Quantitative Mast Cell Analysis

The stained tissue sections were observed on a ZEISS Axio Imager.Z2 that was equipped with a Zeiss alpha Plan-Apochromat objective 100×/1.46 Oil DIC M27, a Zeiss Objective Plan-Apochromat 150×/1.35 Glyc DIC Corr M27, and a ZEISS Axiocam 712 color digital microscope camera. The captured images were processed with the software programs, “Zen 3.0 Light Microscopy Software Package”, “ZEN Module Bundle Intellesis & Analysis for Light Microscopy”, “ZEN Module Z Stack Hardware” (Carl Zeiss Vision, Aalen, Germany), and submitted with the final revision of the manuscript at 300 DPI. Photomicrographs were obtained in some cases with a confocal microscope, Nikon D-Eclipse C1 Si based on Nikon “Eclipse 90i”. The number of mast cells in the different areas of the ovary was determined per mm² of tissue, using open-source software for the digital pathology image analysis, QuPath [90 (link)] (Bankhead P., et al., 2017) (Table 1).