Fig. 3: High-throughput whole-organ deep immunostaining with dense mapping using INSIHGT.

a Parallelized sample processing with INSIHGT, exemplified with whole mouse kidneys. b Whole organ imaging results with parallelized INSIHGT for the samples shown in (a), showing vimentin INSIHGT signals color-coded by z-depth. One sample was dropped due to manual errors. c,d Whole mouse kidney densely multiplexed visualization with Lycopersicon esculentum lectin (red), Peanut agglutinin (gray), Griffonia simplicifolia lectin I (blue), and AQP-1 (green). d Enlarged 2D view of the white boxed area in (c). e, f Rendered view of whole mouse brain multiplexed Calbindin, NeuN, and c-Fos mapping of a 3-year-old mouse. g–i Age-related structural and molecular changes in the thalamus (g) and striatum (h) with cavitations (indicated by yellow arrowheads), and the hippocampus (i) with CALB1-positive deposits (indicated by yellow arrows). j Whole brain multiplexed staining of the calcium-binding proteins calbindin (CALB1), calretinin (CALB2), and parvalbumin (PVALB) with 3 days of INSIHGT staining. k Zoomed in 3D rendering view on the hippocampus (Hp), reticular nucleus of thalamus (Ret. Nuc.), and amygdala (Amyg.). l A single coronal slice view (left) and a 2 mm-thick anteroposterior projection (right) of the same sample.