实验技术与管理2025，Vol.42Issue(3)：9-17,9.DOI:10.16791/j.cnki.sjg.2025.03.002
组织透明化方法对小鼠全脑可视化的影响
Comparison of the effects of different tissue clearing methods on the whole brain visualization of mice
摘要
Abstract
[Objective]Currently,over a hundred tissue clearing methods and their variants exist.Many reports focus on comparing fluorescence signals within specific slices or regions after treatment with several methods.However,these comparisons often cover a limited range of clearing methods and provide one-sided evaluations,making it challenging for researchers to choose the best technique for whole brain visualization studies.[Methods]This study aims to explore the best clearing scheme for mouse tissues expressing endogenous fluorescent proteins;it evaluates the effects of different tissue clearing methods such as CLARITY,CUBIC,FRUIT,3DISCO,uDISCO,and PEGASOS on visualization parameters.These parameters include the transparency degree of the whole mouse brain,the retention of endogenous fluorescence intensity,the count of neuronal somata,and changes in brain volume after clearing.[Results]The analysis indicated that the CLARITY method achieved the highest degree of whole mouse brain clearing;it also excelled in retaining endogenous fluorescence intensity and enabled accurate neuron cell body counts.However,it caused the volume of the whole brain to expand,requiring specialized imaging equipment to accommodate the enlarged sample.On the contrary,the CUBIC and FRUIT methods exhibited relatively poor clearing degrees.The retention intensity of endogenous fluorescence was weak,and neuron cell body count was also less satisfactory.Nevertheless,these methods maintain the volume of the whole brain.Moreover,their cost and technical threshold were low,making them suitable for beginners in tissue clearing research.These novices could start their experiments with relative ease and affordability.The 3DISCO,uDISCO,and PEGASOS methods,on the other hand,all demonstrated a high degree of clearing and significantly reduced the volume of the whole brain.This reduction offered researchers more flexibility in choosing an imaging device.However,both 3DISCO and uDISCO methods were less effective in retaining endogenous fluorescence and counting neuron cell bodies.As a result,they were not very suitable for mouse tissues expressing endogenous fluorescent proteins.By contrast,the PEGASOS method provided a strong balance of performance,retaining fluorescence intensity and supporting accurate neuron cell body counts while also reducing brain volume.Considering all aspects,it emerged as the preferred method for visualizing the whole brain and clearing mouse tissues expressing endogenous fluorescent proteins.It offered a balanced solution that met the requirements of high-quality visualization and practical application in the study of these specific mouse tissues.[Conclusions]This research assists whole brain visualization researchers in selecting appropriate clearing methods based on critical factors such as fluorescence intensity retention,neuron cell body preservation,and sample volume change.关键词
组织透明化方法/透明程度/内源荧光保留强度/神经元胞体计数/全脑体积Key words
tissue clearing method/transparency degree/retention intensity of endogenous fluorescence/neuronal somata count/whole mouse brain volume分类
医药卫生引用本文复制引用
张丹,连佳欢,冯倩倩..组织透明化方法对小鼠全脑可视化的影响[J].实验技术与管理,2025,42(3):9-17,9.基金项目
清华大学实验室创新基金(500-100006029,500-10130218) （500-100006029,500-10130218）
