Volume Shrinkage of Bone, Brain and Muscle Tissue in Sample Preparation for Micro-CT and Light Sheet Fluorescence Microscopy (LSFM)
Volume Shrinkage of Bone, Brain and Muscle Tissue in Sample Preparation for Micro-CT and Light Sheet Fluorescence Microscopy (LSFM)
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DOI：
10.1017/s1431927614001329
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发表时间：
2014-08-01
影响因子：
2.8
通讯作者：
Dirckx, Joris
中科院分区：
文献类型：
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作者：
Buytaert, Jan;Goyens, Jana;Dirckx, Joris
有两种方法特别适用于包含多种组织类型（骨骼、肌肉、神经或脂肪）的宏观样本的组织学细节的断层成像：基于光片的荧光显微镜（LSFM）和微型计算机断层扫描（micro-CT）。Micro-CT需要用重化学元素染色（因此需要固定，有时需要脱水），以便在测量更致密的结构时使软组织成像。LSMF需要固定、脱钙、脱水、澄清和用荧光染料染色。两种成像方法的标本制备都容易出现收缩，这通常不会被提及，更不用说量化了。在本文中，对所选制备方法/染色剂的收缩存在和程度进行了定量鉴定。LSFM可使骨骼体积收缩17%，肌肉收缩56%，脑组织收缩62%。三种最流行的微CT染色剂（磷钨酸、碘加碘化钾和碘的无水乙醇溶液）可使肌肉体积收缩10 - 56%，大脑体积收缩27-66%，而骨骼在微CT制备中不会收缩。
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.