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Biomed Opt Express. 2017 Jul 10;8(8):3583-3596. doi: 10.1364/BOE.8.003583. eCollection 2017 Aug 1.

Plastic embedding immunolabeled large-volume samples for three-dimensional high-resolution imaging.

Gang Y1,2,3, Liu X1,2,3, Wang X1,2, Zhang Q1,2, Zhou H1,2, Chen R1,2, Liu L1,2, Jia Y1,2, Yin F1,2, Rao G1,2, Chen J4,5, Zeng S1,2.

Author information

1
Britton Chance Center for Biomedical Photonics, School of Engineering Sciences, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074, China.
2
MOE Key Laboratory for Biomedical Photonics, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, Hubei 430074, China.
3
These authors contributed equally to this work.
4
Department of Cell Biology and Program in Molecular Cell Biology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China.
5
Second Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang 310058, China.

Abstract

High-resolution three-dimensional biomolecule distribution information of large samples is essential to understanding their biological structure and function. Here, we proposed a method combining large sample resin embedding with iDISCO immunofluorescence staining to acquire the profile of biomolecules with high spatial resolution. We evaluated the compatibility of plastic embedding with an iDISCO staining technique and found that the fluorophores and the neuronal fine structures could be well preserved in the Lowicryl HM20 resin, and that numerous antibodies and fluorescent tracers worked well upon Lowicryl HM20 resin embedding. Further, using fluorescence Micro-Optical sectioning tomography (fMOST) technology combined with ultra-thin slicing and imaging, we were able to image the immunolabeled large-volume tissues with high resolution.

KEYWORDS:

(160.2540) Fluorescent and luminescent materials; (170.3880) Medical and biological imaging; (180.2520) Fluorescence microscopy

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