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Cell. 2014 Nov 6;159(4):896-910. doi: 10.1016/j.cell.2014.10.010. Epub 2014 Oct 30.

iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging.

Author information

1
Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
2
Bio-Imaging Resource Center, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
3
Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA. Electronic address: marctl@rockefeller.edu.

Abstract

The visualization of molecularly labeled structures within large intact tissues in three dimensions is an area of intense focus. We describe a simple, rapid, and inexpensive method, iDISCO, that permits whole-mount immunolabeling with volume imaging of large cleared samples ranging from perinatal mouse embryos to adult organs, such as brains or kidneys. iDISCO is modeled on classical histology techniques, facilitating translation of section staining assays to intact tissues, as evidenced by compatibility with 28 antibodies to both endogenous antigens and transgenic reporters like GFP. When applied to degenerating neurons, iDISCO revealed unexpected variability in number of apoptotic neurons within individual sensory ganglia despite tight control of total number in all ganglia. It also permitted imaging of single degenerating axons in adult brain and the first visualization of cleaved Caspase-3 in degenerating embryonic sensory axons in vivo, even single axons. iDISCO enables facile volume imaging of immunolabeled structures in complex tissues. PAPERCLIP.

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PMID:
25417164
DOI:
10.1016/j.cell.2014.10.010
[Indexed for MEDLINE]
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