Systems and methods for in-operating-theatre imaging of fresh tissue resected during surgery for pathology assessment

A system for in-operating-theatre imaging of fresh tissue samples resected during surgery for pathology assessment, the system comprising:

• a light source for providing an illumination beam that illuminates a fluorescent stained, fresh sample, wherein the fresh sample is held by a sample holder located in an operating theatre;
• a beam expander for expanding a waist of the illumination beam to a size comparable to a field of view to be illuminated, thereby providing a collimated illumination beam;
• a beam splitter located between the sample and a detector array, for directing the collimated illumination beam toward a micro optical element array;
• the micro optical element array comprising a plurality of micro optical elements, wherein each micro optical element of the micro optical element array:
• (i) collects a first portion of the collimated illumination beam and focuses an entirety of that portion onto a single focal point, thereby forming a focal plane comprising an array of tight foci for exciting fluorescence in the sample to produce back-emitted light, and
• (ii) collects back-emitted light from the sample, and the collected back-emitted light propagates and is directed to the detector array;
• a transparent window onto which the sample is placed for imaging, wherein a gap of less than 500 μm is maintained between the micro optical element array and the transparent window, and wherein the focal plane is located at a distance from 10 μm to 200 μm above the transparent window;
• a high precision scanning stage for laterally moving a position of the micro optical element array relative to the transparent window and the detector array along a scan pattern during a single scan such that back-emitted light collected by the micro optical element array is detected by the detector array to form a scanned confocal image corresponding to an area scanned by the scan pattern during the single scan wherein:
• a position of the transparent window relative to the detector array is fixed and
• the scanning stage and micro optical element array are confined within the system such that the scanning stage and micro optical element array are protected from the sample by the transparent window;
• an aperture stop for spatially filtering the back-emitted light collected by the micro optical element array, thereby rejecting out-of-focus light;
• the detector array, wherein the detector array comprises a plurality of detectors, each detector independently detecting a portion of the back-emitted light collected by a micro optical element in the micro optical element array;
• and
• a computing device comprising a processor and a memory storing instructions thereon that, when executed by the processor, cause the processor to construct an image representing an optical slice of the fresh tissue sample based on the back-emitted light detected by the detector array.