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Biomed. Opt."],"published-print":{"date-parts":[[2019,2,8]]},"abstract":"<jats:p>Intraoperative assessment of breast surgical margins will be of value for reducing the rate of re-excision surgeries for lumpectomy patients. While frozen-section histology is used for intraoperative guidance of certain cancers, it provides limited sampling of the margin surface (typically &lt;1\u2009\u2009%\u2009\u2009 of the margin) and is inferior to gold-standard histology, especially for fatty tissues that do not freeze well, such as breast specimens. Microscopy with ultraviolet surface excitation (MUSE) is a nondestructive superficial optical-sectioning technique that has the potential to enable rapid, high-resolution examination of excised margin surfaces. Here, a MUSE system is developed with fully automated sample translation to image fresh tissue surfaces over large areas and at multiple levels of defocus, at a rate of \u223c5\u2009\u2009min\u2009\u2009\/\u2009\u2009cm2. Surface extraction is used to improve the comprehensiveness of surface imaging, and 3-D deconvolution is used to improve resolution and contrast. In addition, an improved fluorescent analog of conventional H&amp;E staining is developed to label fresh tissues within \u223c5\u2009\u2009min for MUSE imaging. 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