Laser-scanning microscopy as applied to mouse early embryos: Cytometry and analysis of cell morphology

Abstract

This paper updates our knowledge on quantitative laser scanning microscopy and summarizes the capabilities of this method as applied to cytometry and analysis of cell structure of the mouse early embryo and the oocyte. This method requires a stack of optical sections obtained as Z-series with subsequent 3D reconstruction. This approach was used for visualization of the 3D cell model, measurement of the cell volume and surface area, as well as a study of the cell interior via optical sections. To maintain the dimensional characteristics of embryos or oocytes the method of sample preparation involved the following consequent steps: rapid cryofixation, low-temperature dehydration, infiltration by optically transparent mounting media, and laser-scanning microscopy. This strategy enables volume measurement, even in the case of a single cell within the multicellular system of the mouse early embryo.