To analyze the teaching of mouse kidney transparent fluorescent specimen preparation and light-sheet fluorescence microscopy imaging skills.

The participants included 2 doctoral students and 2 other scientific researchers, and the training instructor was a senior renal biological imaging technician. The training lasted for 31 days. At different time points during the training, the success rate of fluorescent antibody injection in mice, preoperative mortality rate, cardiac perfusion bleeding rate, success rate of kidney sampling, kidney transparency degree, and kidney fluorescence imaging effect were evaluated and statistically analyzed.

Compared with those of the first day of training, the success rates of fluorescent antibody injection and kidney harvesting were higher on the fifth day of training, while the preoperative mortality rate of mice and the bleeding rate during cardiac perfusion were lower (all P<0.01). On the 10th, 20th, and 30th days of training, the transparency of the kidneys gradually improved. And on the 30th day, the transparency of the mouse kidneys met the standard. On the 31st day of training, the imaging effect of the light-sheet fluorescence microscopy for the participating students also met the standard.

A 31-day analytical teaching program covering 8 experimental operation skills and 5 teaching essentials enabled the trainees to master the preparation of transparent fluorescent specimens of mouse kidneys and the imaging skills of light-sheet fluorescence microscopy.