The invention discloses a method for detecting openness of permeability transition pores (
MPTP) of peony
pollen mitochondria under physiological conditions by
computer assistance, which comprises the following steps: preparing
Calcein-AM
staining working solution and CoCl2 solution, preparing
pollen cells to be stained, grouping
staining,
staining the
pollen cells, preparing stained pollen flakes, observing, shooting, and analyzing by computer-aided
software (Aivia). And pollen
MPTP open
fluorescence intensity analysis is carried out. The method has the beneficial effects that the
dyeing concentration and steps of
Calcein-AM / CoCl2 are optimized aiming at peony pollen cells, the pollen cells and the
fluorescence intensity of the pollen cells on each image in different
dyeing groups are accurately identified by combining computer-aided analysis, pollen self-
luminescence interference is eliminated by taking an undyed group as a reference, and the detection accuracy of the pollen cells is improved. By creating an object classification model based on computer
deep learning,
fluorescence intensity intervals of
MPTP dyeing are accurately divided, and a
workflow is created to realize
batch processing. According to the method disclosed by the invention, the opening accuracy and
visibility of the
Calcein-AM / CoCl2 staining method for determining the MPTP are greatly improved, and batch
image analysis and
processing are realized.