Figure 2

Pattern recovery at the base of the crypt is accomplished by cells residing in the crypt. (a) Time-lapse images using three-photon microscopy after ablation of a single Lgr5+ ISC in the crypt with Lgr5+ ISC (green) and third harmonic generation (THG, magenta). THG image was taken with 1,300 nm wavelength laser excitation and Lgr5+ GFP image was acquired with 880 nm wavelength laser excitation. Red blood cells produced THG in the vasculature just outside the crypt. Images were overlapped via post-processing. White arrow indicates an ablated cell. Scale bar: 30 µm. (b) Examples of two different crypts showing visualization of Paneth cells with THG and α-lysozyme staining in fixed tissue. THG signal was acquired under three-photon microscopy and α-lysozyme immunofluorescence was imaged under two-photon microscopy. Images were overlapped via post-processing. Scale bar: 30 µm.