Optical Clearing of Fixed Brain Samples Using SeeDB. Issue 1 (15th January 2014)

• Record Type: Journal Article
• Authors: Ke, Meng‐Tsen; Imai, Takeshi
• Editors: Gerfen, Charles R.; Holmes, Andrew; Sibley, David; Skolnick, Phil; Wray, Susan
• Is Part Of: Current protocols in neuroscience. Issue 66:Issue 1(2014)
• Page Start: 2.22.1
• Page End: 2.22.19
• Publication Date: 2014-01-15
• Subjects: optical clearing -- fluorescent imaging -- two‐photon microscopy; Neurosciences -- Laboratory manuals; Neurosciences; Cytological Techniques; Histological Techniques; Nervous System -- anatomy & histology; Neurosciences; Laboratory Manual; Laboratory manuals; 612.8
• DOI: 10.1002/0471142301.ns0222s66
• Languages: English
• ISSNs: 1934-8576

Abstract

Abstract: Large‐scale three‐dimensional fluorescence imaging is essential for comprehensive and quantitative understanding of neuronal circuitry. We describe a water‐based optical clearing agent, SeeDB, which clears fixed brain samples in a few days leaving many types of fluorescent dyes unquenched, including fluorescent proteins and lipophilic neuronal tracers. This method maintains a constant sample volume during the clearing procedure, an important factor to keep cellular morphology intact. After optical clearing with SeeDB, we can reach a depth of >1000 µm with confocal microscopy. When combined with two‐photon microscopy, SeeDB allows us to image fixed mouse brains at millimeter‐scale level. Curr. Protoc. Neurosci . 66:2.22.1‐2.22.19. © 2014 by John Wiley & Sons, Inc.