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. 2014 Dec 23;1:140046. doi: 10.1038/sdata.2014.46

Figure 2. Functionally guided barrel column extraction.

Figure 2

(a) Chessboard pattern of whisker deprivation. (b) Spared whiskers (magenta) surrounding the C2 whisker (cyan) were stimulated in anesthetized mice. Intrinsic optical signal (IOS) imaged transcranially over left somatosensory cortex (S1). (c) In vivo images of D1-whisker stimulation-evoked IOS peak (pseudocolor) and cerebral surface vasculature (grayscale). (d) Fixed and dissected left S1 cortex with tissue paint and registered IOS peaks (pseudocolor). (e) Remainder tissue after removal of tissue punch centered on the C2 whisker column. (f) Cytochrome oxidase (CO)-stained 80 micron-thick section of remainder tissue registered to the intact remainder tissue, with barrel field traced to confirm the correct punch localization (gray overlay). (g) Estimated barrel column positions within embedded tissue punch (yellow outlines) based on vascular and tissue paint features (cf. red traced blood vessels in panel f) with estimate of optimal cross-section through the C1 and C2 barrel columns parallel to the C-row axis (dashed cyan line). (h) Portion of ATomo ribbon imaged for DAPI at 10x magnification. (i) Maximum-intensity z-projection (MIP) of volume reconstruction of 25x magnification images of ribbon in h. Left: DAPI (gray). Right: DAPI (gray) and YFP (green). High-resolution imaging is targeted to C1 & C2, L3-L5a (red outlines). (j) MIP of multi-session volume reconstruction of 63x magnification images of region shown in (i), with YFP (grey) vGluT1 (green) and vGluT2 (red).