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Med Microbiology
Stains - Coggle Diagram
Med Microbiology
Stains
Acid-fast Bacilli stain
Procedure
- Add a drop of saline on slide (frosted facing up)
- Take colony from 4th quadrant
- Mix colony with drop of saline and heat fix with Bunsen burner.
- Flood slide with carbon fuchsin and heat it for 5 minutes. Don't boil the stain. Add stain if necessary.
- Wash the slide with tap water.
- Flood the slide with 3% acid alcohol for 2 minutes.
- Wash the slide with tap water.
- Add methylene blue to slide for 1 minute.
- Wash the slide and allow to be air dried.
- Observe under microscope.
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Gram Stain
Procedure
- Add a drop of saline on slide (frosted side facing up)
- Take a single colony with sterile inoculating loop.
- Mix colony with saline and heat fix with Bunsen burner.
- Cover the slide with crystal violet for 1 minute.
- Dilute the slide with a gentle stream of water.
- Cover the microscope slide with iodine solution for 1 minute.
- Wash the slide with a gentle stream of water.
- Cover microscope slide with acetone for 30 seconds.
- Quickly rinse the slide with water.
- Cover the slide with safranin for 1 minute.
- Rinse the slide with water and observe under microscope
What is observed?
Gram-positive bacteria:
Purple or blue colour
Gram-negative bacteria:
Pink or red colour
Gram-Positive bacteria examples:
- Staphylococcus sp
- Streptococcus sp
- Clostridium sp
Gram-Negative bacteria examples:
- Escherichia coli
- Pseudomonas
- Salmonella
- Shigella
- Klebsiella
- Enterobacter
- Yersinia
Capsule stain
Procedure
- Add 1 drop of Indian ink to the slide
- Take a single colony with a sterile inoculating loop and mix well with the stain
- Leave slide to air dry
- Flood slide with crystal violet for 1 minute.
- Slightly rinse the slide with water and air dry slide.
- Observe the slide
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Endospore stain
Procedure
- Add a drop of saline on slide (frosted side facing up)
- Take a single colony with sterile inoculating loop.
- Mix colony with saline and heat fix with Bunsen burner.
- Cut a square blotting paper and cover on the smear.
- Slide put to steam while adding malachite green stain onto the blotting paper to keep the paper moist. Steam for 5 minutes.
- Wash slide with tap water.
- Add 0.5% safranin to the slide and leave for 30 seconds.
- Wash slide with tap water.
- Observe slide under microscope
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Albert Stain
Procedure
- Add a drop of saline onto a slide
- Take single colony with sterile inoculating loop
- Mix the colony with the saline drop well and heat fix the slide.
- Flood slide with Albert stain A for 7 minutes.
- Drain the excess stain and directly flood the slide with Albert stain B for 2 minutes. Do not rinse off Albert stain A.
- Wash the stain with tap water.
- Air dry the slide and observe under microscope.
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