光学透明技术在植物多尺度成像中的应用

doi:10.11983/CBB21175

植物学报 ›› 2022, Vol. 57 ›› Issue (1): 98-110.DOI: 10.11983/CBB21175

光学透明技术在植物多尺度成像中的应用

马灵玉¹^,^3†, 祁晓红^1†, 胡子建¹^,², 沈微微¹^,², 王广超¹^,², 张柏林¹^,²^,⁴, 张曦¹^,²^,⁴^,^*(), 林金星¹^,²^,⁴^,^*()

¹北京林业大学生物科学与技术学院, 北京 100083
²北京林业大学树木与基因编辑研究院, 北京 100083
³中国林业科学研究院木材工业研究所, 北京 100091
⁴北京林业大学生物科学与技术学院, 林木育种国家工程实验室, 北京 100083

收稿日期:2021-10-11 接受日期:2022-01-13 出版日期:2022-01-01 发布日期:2022-01-17
通讯作者: 张曦,林金星
作者简介:linjx@ibcas.ac.cn
* E-mail: zhangxi@bjfu.edu.cn;
第一联系人：
^†共同第一作者
基金资助:
国家自然科学基金(32000558);国家自然科学基金(32030010)

Applications of Optical Clearing Technique in Multi-scale Imaging

Lingyu Ma¹^,^3†, Xiaohong Qi^1†, Zijian Hu¹^,², Weiwei Shen¹^,², Guangchao Wang¹^,², Bolin Zhang¹^,²^,⁴, Xi Zhang¹^,²^,⁴^,^*(), Jinxing Lin¹^,²^,⁴^,^*()

¹College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China
²Institute of Tree and Genome Editing, Beijing Forestry University, Beijing 100083, China
³Research Institute of Wood Industry, Chinese Academy of Forestry, Beijing 100091, China
⁴National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China

Received:2021-10-11 Accepted:2022-01-13 Online:2022-01-01 Published:2022-01-17
Contact: Xi Zhang,Jinxing Lin
About author:First author contact:
^†These authors contributed equally to this paper

摘要/Abstract

摘要： 光学透明技术是一种通过各种化学试剂, 将原本不透明的生物样本实现透明化, 并在光学显微镜下深度成像的技术。结合多种光学显微成像新技术, 光学透明技术可对整个组织进行成像和三维重建, 深度剖析生物体内部空间特征与形成机制。近年来, 多种植物光学透明技术和多尺度成像技术被陆续研发, 并取得了丰硕的研究成果。该文综述了生物体光学透明技术的基本原理和一些新技术, 重点介绍基于光学透明技术开发的新型成像方法及其在植物成像与细胞生物学中的应用, 为后续植物整体、组织或器官的透明、成像与三维重构及功能研究提供理论依据和技术支持。

关键词: 植物整体透明, 多尺度成像, 光学透明技术, 三维重构

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

马灵玉, 祁晓红, 胡子建, 沈微微, 王广超, 张柏林, 张曦, 林金星. 光学透明技术在植物多尺度成像中的应用. 植物学报, 2022, 57(1): 98-110.

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. Chinese Bulletin of Botany, 2022, 57(1): 98-110.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB21175

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

图/表 2

图1 植物样本光学透明、三维(3D)成像和数据分析流程 (A) 植物样本的标记与光学透明, 植物样本的标记主要分为遗传标记(转基因)和组织标记(免疫标记和染料染色); (B) 光学透明后的植物样本3D成像, 可借助光片层荧光显微镜、激光扫描共聚焦显微镜和连续切片断层扫描等方法, 收集连续光学切片的原始图像后, 即可获取目标三维图像信息; (C) 植物样本三维信息统计分析主要包括感兴趣参数的直方图统计、降维分析(UMAP和t-SNE)和多细胞网络拓扑分析

Figure 1 Workflow of optical clearing, three-dimensional (3D) imaging and statistical analysis of plant sample (A) Labeling and optical clearing of plant sample, mainly including genetic labeling (transgene) and histological labeling (immunolabeling and dyeing); (B) 3D imaging of optical cleared plant sample, 3D imaging mainly depends on light-sheet fluorescence microscopy, laser scanning confocal microscopy and serial section tomography, after raw images collection of serial optical sections, 3D images of interest target are displayed; (C) Statistical analysis of plant sample after 3D reconstruction, mainly containing histogram statistics of interest parameters, dimension reduction analysis (uniform manifold approximation and projection (UMAP) and t-distributed stochastic neighbor embedding (t-SNE)) and topological analysis of a multicellular network

