Label- and Slide-Free Multispectral Quantitative Epi-Illumination Deep-Ultraviolet Microscopy

Objective and Impact Statement: We present a low-cost, LED-based, epi-illumination deep-UV microscope (epi-DUV) for rapid, label- and slide-free quantitative imaging of fresh bulk tissues, offering a powerful tool for point-of-care and intraoperative histological assessment. Introduction: Label- and slide-free imaging is highly desired in clinical pathology to avoid time-consuming tissue processing while preserving molecular information. Deep-UV microscopy offers high-resolution molecular contrast via intrinsic biomolecular absorption, but prior implementations were limited to thin sections and cell monolayers. Methods: We use an epi-DUV system to capture 255- and 280-nm absorption images, alongside tryptophan autofluorescence, from fresh, unprocessed bulk tissue. A deterministic transformation is applied to the 255-nm absorption images to generate virtual histological staining computationally. Results: This method yields quantitative maps of nucleic acid mass, protein mass, and quantum yield, while the transformation produces accurate H&E-like contrast. The system achieves 0.5-μm lateral resolution and ~6-μm effective slice thickness across a 707 × 707-μm field of view with ~330-ms exposure. This is the first demonstration of quantitative deep-UV molecular imaging of fresh, unlabeled bulk tissue. Conclusion: These results establish a simple, low-cost platform for rapid histological assessment of unprocessed tissue. By providing complementary molecular readouts alongside morphological contrast, this approach overcomes conventional processing limitations for rapid clinical applications.