Fig. 5. Compatibility of FLARE with diverse procedures.

FLARE-stained 100 μm thick mouse kidney sections prepared using various procedures that include two different ExM variants, tissue clearing, and the combination with either immunostaining or DNA FISH. (a) 5-color confocal microscopy image of unexpanded mouse kidney tissue that was stained for carbohydrates (green), amines (red), and then immunostained for podocalyxin (magenta), aquaporin-1 (blue), and cytokeratin 8+18 (grey). (b) Confocal microscopy image of unexpanded cleared mouse kidney tissue that was stained for carbohydrates (green), amines (red), and then DNA FISH (blue) against MaSat DNA. (c) Confocal microscopy image of expanded mouse kidney tissue that was treated by the detergent-based protocol and stained for carbohydrates (green), amines (red), and then DNA FISH (blue) against pericentromeric major satellite (MaSat) DNA. (d) Confocal microscopy image of enzyme digested expanded mouse kidney tissue that was stained for carbohydrates (green), amines (red), DNA (blue) and then immunostained for podocalyxin (grey). Scale bars are 50 μm (a, d), 5 μm (b) and 3 μm (c, pre-expansion units). Panels a-b were adapted from Mao et al.³ Science Advances. AAAS Publishing Group.