According to the manufacturer’s instructions, the SpermBlue staining method (SpermBlue®, Microptic S.L., Barcelona, Spain) was applied for the assessment of sperm morphology. For each semen sample, 200 spermatozoa were counted microscopically (×400; Zeiss, Oberkochen, Germany) and the results were described as percentage of spermatozoa with normal morphology or with morphological abnormalities including head and integrity of acrosome membrane, midpiece, tail, and cytoplasmic droplets.
Spermblue staining method
Manufactured by Microptic
Sourced in Spain
SpermBlue is a staining method used to facilitate the microscopic examination and analysis of sperm cells. It provides clear visualization of sperm morphology and enables the identification of various sperm parameters, such as concentration, motility, and morphology. The SpermBlue staining method is a tool used in the field of andrology and assisted reproductive technology.
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Sperm Morphology Assessment Using SpermBlue
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Sperm Morphology Assessment Using SpermBlue
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The sperm morphology was assessed using SpermBlue staining method (Microptic SL, Barcelona, Spain) according to Van Der Horst & Maree (2009) (link). Pipetted 10 μl of sperm sample on the edge of the slide and drag the drop with a second slide following 45° angle. Waited for 20–30 s at room temperature. Dropped the slide into deionized water twice. Located the slide on a vertical position and left it to air dry at room temperature. Stained slides were used to perform morphology evaluation using the morphometry module of the Sperm Class Analyzer (Microptic SL, Barcelona, Spain). A total of 200 sperms were analyzed (Ilgin et al., 2017 (link)).
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