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J Assist Reprod Genet (2011) 28:725729 DOI 10.1007/s10815-011-9584-y
TECHNICAL INNOVATIONS
Digital holographic microscopy in human sperm imaging
Igor Crha & Jana Zakova & Martin Huser &
Pavel Ventruba & Eva Lousova & Michal Pohanka
Received: 29 March 2011 /Accepted: 13 May 2011 /Published online: 11 June 2011 # Springer Science+Business Media, LLC 2011
AbstractPurpose The aim of this study was to use digital holographic microscopy (DHM) in human sperm imaging and compare quantitative phase contrast of sperm heads in normozoospermia (NZ) and oligoasthenozoospermia (OAT).
Methods DHM spermatozoa imaging and repeated quantitative phase shift evaluation were used. Five NZ and 5 OAT samples were examined. Semen samples were examined by semen analysis and processed for DHM. Main outcome measures were maximum phase shift value of the sperm heads. Differences of the phase shift and in NZ and OAT samples were statistically tested. Results In NZ samples median phase shifts were in the range 2.723.21 rad and 2.002.15 in OAT samples. Differences among individual samples were statistically significant (p<0.001) in both groups. Median phase shift according to sperm count was 2.90 rad in NZ samples and2.00 rad in OAT samples. This difference was statistically significant (p<0.001).
Conclusion Quantitative evaluation of the phase shift by DHM could provide new information on the exact structure and composition of the sperm head. At present, this technique is not established for clinical utility.
Keywords Digital holographic microscopy.
Sperm imaging . Spermatozoon . Male infertility. Chromatin integrity
Introduction
Digital holographic microscopy (DHM) is a noninvasive imaging method of quantitative phase contrast. Its main advantage over a conventional Zernike phase contrast is the quantification. This feature enables much more sophisticated evaluation of image data. For this reason it is particularly useful in cell biology for visualization and evaluation of living cells that exhibit phase-objects properties. It is considered to be a very promising tool in single-cell analysis [13]. Following previous experience from study of structural integrity of neoplastic cells [4], we used the DHM in sperm imaging.
The aim of the study was to use DHM in human sperm imaging and compare quantitative phase contrast of sperm heads in normozoospermia and oligoasthenoteratozoospermia.
Materials and methods
Digital holographic microscopy
Holographic microscopy is based on the interference of the light passing through the object with...