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Scientific Publication Citing Dragonfly

New insights into sperm ultrastructure through enhanced scanning electron microscopy

Denis Korneev (1), Donna Jo Merriner (1), Gediminas Gervinskas (2), Alex de Marco (2,3), Moira K O’Bryan (1,4)
bioRxiv, November 2020. DOI: 10.1101/2020.11.12.380808


electron microscopy, high-pressure freezing, focused ion beam, male infertility, male fertilitycoustic


The analysis of spermatozoa morphology is fundamental to understand male fertility and the aetiology of infertility. Traditionally scanning electron microscopy (SEM) has been used to define surface topology. Recently, however, it has become a critical tool for three-dimensionally analyse of internal cellular ultrastructure. Modern SEM provides nanometer-scale resolution, but the meaningfulness of such information is proportional to the quality of the sample preservation. In this study, we demonstrate that sperm quickly and robustly adhere to gold-coated surfaces. Leveraging this property, we developed three step-by-step protocols fulfilling different needs for sperm imaging: chemically fixed monolayers for SEM examination of the external morphology, and two high-pressure freezing-based protocols for fast SEM examination of full cell internal morphology and focused ion-beam SEM (FIB-SEM) tomography. These analyses allow previously unappreciated insights into mouse sperm ultrastructure, including the identification of novel structures within the fibrous sheath and domain-specific interactions between the plasma membrane and exosome-like structures.

How Our Software Was Used

Dragonfly was used to process and visualize FIB-SEM data stacks.

Author Affiliation

(1) School of Biological Sciences, Monash University, 25 Rainforest Walk, 3800. Victoria, Australia.
(2) Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, 3800 Clayton, Australia.
(3) ARC Centre of Excellence in Advanced Molecular Imaging, Monash University, 3800 Clayton, Australia.
(4) School of BioSciences, Faculty of Science, The University of Melbourne, 3010, Victoria, Australia.

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