表1 不同生物光学透明技术的比较

Table 1 Comparison of clearing techniques in different organisms

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

参考文献 87

[1]	杜中, 赵世绪, 周海鹰, 王莉莉 (1995). 用微分干涉差显微镜和子房整体透明法观察水稻和小麦的胚囊结构. 农业生物技术学报 3(4), 94-97.
[2]	付金娥, 黎颍菁, 韦树根, 潘丽梅, 胡凤云, 童婷婷 (2017). 叶片气孔保卫细胞周长用于鉴定黄花蒿倍性的研究. 北方园艺 (6), 156-161.
[3]	郭海滨, 卢永根, 冯九焕, 杨秉耀, 刘向东 (2006). 利用激光扫描共聚焦显微术对同源四倍体水稻胚囊形成与发育的进一步观察. 激光生物学报 15, 111-117.
[4]	郝建华, 强胜 (2007). 整体透明技术在植物生物学中的应用实例及其剖析. 植物学通报 24, 490-497.
[5]	李国平, 黄群策, 秦广雍 (2006). 用激光扫描共聚焦显微镜观察雪松花粉和花粉管. 激光生物学报 15, 1-8.
[6]	李亚敏, 薛成志, 李贵叶, 胡章立, 阮双琛, 陈玲玲 (2017). 生物组织光学透明技术研究进展. 生物学杂志 34, 83-88.
[7]	李彦坤, 臧巩固, 赵立宁, 李育君, 唐蜻, 程超华 (2011). 整体透明技术观察悬铃叶苎麻胚胎发育的方法研究. 中国麻业科学 33, 142-146.
[8]	王培新, 张丹, 尚爱加, 侯冰 (2016). 组织透明技术. 神经解剖学杂志 32, 124-128.
[9]	谢兆森, 杜鸿儒, 李建宝, Bondada B (2018). 组织透明法观察葡萄叶片生长过程中气孔与叶脉形态结构特征变化. 植物生理学报 54, 237-246.
[10]	杨弘远 (1988). 植物胚胎学中的整体透明技术. 植物学通报 5, 114-116.
[11]	Aoyagi Y, Kawakami R, Osanai H, Hibi T, Nemoto T (2015). A rapid optical clearing protocol using 2,2'-thiodiethanol for microscopic observation of fixed mouse brain. PLoS One 10, e0116280. DOI URL
[12]	Bassel GW (2019). Multicellular systems biology: quantifying cellular patterning and function in plant organs using network science. Mol Plant 12, 731-742. DOI URL
[13]	Becker K, Jährling N, Saghafi S, Weiler R, Dodt HU (2012). Chemical clearing and dehydration of GFP expressing mouse brains. PLoS One 7, e33916. DOI URL
[14]	Belle M, Godefroy D, Couly G, Malone SA, Collier F, Giacobini P, Chedotal A (2017). Tridimensional visualization and analysis of early human development. Cell 169, 161-173. DOI URL
[15]	Berson T, von Wangenheim D, Takáč T, Šamajová O, Rosero A, Ovečka M, Komis G, Stelzer EHK, Šamaj J (2014). Trans-Golgi network localized small GTPase RabA1d is involved in cell plate formation and oscillatory root hair growth. BMC Plant Biol 14, 252. DOI URL
[16]	Cahoon CK, Yu ZL, Wang YF, Guo FL, Unruh JR, Slaughter BD, Hawley RS (2017). Superresolution expansion microscopy reveals the three-dimensional organization of the Drosophila synaptonemal complex. Proc Natl Acad Sci USA 114, E6857-E6866.
[17]	Cai RY, Pan CC, Ghasemigharagoz A, Todorov MI, Förstera B, Zhao S, Bhatia HS, Parra-Damas A, Mrowka L, Theodorou D, Rempfler M, Xavier ALR, Kress BT, Benakis C, Steinke H, Liebscher S, Bechmann I, Liesz A, Menze B, Kerschensteiner M, Nedergaard M, Ertürk A (2019). Panoptic imaging of transparent mice reveals whole-body neuronal projections and skull-meninges connections. Nat Neurosci 22, 317-327. DOI URL
[18]	Chung K, Deisseroth K (2013). CLARITY for mapping the nervous system. Nat Methods 10, 508-513. DOI URL
[19]	Costantini I, Ghobril JP, Di Giovanna AP, Mascaro ALA, Silvestri L, Müllenbroich MC, Onofri L, Conti V, Vanzi F, Sacconi L, Guerrini R, Markram H, Iannello G, Pavone FS (2015). A versatile clearing agent for multi-modal brain imaging. Sci Rep 5, 9808. DOI PMID
[20]	Dodt HU, Leischner U, Schierloh A, Jährling N, Mauch CP, Deininger K, Deussing JM, Eder M, Zieglgänsberger W, Becker K (2007). Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat Methods 4, 331-336.
[21]	Duran-Nebreda S, Bassel GW (2017). Bridging scales in plant biology using network science. Trends Plant Sci 22, 1001-1003. DOI PMID
