Abstract
This protocol describes a method for registration of in vivo cortical retinotopic map with cytochrome c oxidase (CO) labeled architectonic maps of the same mouse brain through the alignment of vascular fiducials. By recording surface blood vessel pattern and sequential alignment at each step, this method overcomes the challenge imposed by tissue distortion during perfusion, mounting, sectioning and histology procedures. This method can also be generalized to register and align other types of in vivo functional maps like ocular dominance map and spatial/temporal frequency tuning map with various anatomical maps of mouse cortex.
Highlights
[Background] The mouse visual cortex can be segregated into functionally distinct visual areas by in vivo retinotopic mapping (Marshel et al, 2011; Garrett et al, 2014; Zhuang et al, 2017) or by neuronal track-tracing techniques aided by architectonic structures (Olavarria and Montero, 1989; Wang and Burkhalter, 2007)
These different visual areas have distinct response properties and corticocortical connectivity (Andermann et al, 2011; Marshel et al, 2011; Roth et al, 2012; Wang et al, 2011 and 2012). These results suggest that mouse visual areas form segregated visual streams processing different types of visual information (Murakami et al, 2017; Smith et al, 2017)
Studying the mouse visual system in the context of visual area maps is essential to understanding the organization of visual cortex
Summary
Record vasculature structure image of the cranial window via a fluorescence or a brightfield image (using a green wavelength may give better image contrast of blood vessels). Generate in vivo retinotopic maps through the cranial window (i.e., intrinsic signal Juavinett et al, 2017 or fluorescence retinotopic map Zhuang et al, 2017). Since the animal was perfused by 1% PFA, the brain tissue will be relatively soft for cortex flattening.
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