1.5 DAPI and CFW staining of dormant and non-dormant spores
The spore suspension to be tested was centrifuged 200 μL at 1500 rpm for
5 min, and the supernatant was discarded, and then centrifuged with PBS
(pH7.2) twice, and the supernatant was discarded. The washed spores were
fixed with 5% paraformaldehyde at 4 ℃ for 24 h, and then gently washed
with PBS for 3 min. The spores were suspended in 10 mL DAPI dye working
solution (1 μg/mL, Shanghai Jizhi Biochemical Technology Co., LTD) for
20 min. The stained samples were placed on slides, and the fluorescence
was analyzed under confocal laser scanning microscope with maximum
excitation wavelength of 340 nm and maximum emission wavelength of 488
nm. According to the instructions recommended by the reagent
manufacturer (Wuhan Purity Biotechnology Co., LTD.), the CFW dye drops
were added to the chlamydospore fixed with 5% paraformaldehyde, the
fluorescence intensity was adjusted, and the staining of the cell wall
was observed and photographed under a fluorescence microscope.