Currently developing a rapid FISH method for the specific detection of Bacillus cereus, however our traditional methods of fixation methods fail to work (despite following previous research paper methods). Looking for any recommendations outside of formaldehyde/formalin, or glutaraldehyde (introduces auto-fluorescence). I have been considering SDS, since this is a protein disruptor and may have a positive effect on disrupting the peptidoglycan cell wall - any recommendations of concentrations would be appreciated.
Thanks in advance!
Hello Edward David Ash ,
I have used in the past methanol or acetone 100%. Indeed they act as both fixative and permeabilization agents making them a good alternative to PFA. The time of incubation is tipically 5-10min with these reagents.
However, depending on the structures you want to visualize, there could be better alternatives.
I hope this helps.
Best,
Mario
I did use cold (-20C) methanol or acetone for this kind of fixation for 10 minutes in one of my experiments
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