The invention belongs to the technical field of
sperm acrosome detection, and discloses an improved
crystal violet
staining method for detecting
sperm acrosome. The method comprises the following steps: preparing an improved
gelatin film; liquefying collected fresh seminal fluid, subjecting seminal fluid to
centrifugation, discarding seminal
plasma, suspending precipitates in a
phosphate buffer solution, placing the
buffer solution in a thermotank, evenly paving
sperm suspension on the
gelatin film, incubating the sperms in a thermotank with a temperature of 37 DEG C, after 30 minutes, takingout sperm suspension for microscopic examination; dropwise adding a
crystal violet
dyeing solution on a fixed smear, carrying out
dyeing for 1 to 3 minutes, washing the smear by water to remove excessdyes; dropwise adding a Lugol
iodine solution, after one minute, washing the smear by water, then dropwise adding
alcohol (95%) to perform color separation, shaking the
glass slide until the purple color is not faded, washing the smear by flowing water, and carrying out microscopic examination after
drying. A simple and rapid
dyeing medium adopted by the
crystal violet
staining method can enhancethe
cell dyeing affinity, through a discoloring treatment, the binding strength between a fuel and a polluted object can be measured, the
structural difference of cells can be discriminated, and thesituation of sperm
acrosome is well displayed.