Applications of Optical Clearing Technique in Multi-scale Imaging

doi:10.11983/CBB21175

Abstract

Abstract: The optical clearing techniques are common methods that allow biological tissues or organs to be deeply transparent and imaged under an optical microscope by processing with chemical reagents. Driven by a variety of optical microscopy technologies, the entire tissue can be imaged and reconstructed three-dimensionally (3D) using the clearing technique to deeply analyze the internal spatial characteristics and formation mechanism of biological tissues. In recent years, several optical clearing techniques and multi-scale imaging techniques have been developed and fruitful researches have been reported. This article discusses basic principles and some new techniques of optical clearing, with particular focus on new imaging methods based on optical clearing technique and their applications in plant imaging and cell biology. This review will provide theoretical basis and technical support for the subsequent research on clearing, imaging and 3D reconstruction and function of whole plants, tissues or organs.

Key words: clearing of whole plant, multi-scale imaging, optical clearing technique, three-dimensional reconstruction

Lingyu Ma, Xiaohong Qi, Zijian Hu, Weiwei Shen, Guangchao Wang, Bolin Zhang, Xi Zhang, Jinxing Lin. Applications of Optical Clearing Technique in Multi-scale Imaging[J]. Chinese Bulletin of Botany, 2022, 57(1): 98-110.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB21175

https://www.chinbullbotany.com/EN/Y2022/V57/I1/98

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References 87

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透明方法	主要成分	作用	优点	缺点	应用	参考文献
3DISCO	四氢呋喃和二氯甲烷	脱水和脱脂	透明效果好, 荧光蛋白保存良好	试剂有毒性, 组织易萎缩	鼠脑, 小鼠脊髓、肺和脾, 免疫器官, 肿瘤组织	Ertu?rk and Bradke, 2013
CUBIC	四乙酸	脱色	操作简单, 透明效率高, 可重复性强	时间较长	小鼠组织	Tainaka et al., 2014; Susaki et al., 2015; Susaki and Ueda, 2016
CLARITY	水凝胶	组织灌注	透明效果好	操作复杂, 组织易褐化	鼠脑	Treweek et al., 2015
TDE	硫二甘醇	RI匹配	简单快速, 组织变形、体积变小	荧光蛋白易淬灭	鼠脑和拟南芥(Arabi- dopsis thaliana)种子	Staudt et al., 2007; Aoyagi et al., 2015; Musielak et al., 2016
强碱	氢氧化钠和氢氧化钾	脱脂和除蛋白	操作简单, 成本低, 透明效率高	组织损伤较大	黄花蒿(Artemisia annua)叶片和葡萄(Vitis vinifera)叶片	付金娥等, 2017; 谢兆森等, 2018
PEA-CLARITY	细胞壁降解酶	水解酶类	透明效果好	操作复杂, 组织易褐化	拟南芥叶片和烟草(Nicotiana tabacum)叶片	Palmer et al., 2015
ClearSee	木糖醇、脱氧胆酸钠和尿素	折射率(RI)匹配	保持荧光蛋白稳定性, 减少叶绿体自发荧光	时间较长	拟南芥幼苗、叶片和雌蕊	Kurihara et al., 2015; Slattery et al., 2016

透明方法	主要成分	作用	优点	缺点	应用	参考文献
3DISCO	四氢呋喃和二氯甲烷	脱水和脱脂	透明效果好, 荧光蛋白保存良好	试剂有毒性, 组织易萎缩	鼠脑, 小鼠脊髓、肺和脾, 免疫器官, 肿瘤组织	Ertu?rk and Bradke, 2013
CUBIC	四乙酸	脱色	操作简单, 透明效率高, 可重复性强	时间较长	小鼠组织	Tainaka et al., 2014; Susaki et al., 2015; Susaki and Ueda, 2016
CLARITY	水凝胶	组织灌注	透明效果好	操作复杂, 组织易褐化	鼠脑	Treweek et al., 2015
TDE	硫二甘醇	RI匹配	简单快速, 组织变形、体积变小	荧光蛋白易淬灭	鼠脑和拟南芥(Arabi- dopsis thaliana)种子	Staudt et al., 2007; Aoyagi et al., 2015; Musielak et al., 2016
强碱	氢氧化钠和氢氧化钾	脱脂和除蛋白	操作简单, 成本低, 透明效率高	组织损伤较大	黄花蒿(Artemisia annua)叶片和葡萄(Vitis vinifera)叶片	付金娥等, 2017; 谢兆森等, 2018
PEA-CLARITY	细胞壁降解酶	水解酶类	透明效果好	操作复杂, 组织易褐化	拟南芥叶片和烟草(Nicotiana tabacum)叶片	Palmer et al., 2015
ClearSee	木糖醇、脱氧胆酸钠和尿素	折射率(RI)匹配	保持荧光蛋白稳定性, 减少叶绿体自发荧光	时间较长	拟南芥幼苗、叶片和雌蕊	Kurihara et al., 2015; Slattery et al., 2016