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Revisiting rat spermatogenesis with MALDI imaging at 20- μm resolution.
Mol. Cell. Proteomics 10:M110.005991 (2011)
Matrix-assisted laser desorption/ionization (MALDI) molecular imaging technology attracts increasing attention in the field of biomarker discovery. The unambiguous correlation between histopathology and MALDI images is a key feature for success. MALDI imaging mass spectrometry (IMS) at high definition thus calls for technological developments that were established by a number of small steps. This included tissue and matrix preparation steps, dedicated lasers for MALDI imaging, increase of the robustness against cell debris and matrix sublimation, software for precision matching of molecular and microscopic images, and the analysis of MALDI imaging data using multivariate statistical methods. The goal of these developments is to approach single cell resolution with IMS. Currently a performance level of 20 µm image resolution was achieved with an unmodified and commercially available instrument for proteins detected in the 2-16 kDa range. The rat testis was used as a relevant model for validating and optimizing our technological developments. Indeed, testicular anatomy is amongst the most complex found in mammalian bodies. In the present study, we were able to visualize, at 20 µm image resolution level, different stages of germ cell development in testicular seminiferous tubules, to provide a molecular correlate for its well-established stage-specific classification, to identify proteins of interest using a top-down approach and superimpose molecular and immunohistochemistry images.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
Imaging mass spectrometry; spermatogenesis; testis; MALDI; time-of-flight
ISSN (print) / ISBN
1535-9476
e-ISSN
1535-9484
Zeitschrift
Molecular and Cellular Proteomics
Quellenangaben
Band: 10,
Heft: 3,
Artikelnummer: M110.005991
Verlag
American Society for Biochemistry and Molecular Biology
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed