Simulation 1 - 3 (for Lab Practical One) Flashcards
What is the correct order of steps for a
Broth to Agar Plate.
Remove a loop full of bacterial culture from the broth tube, pass the mouth of the tube through the Bunsen burner flame, and replace the cap
Lift the lid of the agar plate slightly and streak the loop across the surface of the agar plate. Replace lid.
Sterilize inoculating loop, remove cap of culture tube, and pass mouth of culture tube through the Bunsen burner flame
Sterilize inoculating loop prior to replacing in loop holder
What is the correct order for
Broth to Broth Transfer
Remove a loop full of broth culture, pass mouth of broth culture tube through Bunsen burner flame, and replace cap on tube
Sterilize and cool inoculating loop, remove cap of culture tube, pass mouth of tube through Bunsen Burner flame
Pass mouth of broth tube through Bunsen burner flame and replace cap. Sterilize inoculating loop and return to loop holder
Remove cap of sterile broth tube, pass mouth of tube through the Bunsen burner flame, and insert inoculating loop into sterile broth
What is the proper use of Bunsen Burner
Be sure the gas valve is closed
Turn the rube in a clockwise direction until the air flow is almost complete closed
light a match and hold it off to one side of the top of the tube
Slowly turn on the gas by swiveling the gas valve until it is in line with the pipe. The gas should ignite
Adjust the tube and the needle valve (under base of the Bunsen burner) until a suitable flame has been achieved
Definition of Sterile
The freedom from the presence of viable microorganisms
Definition of Aseptic Technique
Combination of protocol that collectively maintain sterility, or asespsis, to prevent contamination of a patient or surface with microbes and infectious agents
What type of pathogen is B. Anthracis
Gram-positive, rod shaped bacterium
What is the correct laboratory BSL for B.Anthracis
BSL level 3
Why do you think Bacillus Anthracis is classified as Hazard grouped 3
It may be spread the community, but treatment is usually available
Which facility must we if we have an Ebola Sample?
BSL-4
Why is it important that a negative air pressure is maintained inside the Biosafety level 3 (BSL-3) lab
It minimized the chance of a microorganism leaving the lab
When should gloves and oversleeves be worn in a BSL-3 or CL3 lab
Gloves should be worn at all times, but over sleeves only when working with the cabinet
Once you have completed you training, which of these factors is NOT important with regard to your buddy Marie?
Marie should remain in the lab with you at all times
Which of these factors apply to Class 3 mode
Class 3 mode provides a physical barrier between the user and the microbes
What does HEPA stand for in a HEPA filter?
High - Efficiency Particulate Air
Should we put on the side panel in Class One mode? The side panel should be put on ______
So there is a stronger airflow into the front aperture of the cabinet
In class One mode, should the night panel be removed or attached?
Removed. You are protected by air drawn into the cabinet
As you can see, the status of the BSC is CLEAN. When should the CLEAN sign be used
When the cabinet contains nothing hazardous
When testing whether the powder sample contains the anthrax agent (B.anthracis) or not. What should you do first?
Isolate the DNA of the bacteria from the sample
Why is it important that sterility test is carried out
To ensure the culture is completely sterile before being removed from the lab
You have to be careful when working inside the biosafety cabinet. What should you do to clean the spill.
Perform fumigation
What should all work surfaces be cleaned with at the end of each working day?
5% biocleanse
The different personal protection equipments are put on and taken off in a specific area. Which of these statements is correct?
Overshoes are put on in the anteroom and taken off in the lab
Why do all staff need to sign in and out of the lab?
Only authorized people are allowed in the lab, a sign out sheet keeps track of this
When fumigating the lab, which of the following should not be carried out
Fridge and freezer doors should’ve be cleaned and opened
Calculation of Total magnification
Ocular Lens magnification X Objective Lens Magnification
Definition of Resolution
Reveals the outcome of the case and unpack the broader lessons to be learned
Factors Affecting Resolution
Wavelength and numerical aperture
What is the correct negative control to determine whether your reagents and tools are contaminated?
A. None; you only need to do a positive control when culturing microbes
B. Unculture Medium: if no colonies form on this plate, there is no contamination
C. Water: if colonies form on this plate, something was contaminated
D. Add Antibiotics to the sample: if no colonies form on this plate, everything was safe
B: Unculture Medium: if no colonies form on this plate, there is no contamination
After disinfecting work space, what’s your next action?
A. Take off your safety goggles
B. Go and have lunch
C. Turn Bunsen burner back on
D. Wash your hands
D. Wash your hands
What is 1-3
1: Eye piece (ocular lens)
2: Revolving nose piece (to hold multiple objective lenses)
3: Objective Lenses
What is 9, 4, 5
9: Mechanical stage
4: Coarse focus (large knob)
5: fine focus (small knob)
What is 6, 8, 7
6: stage (to hold the specimen)
8: Diaphragm
7: Illuminator
What the two no numbers (top to bottom)
Top: X-Y mechanical stage knobs (to move slide)
Bottom: Rheostat (to adjust light intensity)
Procedure of Fixing bacteria to a slide
Thin layer of specimen is spread on the slide (smear)
Slide is then briefly heated over the heat source
Procedure of Gram Stain Method
Step 1: Crystal Violet (primary stain added to specimen water)
Step2: Iodine (Mordant makes dye less solvable so it adheres to cell walls
Step 3: Alcohol: decolorizer washes away stain from gram-negative cell walls
Step 4: Safranin: counter stain allows dye adherance to gram-negative cell
Procedure of Acid Fast Satin Method
Primary, decolorizing, counterstaining
Acid-fast staining procedure would be categorized as a differential stain
Procedure of Endospore Stain Method
1: Place slide on beaker with boiling water
2: saturate slide with Malachite green
3: staring for 5-6 mins, add more water as it evaporates
4: Forceps to remove slide and cool slide
5; rinse slide with water to remove excess malachite green
6: Cover smear with safranin for 60-90 sec
7: rinse to remove excess safaris
8: blot slide with bibulous paper
What is the purpose of staining biological samples
Adding contrast and labeling specific structures
What does it mean to have parfocal objectives?
The sample remains in focus when the objectives are changed
Which of these factors is improved by adding immersion oil when using the 100x objective?
Resolution
Can Scientist use the sampled they have already stained with the Mallory stain to study goblet cell numbers and morphology?
Yes, the outline of the cell, the nucleus and mucus each have a different color
How to interpret Gram Stain results
Purple = Gram Positive
Pink/red = Gram Negative
Interpretation of Acid Fast Stain
Pink = Acid Fast Bacteria
Blue/purple = Non Acid Fast Bacteria
Interpret Endospore Stain
Endospore appear Bluish green
Non Endospore pink/red
Gram Stain
Acid Fast Stain
Endospore Stain
