100 mL 500 mg = 0.5 gconcentration desired X volume desired = mass of solute to be weighedExamples: % Mass/Volume __________ ____________ _________________conc desired (decimal) X volume desired = mass of solute neededTo make 100 mL of a 3% CuSO4 solution3 % = 0.03 0.03 g/mL 100 mL 3 g CuSO4conc desired X volume desired = mass of solute needed