Microscope
Procedure of Labs
Purpose/Important concepts of Labs
Enzymes/Reactions/pH Indicators
Biochemical Tests
100
Function of the Objective lens
primary lense that magnifies specimen
100
We heat fix because...
Heat fixing denatures bacterial enzymes' causing autolysis, enzymes digesting cell parts that cause the cell to break as well as adhering it to slide plus to make sure bacteria are dead
100
The lab we use to differentiate gram positive from gram negative bacteria....
gram stain
100
Carbohydrate fermentation uses the pH indicator...
phenol red
100
This is a test of....
Gelatin Test that's testing for the hydrolysis of gelatin
200
Holds the slide in position
stage
200
Preparing a smear we....
1.Wash slide with soap
2. Label slide (circle in center of slide/place your name+ bacterial species) Flip slide over
3. Place drop of DW in circle if bacterial species is dry
4. Aseptic technique (sterilize loop 5 seconds till red hot, cool loop 30 seconds, aseptically open bacterial culture (open culture Using clam shell + gather bacteria on loop and transfer to slide) sterilize loop by heating loop in incinerator for 5 second and place on side of incinerator to cool)
5. Let slide dry of slide warmer
6. Heat fix slide
200
Flocculant growth is...
describes unevenly dispersed growth containing suspended aggregates and clumps
200
The reaction and enzyme for the starch test is...
starch--amylase-->glucose + glucose
200
The MR and VP in MRVP test for....
MR tests for presence of acid (methyl red)
VP tests for presence of acetoin (VP-A & VP-B)
300
Carrying microscope, we....
have one hand on arm and one on base with microscope close to chest securely
300
To do a gram stain....
Aseptically prepare smear, air dry slide on slide warmer &heat fix
1. Flood smear with Crystal violet
2. Rinse with DW above smear until clear
3. Flood swear with Gram's iodine
4. Rinse with DW
5. Decolorize with Gram's alcohol holding slide at angle. Use dropper and place decolorizer to run through smear till clear
6. Rinse with DW
7. Counterstain with safranin
8.Rinse with DW
9. Air dry (slide warmer)
300
A large zone of inhibition means....
Displays the microbe is susceptible to antibiotic making the antibiotic effective against microbes.
300
The test Phenylalanine deamination uses the enzyme...
Deaminase
300
A positive and negative urea test is...
(+) = fuchsia/ hot pink
(-) = not fuchsia
400
True or False
We drag microscope across table to help view the specimen better
False
400
For a capsule stain we...
1. Begin w/ drop Congo red stain at end of clean slide
2. Sterilize loop& aseptically remove small amount of bacteria culture mixing it with the Congo red and resterilize loop
3. Use 2nd clean slide to draw drop across to other end
4. Air-dry (do NOT heat-fix)
5. Flood w/ Maneval's stain
6. Gently rise with DW
7. Allow to air dry
400
Obligate aerobe grow on aerobic plate, anaerobic plate, and thioglycolate like....
-On Aerobic Petri plate: Little growth
-On Anaerobic Petri plate: Little/No growth
-On thioglycolate broth: Growth on the top
400
The equation for catalyze is....
2H2O2 --> 2H2O + O2
400
In the nitrate test if A&B reagents are added to test tube but it is colorless, so we add zinc, and it remains colorless this means...
nitrate was reduced to nitrogen gas
500
We never use_______on 40X objective
coarse focus as it will drive objective into slide
500
To do a tube to tube transfer....
1. Always label media
2.Sterilize loop
3. Open the tube using pinky finger of the hand holding loop
4. Flame the lips of the tube using a Busen Burner
5. Inoculate broth (see film of broth)
6. Flame the lips of the tube using a Busen Burner
7. Replace cap of tube
8. Grab second tube
9.Open the tube using pinky finger of the hand holding loop
10.Flame the lips of the tube using a Busen Burner
11. Place Inoculated broth and swirl around
12. Flame the lips of the tube using a Busen Burner
13. Replace cap of tube
14. Sterilize Loop
500
Mannitol salt is differential and selective because....
*Selective: encourages growth of gram positive while discouraging gram negative using high osmotic environment
*Differential: has a pH indicator to differentiate which species ferments the carbohydrate mannitol producing organic acid
500
Decarboxylation removes ____ from____ using_____
Decarboxylation removes a carbon dioxide from an amino acid using decarboxylase
500
A positive citrate plate is created by...
CO2 accumulated and reacted with Na+ and H20 to produce an alkaline compound making the medium pH become basic causing the pH indictor to change to Prussian blue
