A high-resolution MACROscope with differential phase contrast, transmitted light, confocal fluorescence, and hyperspectral capabilities for large-area tissue imaging

Recent advances in imaging technology have contributed greatly to biological science. Confocal fluorescence microscopes can acquire two-dimensional and three-dimensional images of biological samples such as live or fixed cells and tissues. Specimens that are large (e.g., a 10 mm/spl times/10 mm tissue section) and overfill the field of view (FOV) of typical microscope objectives require the use of image tiling to cover the entire specimen. This can be time consuming and cause artifacts in the composite image. The MACROscope system (Biomedical Photometrics Inc., Waterloo, ON, Canada) is a confocal device with a 22 mm/spl times/70 mm FOV designed for imaging large tissue sections in a single frame. The prototype demonstrated here can obtain images in reflected, transmitted, fluorescence, phase contrast, and hyperspectral modes. The new spectral imaging mode is characterized with a series of test targets, and sampled spectra are compared to a commercial spectrometer. Fluorescence images of human SiHa tumor xenografts stained with CD31-Cy3, showing blood vessel location, and EF5-Cy5, showing areas of tissue hypoxia, were collected. Differential phase contrast images of the same section, as well as human epithelial cells, were recorded to assess the phase contrast mode. Additionally, fluorescence images of Cytokeratin-Cy3 stained squamous cell carcinoma tissue sections were captured. Finally, red, green, blue transmitted light images of human tongue were obtained. This new device avoids the need for image tiling and provides simultaneous imaging of multiple fluorescently labeled tissue-specific markers in large biological samples. This enables time- and cost-efficient imaging of (immuno)histopathological samples. This device may also serve in the imaging of high-throughput DNA and tissue arrays.