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Volume Shrinkage of Bone, Brain and Muscle Tissue in Sample Preparation for Micro-CT and Light Sheet Fluorescence Microscopy (LSFM)

Published online by Cambridge University Press:  25 June 2014

Jan Buytaert ,
Jana Goyens ,
Daniel De Greef ,
Peter Aerts  and
Joris Dirckx
Jan Buytaert*
Affiliation:
Laboratory of Biomedical Physics, Groenenborgerlaan 171, 2020 Antwerpen, Belgium
Jana Goyens
Affiliation:
Laboratory of Biomedical Physics, Groenenborgerlaan 171, 2020 Antwerpen, Belgium Laboratory of Functional Morphology, Universiteitsplein 1, 2610 Antwerp, Belgium
Daniel De Greef
Affiliation:
Laboratory of Biomedical Physics, Groenenborgerlaan 171, 2020 Antwerpen, Belgium
Peter Aerts
Affiliation:
Laboratory of Functional Morphology, Universiteitsplein 1, 2610 Antwerp, Belgium
Joris Dirckx
Affiliation:
Laboratory of Biomedical Physics, Groenenborgerlaan 171, 2020 Antwerpen, Belgium
*
*Corresponding author. [email protected]
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Abstract

Two methods are especially suited for tomographic imaging with histological detail of macroscopic samples that consist of multiple tissue types (bone, muscle, nerve or fat): Light sheet (based) fluorescence microscopy (LSFM) and micro-computed tomography (micro-CT). Micro-CT requires staining with heavy chemical elements (and thus fixation and sometimes dehydration) in order to make soft tissue imageable when measured alongside denser structures. LSMF requires fixation, decalcification, dehydration, clearing and staining with a fluorescent dye. The specimen preparation of both imaging methods is prone to shrinkage, which is often not mentioned, let alone quantified. In this paper the presence and degree of shrinkage are quantitatively identified for the selected preparation methods/stains. LSFM delivers a volume shrinkage of 17% for bone, 56% for muscle and 62% for brain tissue. The three most popular micro-CT stains (phosphotungstic acid, iodine with potassium iodide, and iodine in absolute ethanol) deliver a volume shrinkage ranging from 10 to 56% for muscle and 27–66% for brain, while bone does not shrink in micro-CT preparation.

Keywords

stainingshrinkagevolumesoft tissuebonespecimen preparationlight-sheet microscopyX-ray tomography
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 4 , August 2014 , pp. 1208 - 1217
Copyright
© Microscopy Society of America 2014 

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