Figure 3
From: Three-dimensional imaging and quantitative analysis in CLARITY processed breast cancer tissues
The incorporation of the CLARITY process on previously fixed breast cancer core needle biopsy samples preserves the tumor microenvironment. (a) A schematic workflow diagram of tissue processing, clearing, immunostaining, and imaging for the pre-fixed clinical biopsy samples. (b) Gross image of tumor biopsy tissue #8 before vibratome sectioning (2 mm × 1.5 mm), after sectioning (0.5 mm thickness), and post lipid clearing. Representative images from the FFPE corresponding tissue were also H&E stained and displayed (10x, 40x). (c) Confocal microscope imaging of breast cancer tumor #8 (25X). The total 3D volume is visualized in individual and merged channels (left group), and 2D optical section images (25X, right group) demonstrate the staining uniformity that is maintained and increasing imaging depths. The breast tissue was immunostained with antibodies against pan-CK (green), Ki67 (yellow), CD3 (red), and a DAPI nuclear counterstain (blue). Images were visualized by Imaris Version 9.1.2.
