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. 2016 Feb 19;19(1):117–124. doi: 10.4103/1008-682X.168684

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Copyright: © 2017 AJA, SIMM & SJTU

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

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Representative images of the staining results for spermatozoa. The bar represents 5 µm. (a) aniline blue staining of three populations of spermatozoa: b: deprotaminated, c: semi-deprotaminated and d: normal, with a proper protamine: histone ratio; light microscopy, ×1000 magnification; (b) TUNEL assay involving the two populations of spermatozoa: light-green (with fragmented DNA) and blue (only DAPI as a counterstain, without DNA fragmentation); fluorescent microscopy, ×1000 magnification; (c) examples of the two-dimensional thin-layer cellulose chromatography (TLC) analysis of [5’³²P]-labeled deoxynucleotides obtained after the enzymatic hydrolysis of DNA. dNp – deoxynucleoside monophosphates: A: adenine; C: cytosine; G: guanine; T: thymine and m⁵C: 5-methylcytosine. Three different values for the R coefficient are presented; (d) example of the immunofluorescent staining (IF) of spermatozoa. Left panel: merged DAPI and FITC channels; right panel: FITC panel with cell fluorescence (CF) values calculated for five spermatozoa; fluorescent microscopy, ×1000 magnification.