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Optimization of Whole-Body Zebrafish Sectioning Methods for Mass Spectrometry Imaging
Nelson, K. A., G. J. Daniels, J. W. Fournie, AND M. J. Hemmer. Optimization of Whole-Body Zebrafish Sectioning Methods for Mass Spectrometry Imaging. Journal of Biomolecular Techniques. The Association of Biomolecular Resource Facilities, Santa Fe, NM, 24(3):119-127, (2013).
Mass spectrometry imaging methods and protocols have become widely adapted to a variety of tissues and species. However, the mass spectrometry imaging literature contains minimal information on whole-body cryosection preparation for the zebrafish (Danio rerio), a model organism routinely used in developmental, toxicity, and carcinogenicity studies. The optimal medium for embedding and cryosectioning a whole organism or soft tissue specimen for histological examination is a synthetic polymer mixture that is incompatible with mass spectrometry imaging due to ion suppression. We describe the optimal methods and results for embedding and cryosectioning whole-body zebrafish for mass spectrometry imaging. We evaluated 11 embedding media formulations and found a supportive hydrogel with the consistency of cartilage to be the optimal embedding medium. The hydrogel medium does not interfere with mass spectrometry imaging data collection, aids in tissue stability, is readily available for purchase, and is easy to prepare and handle during cryosectioning. Additionally, we decreased the matrix cluster interference commonly caused by alpha-cyano-4-hydroxycinnamic acid by adding ammonium phosphate to the solvent spray solution. The optimized methods developed in our laboratory produced high quality cryosections as well as high quality mass spectral images of sectioned zebrafish.
We describe the optimal methods and results for embedding and cryosectioning whole-body zebrafish for mass spectrometry imaging.
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Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL HEALTH AND ENVIRONMENTAL EFFECTS RESEARCH LAB
GULF ECOLOGY DIVISION