Confocal Raman microspectroscopy as an analytical tool to assess the mitochondrial status in human spermatozoa.

Confocal Raman microspectroscopy was used to investigate human sperm cells. Raman mapping with a 532 nm excitation laser allowed to unambiguously characterize the nucleus, the neck, and, in particular, the mitochondria-rich middle piece of a human sperm cell. The effect of ultraviolet radiation on different organelles of the sperm was quantified by localized spectral Raman signatures obtained within milliseconds. Chemical changes within the sub-cellular structure of the sperm cells were recorded as a function of ultraviolet light exposure time, showing the proof-of-principle that Raman microspectroscopy can be a fast diagnostic method for detecting the mitochondrial and motility status of human spermatozoa.

[1]  M. Diem,et al.  A decade of vibrational micro-spectroscopy of human cells and tissue (1994-2004). , 2004, The Analyst.

[2]  X. Xie,et al.  Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy Published, JLR Papers in Press, August 16, 2003. DOI 10.1194/jlr.D300022-JLR200 , 2003, Journal of Lipid Research.

[3]  R. Aitken,et al.  A new method for evaluation of the acrosome reaction in viable human spermatozoa , 2009, Andrologia.

[4]  C. Otto,et al.  The intensity of the 1602 cm-1 band in human cells is related to mitochondrial activity , 2009 .

[5]  M. Diem,et al.  Spectroscopy , 2007, Acta Neuropsychiatrica.

[6]  Hiro-o Hamaguchi,et al.  Raman spectroscopic signature of life in a living yeast cell , 2004 .

[7]  G. A. Thomas,et al.  Raman spectra of the model B-DNA oligomer d(CGCGAATTCGCG)2 and of the DNA in living salmon sperm show that both have very similar B-type conformations. , 1986, Biochemistry.

[8]  Weizhong Ke,et al.  Raman spectroscopic study of the influence on herring sperm DNA of heat treatment and ultraviolet radiation , 1999 .

[9]  M. Havenith,et al.  Fast quantification of water in single living cells by near-infrared microscopy. , 2004, The Analyst.

[10]  P. Lishko,et al.  Acid Extrusion from Human Spermatozoa Is Mediated by Flagellar Voltage-Gated Proton Channel , 2010, Cell.

[11]  Andreas Volkmer,et al.  Vibrational Imaging with High Sensitivity via Epidetected Coherent Anti-Stokes Raman Scattering Microscopy , 2001 .

[12]  H. Hamaguchi,et al.  Behaviors of the “Raman Spectroscopic Signature of Life” in Single Living Fission Yeast Cells under Different Nutrient, Stress, and Atmospheric Conditions , 2007, Applied spectroscopy.

[13]  Abdelilah Beljebbar,et al.  Raman spectral imaging of single living cancer cells: a preliminary study. , 2009, The Analyst.

[14]  D.M. Mount,et al.  An Efficient k-Means Clustering Algorithm: Analysis and Implementation , 2002, IEEE Trans. Pattern Anal. Mach. Intell..

[15]  R. Turner.  Tales from the tail: what do we really know about sperm motility? , 2003, Journal of andrology.

[16]  L. Zamboni.  Physiology and pathophysiology of the human spermatozoon: the role of electron microscopy. , 1991, Journal of electron microscopy technique.

[17]  X. Xie,et al.  Coherent Anti-Stokes Raman Scattering Microscopy: Instrumentation, Theory, and Applications , 2004 .

[18]  Christoph Krafft,et al.  Identification of organelles and vesicles in single cells by Raman microspectroscopic mapping , 2005 .

[19]  Rod Balhorn,et al.  Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells , 2009, Journal of biophotonics.

[20]  Martina Havenith,et al.  Label-free imaging of metal-carbonyl complexes in live cells by Raman microspectroscopy. , 2010, Angewandte Chemie.

[21]  N. Keiding,et al.  Evidence for decreasing quality of semen during past 50 years. , 1992 .

[22]  Guiwen Wang,et al.  NIR Raman spectroscopic investigation of single mitochondria trapped by optical tweezers. , 2007, Optics express.