[22]	Ertürk A, Bradke F (2013). High-resolution imaging of entire organs by 3-dimensional imaging of solvent cleared organs (3DISCO). Exp Neurol 242, 57-64. DOI URL
[23]	Freifeld L, Odstrcil I, Förster D, Ramirez A, Gagnon JA, Randlett O, Costa EK, Asano S, Celiker OT, Gao RX, Martin-Alarcon DA, Reginato P, Dick C, Chen LL, Schoppik D, Engert F, Baier H, Boyden ES (2017). Expansion microscopy of zebrafish for neuroscience and developmental biology studies. Proc Natl Acad Sci USA 114, E10799-E10808.
[24]	Gardner RO (1975). An overview of botanical clearing technique. Stain Technol 50, 99-105. PMID
[25]	Gualda E, Moreno N, Tomancak P, Martins GG (2014). Going ‘open' with Mesoscopy: a new dimension on multi- view imaging. Protoplasma 251, 363-372. DOI URL
[26]	Hama H, Hioki H, Namiki K, Hoshida T, Kurokawa H, Ishidate F, Kaneko T, Akagi T, Saito T, Saido T, Miyawaki A (2015). ScaleS: an optical clearing palette for biological imaging. Nat Neurosci 18, 1518-1529.
[27]	Hama H, Kurokawa H, Kawano H, Ando R, Shimogori T, Noda H, Fukami K, Sakaue-Sawano A, Miyawaki A (2011). Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. Nat Neurosci 14, 1481-1488. DOI URL
[28]	Hasegawa J, Sakamoto Y, Nakagami S, Aida M, Sawa S, Matsunaga S (2016). Three-dimensional imaging of plant organs using a simple and rapid transparency technique. Plant Cell Physiol 57, 462-472. DOI PMID
[29]	Herr JM (1971). A new clearing-squash technique for the study of ovule development in angiosperms. Am J Bot 58, 785-790. DOI URL
[30]	Horecker BL (1943). The absorption spectra of hemoglobin and its derivatives in the visible and near infra-red regions. J Biol Chem 148, 173-183. DOI URL
[31]	Hou B, Zhang D, Zhao S, Wei MP, Yang ZF, Wang SX, Wang JR, Zhang X, Liu B, Fan LZ, Li Y, Qiu ZL, Zhang C, Jiang TZ (2015). Scalable and DiI-compatible optical clearance of the mammalian brain. Front Neuroanat 9, 19.
[32]	Jackson MDB, Xu H, Duran-Nebreda S, Stamm P, Bassel GW (2017). Topological analysis of multicellular complexity in the plant hypocotyl. eLife 6, e26023. DOI URL
[33]	Jing D, Yi Y, Luo W, Zhang S, Yuan Q, Wang J, Lachika E, Zhao Z, Zhao H (2019). Tissue clearing and its application to bone and dental tissues. J Dent Res 98, 621-631. DOI PMID
[34]	Jing D, Zhang SW, Luo WJ, Gao XF, Men Y, Ma C, Liu XH, Yi YT, Bugde A, Zhou BO, Zhao ZH, Yuan Q, Feng JQ, Gao L, Ge WP, Zhao H (2018). Tissue clearing of both hard and soft tissue organs with the PEGASOS method. Cell Res 28, 803-818. DOI URL
[35]	Jo Y, Cho H, Park WS, Kim G, Ryu D, Kim YS, Lee M, Park S, Lee MJ, Joo H, Jo H, Lee S, Lee S, Min HS, Heo WD, Park Y (2021). Label-free multiplexed microtomography of endogenous subcellular dynamics using generalizable deep learning. Nat Cell Biol 23, 1329-1337. DOI URL
[36]	Johnsen S, Widder EA (1999). The physical basis of transparency in biological tissue: ultrastructure and the minimization of light scattering. J Theor Biol 199, 181-198. PMID
[37]	Kao P, Nodine MD (2019). Transcriptional activation of Arabidopsis zygotes is required for initial cell divisions. Sci Rep 9, 17159. DOI URL
[38]	Ke MT, Fujimoto S, Imai T (2013). SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction. Nat Neurosci 16, 1154-1161. DOI URL
