Interferometric phase microscopy for label free morphological evaluation of sperm cells

Sperm imaging is typically performed using label-based bright-field microscopy, which cannot be used for intracytoplasmic sperm injection. Interferometric phase microscopy was used to evaluate sperm cell morphology without labeling.

Miki Haifler, M.D., Pinhas Girshovitz, M.Sc., Gili Band, Ph.D., Gili Dardikman, Igal Madjar, M.D., Natan T. Shaked, Ph.D.

Volume 104, Issue 1, Pages 43–47


To compare label-free interferometric phase microscopy (IPM) to label-free and label-based bright-field microscopy (BFM) in evaluating sperm cell morphology. This comparison helps in evaluating the potential of IPM for clinical sperm analysis without staining.

Comparison of imaging modalities.

University laboratory.

Sperm samples were obtained from healthy sperm donors.

We evaluated 350 sperm cells, using portable IPM and BFM, according to World Health Organization (WHO) criteria. The parameters evaluated were length and width of the sperm head and midpiece; size and width of the acrosome; head, midpiece, and tail configuration; and general normality of the cell.

Main Outcome Measure(s):
Continuous variables were compared using the Student’s t test. Categorical variables were compared with the χ2 test of independence. Sensitivity and specificity of IPM and label-free BFM were calculated and compared with label-based BFM.

No statistical differences were found between IPM and label-based BFM in the WHO criteria. In contrast, IPM measurements of head and midpiece width and acrosome area were different from those of label-free BFM. Sensitivity and specificity of IPM were higher than those of label-free BFM for the WHO criteria.

Label-free IPM can identify sperm cell abnormalities, with an excellent correlation with label-based BFM, and with higher accuracy compared with label-free BFM. Further prospective clinical trials are required to enable IPM as part of clinical sperm selection procedures.

  • msamplaski

    The selection of the
    “best” sperm for IVF/ICSI is clearly still a relatively rudimentary (and
    imperfect) process. IPM may be a novel tool for identifying the most normal
    appearing sperm. While this is an early study, it is technology worth
    investigating. How common is the equipment for this technique? Is it found in
    most pathology labs? If not, what is the approximate cost? This will obviously
    impact the widespread use of any new equipment and technique.

  • ranjithrama

    Great tool to evaluate sperm morphology without staining. Can the technology be used on fresh semen samples without fixing so sperm selected using this technology can then be used for ICSI? Can this technology be used in sperm identification within testicular tissue?

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