MUSE (Microscopy with UV Surface Excitation) is a simple new approach to microscopy, being a straightforward and inexpensive method that can provide diagnostic-quality images, with enhanced spatial and color information, directly and quickly from fresh or fixed tissue. The process is non-destructive, permitting downstream molecular analyses.
Samples are briefly stained with common fluorescent dyes, followed by 280-nm UV light excitation that generates highly surface-weighted images due to limited penetration depth of light at this wavelength. The method also takes advantage of the "USELESS" phenomenon (UV stain excitation with long emission Stokes shift) for broad-spectrum image generation in the visible range.
MUSE readily provides surface topography information even in single snapshots, and while not fully 3-dimensional, the images are easy to acquire, and easy to interpret, providing more insight into tissue structure.
However, working with samples with intrinsic depth information can pose problems with respect to determining appropriate focal points as well as capturing extended depth-of-field images. We demonstrate an accelerated and efficient variant approach for extending depth of field by employing swept-focus acquisition techniques. We have also developed a novel method for rapid autofocus. Together, these capabilities contribute to MUSE functionality and ease of use.
Farzad Fereidouni, John Tracy, and Richard M. Levenson, "MUSE microscopy for thick tissue imaging with extended depth of field (Conference Presentation)," Proc. SPIE 10489, Optical Biopsy XVI: Toward Real-Time Spectroscopic Imaging and Diagnosis, 104890H (Presented at SPIE BiOS: January 30, 2018; Published: 14 March 2018); https://doi.org/10.1117/12.2291087.5751446480001.
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