Hi everyone,
I'm looking for a relatively simple viability test method to distinguish between live and dead spores (fungi i.e. Botrytis or Penicillium). I need to be able to do this using a compound/optical/light microscope and a counting chamber. I was looking at trypan blue but the warning labels (carcinogen) diverted my attention to methylene blue. Can anyone give me some background on these or other test methods? For now, I want to try and avoid expensive equipment (fluorescent microscopes and flow cytometry). Any suggestions to alternative systems that might be cheaper?
Thank you,
Pieter
Dead spore is rare spore is very strong and can survive in different conditions if not heat treated. Best method to test is use nutrient media to see the growth.
Hi Pieter,
sadly everything that binds to nucleic acids is at least mutagenous.
For live-death staining I used a staining kit by Molecular Probes (Invitrogen) and a fluorescence microscope.
Usually these stains come in high concentrations so they last quite long. Then the price is tolerable.
Dead spore is rare spore is very strong and can survive in different conditions if not heat treated. Best method to test is use nutrient media to see the growth.
Image analysis method for assessing the viability and the germination characteristics of fungal spores in submerged cultures. Look for swelling, germ tube formation, and germ tube elongation. Alternatively, preparation from 25 μg of living spores phosphatase or glucosidase to give a positive fluorescence test for hydrolysis of methylumbelliferyl-phosphate or methylumbelliferyl-glucoside at 37 C in 2 h.
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