This invention discloses a rapid method for detecting
calcium signals in undifferentiated and differentiated spermatogonia. The method uses a mild
enzymatic digestion method to prepare a testicular single-
cell suspension. Indo-1 AM, Thy-APC, and c-kit-PE are used for
staining and labeling. The
cell population to be analyzed is determined based on the difference between the lateral and forward scattering areas of the cells in the testicular
cell sample. Then, Thy-APC and c-kit-PE are used to
label undifferentiated and differentiated spermatogonia, respectively, to further select specific cell groups. Finally, the
fluorescence intensity changes of indo-1 AM in each specific cell subpopulation are detected, thereby obtaining the changes in
calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. This method can rapidly analyze the
calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. The method is simple and convenient, and provides a foundation for identifying spermatogonial dysfunction caused by calcium
signal imbalance.