[39]	Ke MT, Nakai Y, Fujimoto S, Takayama R, Yoshida S, Kitajima TS, Sato M, Imai T (2016). Super-resolution mapping of neuronal circuitry with an index-optimized clearing agent. Cell Rep 14, 2718-2732. DOI URL
[40]	Kenrick J, Kaul V, Williams EG (1986). Self-incompatibility in Acacia retinodes: site of pollen-tube arrest is the nucellus. Planta 169, 245-250. DOI PMID
[41]	Kierzkowski D, Routier-Kierzkowska AL (2019). Cellular basis of growth in plants: geometry matters. Curr Opin Plant Biol 47, 56-63. DOI PMID
[42]	Klingberg A, Hasenberg A, Ludwig-Portugall I, Medyukhina A, Männ L, Brenzel A, Engel DR, Figge MT, Kurts C, Gunzer M (2017). Fully automated evaluation of total glomerular number and capillary tuft size in nephritic kidneys using lightsheet microscopy. J Am Soc Nephrol 28, 452-459. DOI PMID
[43]	Kubalová I, Černohorská MS, Huranová M, Weisshart K, Houben A, Schubert V (2020). Prospects and limitations of expansion microscopy in chromatin ultrastructure determination. Chromosome Res 28, 355-368. DOI URL
[44]	Kubota SI, Takahashi K, Nishida J, Morishita Y, Ehata S, Tainaka K, Miyazono K, Ueda HR (2017). Whole-body profiling of cancer metastasis with single-cell resolution. Cell Rep 20, 236-250. DOI URL
[45]	Kurihara D, Mizuta Y, Sato Y, Higashiyama T (2015). ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging. Development 142, 4168-4179. DOI PMID
[46]	Kuwajima T, Sitko AA, Bhansali P, Jurgens C, Guido W, Mason C (2013). Clear^T: a detergent-and solvent-free clearing method for neuronal and non-neuronal tissue. Development 140, 1364-1368. DOI PMID
[47]	Lucas M, Kenobi K, von Wangenheim D, Voβ U, Swarup K, De Smet I, Van Damme D, Lawrence T, Péret B, Moscardi E, Barbeau D, Godin C, Salt D, Guyomarc'h S, Stelzer EHK, Maizel A, Laplaze L, Bennett MJ (2013). Lateral root morphogenesis is dependent on the mechanical properties of the overlaying tissues. Proc Natl Acad Sci USA 110, 5229-5234. DOI URL
[48]	Maizel A, von Wangenheim D, Federici F, Haseloff J, Stelzer EHK (2011). High-resolution live imaging of plant growth in near physiological bright conditions using light sheet fluorescence microscopy. Plant J 68, 377-385. DOI URL
[49]	Mombaerts P, Wang F, Dulac C, Chao SK, Nemes A, Mendelsohn M, Edmondson J, Axel R (1996). Visualizing an olfactory sensory map. Cell 87, 675-686. PMID
[50]	Montenegro-Johnson TD, Stamm P, Strauss S, Topham AT, Tsagris M, Wood ATA, Smith RS, Bassel GW (2015). Digital single-cell analysis of plant organ development using 3DCellAtlas. Plant Cell 27, 1018-1033. DOI URL
[51]	Musielak TJ, Slane D, Liebig C, Bayer M (2016). A versatile optical clearing protocol for deep tissue imaging of fluorescent proteins in Arabidopsis thaliana. PLoS One 11, e0161107. DOI URL
[52]	Novák D, Kucharova A, Ovečka M, Komis G, Šamaj J (2016). Developmental nuclear localization and quantification of GFP-tagged EB1c in Arabidopsis root using light-sheet microscopy. Front Plant Sci 6, 1187.
[53]	Ovečka M, Vaškebová L, Komis G, Luptovčiak I, Smertenko A, Šamaj J (2015). Preparation of plants for developmental and cellular imaging by light-sheet microscopy. Nat Protoc 10, 1234-1247. DOI URL
[54]	Ovečka M, von Wangenheim D, Tomančák P, Šamajová O, Komis G, Šamaj J (2018). Multiscale imaging of plant development by light-sheet fluorescence microscopy. Nat Plants 4, 639-650. DOI URL
