Liquid in a tube
The sample is Liquid
- If the sample is liquid, then there is a small chance for Bacillus endospores if the liquid has a high osmolality. Use an Insti-Fluor Combo2 slide to test for bacteria and possible endospores.
- If the sample is high solute liquid (salt, sugar, etc) then endospores can stay ungerminated, and bacterial vegetative cells will not grow. Use an Insti-Fluor Combo2 slide to test for endospores as well as vegetative cells.
- Procedure: Agitate the sample, let settle 3s, add 10µl to the center well of a test slide
- Dry at 42C
- Add 7µl immersion oil
- Add coverslip
Observe
Scenario: Three famous ballparks have recently sold soft drinks that made people sick. Immediately afterward the Boston Globe, Chicago Times, and the St. Louis Post-Dispatch received an email indicating that the Sprite from one vendor at each stadium was contaminated with E.coli bacteria. Your CST has been called in to assist local authorities in the analysis of each of these samples to determine whether the soft drink contained bacteria.
For this test use either liquid probes or Insti-Fluor Combo2 test slides.
- Sample ALS-FPAT-D: a vial containing Sprite samples from Fenway Park.
- Sample ALS-FPAT-E: a vial containing Sprite samples from Wrigley Field.
- Sample ALS-FPAT-F: a vial containing Sprite samples from Dodger Stadium.
ANALYSIS OBJECTIVES: The Incident Commander requests that you use fluorescence microscopy to determine which sample(s) (if any) contain bacteria.
Special instructions:
- You will receive a scenario indicating a suspected bio-agent (E. coli for example). They also give you three different sample vials. You should look for ANY bio-agent in any vial and not limit yourself to the suspect biological. Keep to the “Closed Set”, but always suspect that there will be something different in each different vial.
- It is unlikely that there is more than one bio-agent per sample vial. Thus, if you find a lot of yeast (for example) in one vial, don’t spend a lot of time looking for another bio. It’s usually ONE Bio per sample vial.
- You will almost always find “bacteria” and “starch” in any sample, as it is a common contamination. Look for an overwhelming number of bio-agents in the samples and report that.
- The only definitive fluorescence test for fungi is Calcofluor M2R (Insti-Spore CF).