Confocal reflectance/fluorescence microscopy
We are striving to change how Mohs surgery is conducted, by placing a bedside confocal microscope and developing a rapid staining protocol so that an image equivalent to an H&E stained biopsy can be acquired within 5 min. The Mohs surgeon could stay with the patient and finish multiple excisions of skin cancer, rather than leaving the patient waiting for 30-45 min waiting for histopathology to be done. The staining procedure stains nuclei with one fluorophore (image #1) and cytoplasm with another fluorophore (image #2), and uses reflectance to image collagen fiber bundles (image #3). Then the 3 images are false-colored and combined to yield a single picture that mimics an H&E stained specimen, which is the type of image the Mohs surgeon routinely uses. Figure: Left (c) is false color image of basal cell carcinoma in human patient. Right (d) is H&E stained biopsy (Dan Gareau).
At right is an x-y image of a mouse eye in vivo at a particular depth z within the iris. The image shows dendritic immune cells that have acquired fluorophore-labeled antigen. Fluorescent cells are yellow. Gray image is backscatter of blue light by iris and other soft tissue structures.