Three-Dimensional Confocal Analysis of Chromosome Positioning Coupled with Immunofluorescence in Mouse Sperm Nuclei.

We describe here a in situ fluorescence hybridization (FISH) strategy coupled with a fluorescent immunocytochemistry approach followed by confocal analysis and reconstruction (2D/3D) as a powerful tool to analyze the location of chromosomes in the sperm nucleus using the mouse sperm as a model. Already, the two-dimensional (2D) analysis of FISH and immunofluorescence data reveal the location of chromosomes as well as the different markings on the spermatic nucleus. In addition, a good 3D rendering after Imaris software processing was obtained when Z-stacks of images were acquired over a defined volume (10 μm × 13 μm × 15 μm) with a sequential scanning mode to minimize bleed-through effects and avoid overlapping wavelengths. PMID: 32822037 [PubMed - as supplied by publisher]
Source: Reproductive Biology - Category: Reproduction Medicine Authors: Tags: Methods Mol Biol Source Type: research