Sample preparation of animal tissues and cell cultures for secondary ion mass spectrometry (SIMS) microscopy.
Sample preparation is a critical step in the elemental analysis of animal tissues and cell cultures with ion microscopy. Since live cells cannot be analyzed with ion microscopy, a careful sample fixation is necessary which preserves the native structural and chemical integrity of a specimen. The evaluation of morphological and chemical integrity of a fixed specimen is necessary before any physiological explanation of ion fluxes is interpreted based on ion microscopy. For diffusible ion localization studies, strict cryogenic procedures are recommended. Examples are shown for diffusible ion microanalysis in frozen-freeze-dried tissues and cell cultures. Ion microscopy studies of tightly bound elements/molecules may be conducted in chemically fixed and/or plastic embedded specimens. Since it is not generally known which elements/molecules are tightly bound to the tissue matrix, a confirmation of elemental distribution with cryogenic procedures is desirable. A recent approach of combining laser scanning confocal fluorescence microscopy and ion microscopy on the same frozen freeze-dried cell is also discussed for recognizing smaller cytoplasmic structures in ion microscopy images.
Chandra
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Baker Laboratory of Chemistry, Cornell University, Ithaca, NY 14853.
Morrison
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D000818:Animals; D002478:Cells, Cultured / Q000648:ultrastructure; D004058:Diffusion; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D013048:Specimen Handling