The application discloses a kind of cotton
callus autophagy activity
visual detection technology, first using pretreatment buffer to incubate
callus, then using PBS buffer transition, using MDC
dyeing liquid is dyed, prepare
cell suspension, observation.The application increases the level of
cell autophagy by pretreatment induction, significantly improves the
fluorescence staining intensity, improves the contrast of specific dyed
autophagosome and background, so that
autophagy in
cell is more easily observed;Meanwhile, the pretreatment buffer has the effect of maintaining
osmotic pressure, keeping liquid environment pH stable and providing basic
nutrition, which can significantly improve the stability of
staining.The application uses PBS buffer to wash, which can avoid over-
staining and reduce the degree of background
clutter.The staining step of the application is more simple, with clear staining levels and
clear cell contrast, so that the autophagy phenomenon in
plant somatic embryogenesis process is efficiently visualized.