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    Laboratory Analysis of Semen SOP Macros

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    robe_djii
    This document outlines the standard operating procedure for analyzing a semen sample in a laboratory. It describes how to examine the semen macroscopically by assessing appearance, consistency, volume, and pH. Microscopic analysis includes evaluating agglutination, motility, viability, sperm count, and morphological characteristics. Various terminology are defined to classify semen analysis results, such as normospermia, oligospermia, asthenospermia, and teratospermia.

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    Laboratory Analysis of Semen SOP Macros

    Uploaded by

    robe_djii
    611 views3 pages
    This document outlines the standard operating procedure for analyzing a semen sample in a laboratory. It describes how to examine the semen macroscopically by assessing appearance, consistency, volume, and pH. Microscopic analysis includes evaluating agglutination, motility, viability, sperm count, and morphological characteristics. Various terminology are defined to classify semen analysis results, such as normospermia, oligospermia, asthenospermia, and teratospermia.

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    This document outlines the standard operating procedure for analyzing a semen sample in a laboratory. It describes how to examine the semen macroscopically by assessing appearance, consistency, volume, and pH. Microscopic analysis includes evaluating agglutination, motility, viability, sperm count, and morphological characteristics. Various terminology are defined to classify semen analysis results, such as normospermia, oligospermia, asthenospermia, and teratospermia.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    611 views3 pages

    Laboratory Analysis of Semen SOP Macros

    Uploaded by

    robe_djii
    This document outlines the standard operating procedure for analyzing a semen sample in a laboratory. It describes how to examine the semen macroscopically by assessing appearance, consistency, volume, and pH. Microscopic analysis includes evaluating agglutination, motility, viability, sperm count, and morphological characteristics. Various terminology are defined to classify semen analysis results, such as normospermia, oligospermia, asthenospermia, and teratospermia.

    Copyright:

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    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 3

    LABORATORY ANALYSIS OF SEMEN

    SOP

    MACROSCOPY

    APPEARANCE

    Normal semen has a grey-opalescent colour, is homogenous and liquefies within 60minutes at
    room temperature. The sample may appear clear if the sperm concentrate is too low or brown
    when red blood cells are present (haematosperia).

    CONSISTENCY (VISCOSITY)

    It can be estimated by gentle aspiration into a 5mls pipette and then allowing the semen to drop
    by gravity and observing the length of the thread formed.

    VOLUME

    The volume of the ejaculate should be measured either with graduated cylinder or by aspirating
    the whole semen into a wide-mouth pipette.

    PH

    The pH is determined by acidic secretions of the prostate and alkaline secretions of the seminal
    vesicle. It should normally be in the range of 7.2-8.0.

    MICROSCOPY

    AGGLUTINATION

    Agglutination means that mobile spermatozoa stick to each other head to head, middle to middle,
    tail to tail, or mixed. In case of agglutination, semen culture must be performed in order to
    exclude infection with E. coli.

    MOTILITY

    A drop of the semen is delivered onto a microscopic slide and covered with 22X22cm cover slip.
    The microscope field is scanned systematically with X40 objective and a total of 100
    spermatozoa are counted. The motility of the spermatozoa is graded in percentage whether it
    shows

    1. Active linear forward progressive motility


    2. Slow or sluggish progressive motility
    3. Non progressive motility
    4. Non motility

    SPERM VIABILITY

    Vital staining of the spermatozoa allows quantification of the fraction of living cells
    independently of their motility. Live and dead spermatozoa are distinguished by adding one drop
    of 0.5% eosin Y stain to one drop of semen at room temperature and smear the mixture on the
    microscope slide. Using the X40 objective, 100 spermatozoa are classified as either coloured
    orange-red, if the stain has passed through the membrane and therefore the cell is dead, or non-
    stained, the cell being considered alive.

    COUNTING SPERMATOZOA

    In the haemocytomether method, a 1:20 dilution from a well mixed sample is prepared by
    diluting 50µl of liquefied semen with 950µl diluents. The diluents is prepared by adding 50g of
    sodium carbonate, 10ml of 35% (v/v) formalin and making up the solution to a final volume of
    1000ml.

    Using a Pasteur pipette, charge an improved Neubauer chamber solution with well mixed
    solution. Wait 3-5minutes for the spermatozoa to settle. Using the X10 objective, count the
    number of the sperm cells in an area of 2square millimeter (2 large squares). Calculate the
    number of spermatozoa in 1ml of fluid by multiplying the number counted by 100,000

    ANALYSIS OF THE MORPHOLOGICAL CHARACTERISTICS OF SPERMATOZOA

     Make a thin smear of liquefied well-mixed semen on a slide. While wet, fix the smear
    with 95% v/v ethanol for 5-10minutes, and allow to air-dry.
     Wash the slide with sodium bicarbonate-formalin solution to remove any mucus which
    may be present.
     Cover the smear with diluents (1 in 20) carbon fuchsin and allow to stain for 3minutes.
     Counter stain, by covering the smear with dilute (1 in 20) Loeffer methylene blue for
    2minutes
     Wash off the stain with water. Drain and allow the smear to air-dry.

    STAINING RESULTS

     Nucleus of head –Dark blue


     Cytoplasm of head –Pale blue
     Middle piece and tail –pink-red
    TERMINOLOGIES FOR CLASSIFICATION IN SEMEN ANALYSIS

    NORMOSPERMIA Normal sperm count, motility and morphology


    OLIGOZOOSPERMIA Sperm count fewer than 20million/ml
    ASTHENOZOOSPERMIA Fewer than 5% spermatozoa with forward
    progressive movement
    TERATOZOOSPERMIA Fewer than 50% spermatozoa with normal
    morphology
    OLIGOASTHENOTERATOZOOSPERMIA Sperm count, forward progressive movement
    and morphology are disturbed (>50%)
    AZOOSPERMIA No spermatozoa in ejaculate
    ASPERMIA No ejaculate

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