title: ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging.
abstract: Understanding the structural organization of organs and organisms at the cellular level is a fundamental challenge in biology. This task has been approached by reconstructing three-dimensional structure from images taken from serially sectioned tissues, which is not only labor-intensive and time-consuming but also error-prone. Recent advances in tissue clearing techniques allow visualization of cellular structures and neural networks inside of unsectioned whole tissues or the entire body. However, currently available protocols require long process times. Here, we present the rapid and highly reproducible ACT-PRESTO (active clarity technique-pressure related efficient and stable transfer of macromolecules into organs) method that clears tissues or the whole body within 1 day while preserving tissue architecture and protein-based signals derived from endogenous fluorescent proteins. Moreover, ACT-PRESTO is compatible with conventional immunolabeling methods and expedites antibody penetration into thick specimens by applying pressure. The speed and consistency of this method will allow high-content mapping and analysis of normal and pathological features in intact organs and bodies.
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Eunsoo
authors.initials: E
authors.lastname: Lee
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Jungyoon
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Youhwa
authors.initials: Y
authors.lastname: Jo
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Joo Yeon
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Department of Neural Development and Disease, Korea Brain Research Institute, 701-300 Daegu, Korea.
authors.forename: Yu Jin
authors.initials: YJ
authors.lastname: Jang
authors.affiliation: Department of Neural Development and Disease, Korea Brain Research Institute, 701-300 Daegu, Korea.
authors.forename: Hye Myeong
authors.initials: HM
authors.lastname: Lee
authors.affiliation: Department of Brain &Cognitive Sciences, Graduate School, Daegu Gyeungbuk Institute of Science and Technology (DGIST), Daegu, Korea.
authors.forename: So Yeun
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Logos Biosystems, Inc. Anyang-Si, Gyunggi-Do, 431-755, Republic of Korea.
authors.forename: Ho-Jae
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Logos Biosystems, Inc. Anyang-Si, Gyunggi-Do, 431-755, Republic of Korea.
authors.forename: Keunchang
authors.initials: K
authors.lastname: Cho
authors.affiliation: Logos Biosystems, Inc. Anyang-Si, Gyunggi-Do, 431-755, Republic of Korea.
authors.forename: Neoncheol
authors.initials: N
authors.lastname: Jung
authors.affiliation: Center for Neuroscience, Brain Science Institute, Korea Institute of Science and Technology, Seoul, Korea.|Department of Neuroscience, Korea University of Science and Technology (UST), Daejeon, Korea.
authors.forename: Eun Mi
authors.initials: EM
authors.lastname: Hur
authors.affiliation: Department of Neural Development and Disease, Korea Brain Research Institute, 701-300 Daegu, Korea.
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Jeong
authors.affiliation: Department of Brain &Cognitive Sciences, Graduate School, Daegu Gyeungbuk Institute of Science and Technology (DGIST), Daegu, Korea.
authors.forename: Cheil
authors.initials: C
authors.lastname: Moon
authors.affiliation: Department of Neural Development and Disease, Korea Brain Research Institute, 701-300 Daegu, Korea.
authors.forename: Youngshik
authors.initials: Y
authors.lastname: Choe
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Im Joo
authors.initials: IJ
authors.lastname: Rhyu
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Hyun
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
authors.forename: Woong
authors.initials: W
authors.lastname: Sun
mesh_terms: D020106:Acrylamide / Q000737:chemistry*; D000818:Animals; D001921:Brain / Q000033:anatomy & histology / Q000502:physiology / Q000648:ultrastructure*; D004058:Diffusion; D004586:Electrophoresis; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D016591:Histocytological Preparation Techniques; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D021621:Imaging, Three-Dimensional; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D011108:Polymers / Q000737:chemistry*; D011312:Pressure; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D016707:Tissue Fixation / Q000379:methods; D014981:Xenopus; D015027:Zebrafish
chemical_list: D005404:Fixatives; D020100:Hydrogels; D011108:Polymers; D005557:Formaldehyde; D020106:Acrylamide; C003043:paraform