A correlative method for imaging identical regions of samples by micro-CT, light microscopy, and electron microscopy

Imaging adipose tissue in a model system

Gerhard Sengle, Sara F. Tufa, Lynn Sakai, Martin A. Zulliger, Douglas R. Keene

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

We present a method in which a precise region of interest within an intact organism is spatially mapped in three dimensions by non-invasive micro-computed X-ray tomography (micro-CT), then further evaluated by light microscopy (LM) and transmission electron microscopy (TEM). Tissues are prepared as if for TEM including osmium fixation, which imparts soft tissue contrast in the micro-CT due to its strong X-ray attenuation. This method may therefore be applied to embedded, archived TEM samples. Upon selection of a two-dimensional (2-D) projection from a region of interest (ROI) within the three-dimensional volume, the epoxyembedded sample is oriented for microtomy so that the sectioning plane is aligned with the micro-CT projection. Registration is verified by overlaying LM images with 2-D micro-CT projections. Structures that are poorly resolved in the micro-CT may be evaluated at TEM resolution by observing the next serial ultrathin section, thereby accessing the same ROI by all three imaging techniques. We compare white adipose tissue within the forelimbs of mice harboring a lipid-altering mutation with their littermate controls. We demonstrate that individual osmium-stained lipid droplets as small as 15 μm and separated by as little as 35 μm may be discerned as separate entities in the micro-CT, validating this to be a high-resolution, non-destructive technique for evaluation of fat content.

Original languageEnglish (US)
Pages (from-to)263-271
Number of pages9
JournalJournal of Histochemistry and Cytochemistry
Volume61
Issue number4
DOIs
StatePublished - Jul 3 2013

Fingerprint

X-Ray Microtomography
Adipose Tissue
Microscopy
Electron Microscopy
Transmission Electron Microscopy
Light
Osmium
Microtomy
White Adipose Tissue
Forelimb
Fats
X-Rays
Lipids
Mutation

Keywords

  • Adipose tissue
  • Correlative microscopy
  • Electron microscopy
  • Fat
  • Light microscopy
  • Micro-CT

ASJC Scopus subject areas

  • Anatomy
  • Histology

Cite this

A correlative method for imaging identical regions of samples by micro-CT, light microscopy, and electron microscopy : Imaging adipose tissue in a model system. / Sengle, Gerhard; Tufa, Sara F.; Sakai, Lynn; Zulliger, Martin A.; Keene, Douglas R.

In: Journal of Histochemistry and Cytochemistry, Vol. 61, No. 4, 03.07.2013, p. 263-271.

Research output: Contribution to journalArticle

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