1Laboratory of Biomedical Physics,Groenenborgerlaan 171,2020 Antwerpen,Belgium.
2Laboratory of Functional Morphology,Universiteitsplein 1,2610 Antwerp,Belgium.
Microsc Microanal. 2014 Aug;20(4):1208-17. doi: 10.1017/S1431927614001329. Epub 2014 Jun 25.
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.
两种方法特别适用于对包含多种组织类型（骨、肌肉、神经或脂肪）的宏观样本进行断层成像和具有组织学细节：光片（基于）荧光显微镜（LSFM）和微计算机断层扫描（micro-CT）。微 CT 需要用重元素化学物质进行染色（因此需要固定，有时还需要脱水），以便在与密度较高的结构一起测量时使软组织成像。LSFM 需要固定、脱钙、脱水、透明和用荧光染料染色。这两种成像方法的标本制备都容易发生收缩，但往往没有提及，更不用说量化了。本文定量确定了所选制备方法/染色剂的收缩存在和程度。LSFM 对骨的体积收缩为 17%，对肌肉的体积收缩为 56%，对脑组织的体积收缩为 62%。三种最流行的 micro-CT 染色剂（磷钨酸、碘化钾和碘在无水乙醇中）对肌肉的体积收缩范围为 10-56%，对脑组织的体积收缩范围为 27-66%，而骨在 micro-CT 制备中不收缩。
