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Optimization of Whole-body Zebrafish Sectioning Methods for Mass Spectrometry Imaging
Citation:
NELSON, K., G. J. DANIELS, J. W. FOURNIE, AND M. J. HEMMER. Optimization of Whole-body Zebrafish Sectioning Methods for Mass Spectrometry Imaging. Presented at Association of Biomolecular Resource Facilities 2012 Conference, Orlando, FL, March 17 - 20, 2012.
Impact/Purpose:
Poster describing new methodology to be presented at annual Association of Biomolecular Resource Facilities
Description:
Mass spectrometry imaging (MSI) methods and protocols have become widely adapted to a variety of tissues and species. However, the MSI literature lacks information on whole-body cryosection preparation for the zebrafish (ZF; Danio rerio), a model organism routinely used in developmental and toxicity studies. The optimal medium for embedding and cryosectioning a whole-body or soft tissue specimen for traditional histological studies is a synthetic polymer mixture that is incompatible with MSI due to peak interference and ion suppression. We describe methods and results for embedding and cryosectioning whole-body ZF from multiple trials to optimize medium compatibility with MSI. A natural polymer commonly used in the food processing industry was determined to be the best embedding medium. The natural polymer medium does not interfere with MSI data collection, aids in tissue stability, is readily available for purchase, and is easy to prepare and handle during cryosectioning. Additionally, we optimized the matrix application step by decreasing the matrix cluster interference commonly caused by alpha-Cyano-4-hydroxycinnamic acid (CHCA) by adding a salt to the solvent spray solution. The optimized methods developed in our laboratory produced high quality cryosections as well as high quality MS images of sectioned ZF.