[55]	Palmer WM, Martin AP, Flynn JR, Reed SL, White RG, Furbank RT, Grof CPL (2015). PEA-CLARITY: 3D molecular imaging of whole plant organs. Sci Rep 5, 13492. DOI URL
[56]	Pan CC, Cai RY, Quacquarelli FP, Ghasemigharagoz A, Lourbopoulos A, Matryba P, Plesnila N, Dichgans M, Hellal F, Ertürk A (2016). Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat Methods 13, 859-867. DOI
[57]	Petersen CCH (2007). The functional organization of the barrel cortex. Neuron 56, 339-355. PMID
[58]	Renier N, Wu ZH, Simon DJ, Yang J, Ariel P, Tessier-Lavigne M (2014). iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging. Cell 159, 896-910. DOI URL
[59]	Richardson DS, Lichtman JW (2015). Clarifying tissue clearing. Cell 162, 246-257. DOI PMID
[60]	Sapala A, Runions A, Smith RS (2019). Mechanics, geometry and genetics of epidermal cell shape regulation: different pieces of the same puzzle. Curr Opin Plant Biol 47, 1-8. DOI URL
[61]	Seo J, Choe M, Kim SY (2016). Clearing and labeling techniques for large-scale biological tissues. Mol Cells 39, 439-446. DOI URL
[62]	Shih CY, Peterson CA, Dumbroff EB (1983). A clearing technique for embryos of lipid-storing seeds. Can J Bot 61, 551-555. DOI URL
[63]	Slattery RA, Grennan AK, Sivaguru M, Sozzani R, Ort DR (2016). Light sheet microscopy reveals more gradual light attenuation in light-green versus dark-green soybean leaves. J Exp Bot 67, 4697-4709. DOI PMID
[64]	Smith BB (1973). The use of a new clearing technique for the study of early ovule development, megasporogenesis, and megagametogenesis in five species of Cornus L. Am J Bot 60, 322-338. DOI URL
[65]	Staudt T, Lang MC, Medda R, Engelhardt J, Hell SW (2007). 2,2'-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy. Microsc Res Tech 70, 1-9. DOI URL
[66]	Susaki EA, Tainaka K, Perrin D, Yukinaga H, Kuno A, Ueda HR (2015). Advanced CUBIC protocols for whole- brain and whole-body clearing and imaging. Nat Protoc 10, 1709-1727. DOI PMID
[67]	Susaki EA, Ueda HR (2016). Whole-body and whole-organ clearing and imaging techniques with single-cell resolution: toward organism-level systems biology in mammals. Cell Chem Biol 23, 137-157. DOI PMID
[68]	Tainaka K, Kubota SI, Suyama TQ, Susaki EA, Perrin D, Ukai-Tadenuma M, Ukai H, Ueda HR (2014). Whole- body imaging with single-cell resolution by tissue decolorization. Cell 159, 911-924. DOI PMID
[69]	Tainaka K, Kuno A, Kubota SI, Murakami T, Ueda HR (2016). Chemical principles in tissue clearing and staining protocols for whole-body cell profiling. Annu Rev Cell Dev Biol 32, 713-741. PMID
[70]	Tainaka K, Murakami TC, Susaki EA, Shimizu C, Saito R, Takahashi K, Hayashi-Takagi A, Sekiya H, Arima Y, Nojima S, Ikemura M, Ushiku T, Shimizu Y, Murakami M, Tanaka KF, Iino M, Kasai H, Sasaoka T, Kobayashi K, Miyazono K, Morii E, Isa T, Fukayama M, Kakita A, Ueda HR (2018). Chemical landscape for tissue clearing based on hydrophilic reagents. Cell Rep 24, 2196-2210. DOI PMID
[71]	Timmers ACJ (2016). Light microscopy of whole plant organs. J Microsc 263, 165-170. DOI URL
[72]	Treweek JB, Chan KY, Flytzanis NC, Yang B, Deverman BE, Greenbaum A, Lignell A, Xiao C, Cai L, Ladinsky MS, Bjorkman PJ, Fowlkes CC, Gradinaru V (2015). Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping. Nat Protoc 10, 1860-1896. DOI URL
