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Determination of Mitochondrial Function in Sperm Cells

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Protocols in Semen Biology (Comparing Assays)

Abstract

With the advancement of science, detecting functionality of mitochondria has become one of the desirable parameters to evaluate sperm quality. This is further aided by availability of wide spectral range of fluorescent probes that has advantage of simultaneous multi-parametric assays. Apart from application of fluorescent probes, computer-assisted image analyzers can be used to assess mitochondrial functionality via motility attributes. In this chapter, we have attempted to describe evaluation of mitochondrial functional status using fluorescent dyes and thus have listed relative merits, protocols, and what to look for in the stained sperm sample. This important chapter also includes triple staining of sperm cells to elucidate integrity of acrosome, plasma membrane, and mitochondrial functions simultaneously. Additionally, procedure for flow cytometry of fluorescent-stained sperm cells as an objective method and tetrazolium (MTT) reduction assay as an easy, inexpensive, and rapid spectrophotometric protocol to determine mitochondrial function in spermatozoa is described.

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Author information

Authors and Affiliations

  1. Quality Control, Semen Freezing Laboratory, ICAR-Central Institute for Research on Cattle, Meerut, 250001, Uttar Pradesh, India

    N. Srivastava

  2. Animal Physiology and Embryo Transfer Laboratory, ICAR-Central Institute for Research on Cattle, Meerut, 250001, Uttar Pradesh, India

    Megha Pande

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  1. N. Srivastava
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  2. Megha Pande
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Correspondence to N. Srivastava .

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Editors and Affiliations

  1. Quality Control, Semen Freezing Laboratory, ICAR-Central Institute for Research on Cattle, Meerut, Uttar Pradesh, India

    N. Srivastava

  2. Animal Physiology and Embryo Transfer Laboratory, ICAR-Central Institute for Research on Cattle, Meerut, Uttar Pradesh, India

    Megha Pande

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Srivastava, N., Pande, M. (2017). Determination of Mitochondrial Function in Sperm Cells. In: Srivastava, N., Pande, M. (eds) Protocols in Semen Biology (Comparing Assays). Springer, Singapore. https://doi.org/10.1007/978-981-10-5200-2_13

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