Confocal laser endomicroscope with distal MEMS scanner for real-time histopathology

Confocal laser endomicroscopy is an emerging methodology to perform real time optical biopsy. Fluorescence images with histology-like quality can be collected instantaneously from the epithelium of hollow organs. Currently, scanning is performed at the proximal end of probe-based instruments used routinely in the clinic, and flexibility to control the focus is limited. We demonstrate use of a parametric resonance scanner packaged in the distal end of the endomicroscope to perform high speed lateral deflections. An aperture was etched in the center of the reflector to fold the optical path. This design reduced the dimensions of the instrument to 2.4 mm diameter and 10 mm length, allowing for forward passage through the working channel of a standard medical endoscope. A compact lens assembly provides lateral and axial resolution of 1.1 and 13.6 μm, respectively. A working distance of 0 μm and field-of-view of 250 μm × 250 μm was achieved at frame rates up to 20 Hz. Excitation at 488 nm was delivered to excite fluorescein, an FDA-approved dye, to generate high tissue contrast. The endomicroscope was reprocessed using a clinically-approved sterilization method for 18 cycles without failure. Fluorescence images were collected during routine colonoscopy from normal colonic mucosa, tubular adenomas, hyperplastic polyps, ulcerative colitis, and Crohn's colitis. Individual cells, including colonocytes, goblet cells, and inflammatory cells, could be identified. Mucosal features, such as crypt structures, crypt lumens, and lamina propria, could be distinguished. This instrument has potential to be used as an accessory during routine medical endoscopy.