[73]	Truckenbrodt S, Sommer C, Rizzoli SO, Danzl JG (2019). A practical guide to optimization in X10 expansion microscopy. Nat Protoc 14, 832-863. DOI PMID
[74]	Truernit E, Bauby H, Dubreucq B, Grandjean O, Runions J, Barthélémy J, Palauqui JC (2008). High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression enables the study of phloem development and structure in Arabidopsis. Plant Cell 20, 1494-1503. DOI URL
[75]	Tuchin VV (2015). Tissue optics and photonics: light-tissue interaction. J Biomed Photonics Eng 1, 98-134.
[76]	Ueda HR, Ertürk A, Chung K, Gradinaru V, Chédotal A, Tomancak P, Keller PJ (2020). Tissue clearing and its applications in neuroscience. Nat Rev Neurosci 21, 61-79. DOI URL
[77]	Vergara HM, Pape C, Meechan KI, Zinchenko V, Genoud C, Wanner AA, Mutemi KN, Titze B, Templin RM, Bertucci PY, Simakov O, Dürichen W, Machado P, Savage EL, Schermelleh L, Schwab Y, Friedrich RW, Kreshuk A, Tischer C, Arendt D (2021). Whole-body integration of gene expression and single-cell morphology. Cell 184, 4819-4837. DOI URL
[78]	Vigouroux RJ, Belle M, Chédotal A (2017). Neuroscience in the third dimension: shedding new light on the brain with tissue clearing. Mol Brain 10, 33. DOI PMID
[79]	von Wangenheim D, Fangerau J, Schmitz A, Smith RS, Leitte H, Stelzer EHK, Maizel A (2016). Rules and self- organizing properties of post-embryonic plant organ cell division patterns. Curr Biol 26, 439-449. DOI PMID
[80]	von Wangenheim D, Hauschild R, Friml J (2017). Light sheet fluorescence microscopy of plant roots growing on the surface of a gel. J Vis Exp (119), e55044.
[81]	Vyplelová P, Ovečka M, Šamaj J (2017). Alfalfa root growth rate correlates with progression of microtubules during mitosis and cytokinesis as revealed by environmental light- sheet microscopy. Front Plant Sci 8, 1870. DOI PMID
[82]	Weissleder R (2001). A clearer vision for in vivo imaging. Nat Biotechnol 19, 316-317. PMID
[83]	Wilkinson LG, Tucker MR (2017). An optimised clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils. Plant Methods 13, 67. DOI PMID
[84]	Wuyts N, Palauqui JC, Conejero G, Verdeil JL, Granier C, Massonnet C (2010). High-contrast three-dimensional imaging of the Arabidopsis leaf enables the analysis of cell dimensions in the epidermis and mesophyll. Plant Methods 6, 17. DOI URL
[85]	Zhang X, Hu ZJ, Guo YY, Shan XY, Li XJ, Lin JX (2020). High-efficiency procedure to characterize, segment, and quantify complex multicellularity in raw micrographs in plants. Plant Methods 16, 100. DOI PMID
[86]	Zhang X, Man Y, Zhuang XH, Shen JB, Zhang Y, Cui YN, Yu M, Xing JJ, Wang GC, Lian N, Hu ZJ, Ma LY, Shen WW, Yang SY, Xu HM, Bian JH, Jing YP, Li XJ, Li RL, Mao TL, Jiao YL, Sodmergen, Ren HY, Lin JX (2021). Plant multiscale networks: charting plant connectivity by multi-level analysis and imaging techniques. Sci China Life Sci 64, 1392-1422. DOI URL
[87]	Zhu MY, Roeder AH (2020). Plants are better engineers: the complexity of plant organ morphogenesis. Curr Opin Genet Dev 63, 16-23. DOI URL

光学透明技术在植物多尺度成像中的应用

Applications of Optical Clearing Technique in Multi-scale Imaging

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 2

参考文献 87

相关文章 0

编辑推荐

Metrics

本文评价