The cells in question have been fixed in 1% paraformaldehyde. The problem is that the hyphae intertwine upon germination, and it's difficult to observe individual conidia germination. I know that we could mount aliquots on slides, but I wouldn't be able to stop germination at precisely a certain time point.
Try adding a few drops of Tween 80; it will aid on avoiding clump formation.
Fungal species that is? so it would be important to know the size of the spore
Make the conidial suspension in 0.01-0.1 Tween 80 and then filter it trough miracloth.
First tap the fungal culture (germination spore) for few min and 2-4 drop of tween 80 and incubated at 4 degree/h
That's everyone for your suggestions. I'll give some of these a try. I have been collection conidia (8uM) in PBS 0.05% Tween20, but it seems like I can increase the concentration of tween 20 or try using tween 80. I guess since paraformaldehyde or 10% buffered formalin should permanently cross-link amine groups that adding tween would not undo anything. In other words, germination shouldn't start again. This fungal hyphae laughs at quick sonication, but I haven't tried sonication in a higher concentration of tween either.
.5-1percent Tween 50% +30% Formalin 40%+ 2% ethanol will easily will flour and separated from each other. Another make distil water suspension to germinate and collect them while germ tube comes out.Best of luck.
Ebrahim
Make a solution of 0.01-0.1% tween 80 or 20 in sterile distilled water, add some few ml to the inoculum plate and gently rock the plate and decant the spores for counting
As suggested above use 0.01-0.1% Tween 80 or 20 or any other nontoxic wetting agent such as sodium dioctyl sulfosuccinate or sodium laurel sulfate in sterile distilled water in Erlenmeyer flask containing some solid glass beads, add some inoculum, shake the flask vigorously to liberate the spores from fruiting bodies and to break the spore clumps than filter the fungal suspension through a layer of sterilized glass wool contained in a glass funnel.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA