Surface swipe
Suggested course of action:
- The surface may have been contaminated with endospores.
- Endospores germinate within 15–30 minutes, so work quickly to prevent germination.
- Each sample of the triplet set should be evaluated separately to avoid cross-contamination.
- Add water (0.5–1ml) to the tube.
- Vortex 15s
- Aliquot 10µl liquid sample to the center of each of the following I-F slide types: DAPI, SYTO, PI, Insti-Spore; or use an Insti-Fluor Combo2 test slide. (Click on the links to see step-by-step instructions.) Mark each test slide with the FPAT vial designation.
- Dry at 42C (10min), add 7µl immersion oil, apply coverslip and evaluate using epifluorescence microscopy.
- It’s always a good idea to test powder samples with Calcofluor M2R (Insti-Fluor CF) to test for the presence of Yeast.
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).