Journal
APPLIED SURFACE SCIENCE
Volume 252, Issue 19, Pages 6975-6981Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.apsusc.2006.02.197
Keywords
cryofixation; imaging mass spectrometry; high-pressure freezing; Bi-LMIG
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Time-of-flight secondary ion mass spectrometry (TOF-SIMS) was utilized to address the issue of cholesterol localization in rat cerebellum, a subject not previously investigated. Rat cerebellum was prepared by three different procedures: (1) fixation in formaldehyde, freeze-protection by sucrose, freezing in liquid nitrogen and sectioning by cryoultramicrotomy and drying at room temperature or (2) freezing in liquid nitrogen, cryostat sectioning at -40 degrees C and drying at room temperature or (3) high-pressure freezing, freeze-fracturing and freeze-drying. The samples were analyzed in an imaging TOF-SIMS instrument equipped with a Bi-1-7(+)-source. The cholesterol signal (m/z 369 and 385), showed high intensity in the glial cells in white matter and lower intensity in Purkinje cells and in nuclei of granular layer cells. Specimen treated by procedure I showed some signs of diffusion of cholesterol in the tissue. Specimen treated by procedure 2 showed freeze-damage of the cells. Specimen treated by procedure 3 showed distinct localization of cholesterol in well preserved tissue. Thus, high-pressure freezing and freeze-fracturing was used for further characterization of the distribution of cholesterol in rat cerebellum. (c) 2006 Elsevier B.V. All rights reserved.
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