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.