Depth-Resolved Low-Coherence Phase Microscopy for Quantitative 3D Imaging of Stained and Unstained Tissues
This approach combines a single-layer dielectric coating on glass slides with refractive-index-matched mounting media to suppress interface reflections while a white-light source is collimated and spectrally tuned (480–700 nm, 1–3 nm resolution) via an acousto-optic tunable filter. A flipping mirror directs illumination into a reflection-mode module—where backscattered tissue waves interfere with reference waves across ~230 wavelengths using a right-angle prism to prevent double transmission—or into a trans-illumination module employing a transmission grating to separate DC and first-order beams for quantitative phase imaging. Integrated bright-field imaging of H&E-stained tissue, a motorized stage for pre- and post-stain imaging of identical regions, derivative- and polynomial-based autofocusing for chromatic aberration correction, affine co-registration of stained and unstained images, and a semi-automatic nuclear segmentation GUI complete the end-to-end workflow. Inverse Fourier transformation of the spectral interferograms produces depth-resolved optical path difference maps that reveal 3D nanoscale architecture in unlabeled tissue.
Description
This technology is differentiated by its seamless integration of reflection and transmission phase contrasts with bright-field histology, enabling label-free, high-throughput 3D analysis in a single platform. Its spectral tuning and autofocusing strategies deliver precise chromatic correction and submicron axial resolution across hundreds of wavelengths, while the motorized stage and affine registration ensure reproducible imaging before and after staining. The automated segmentation GUI accelerates nuclear identification and pathological correlation, setting a new standard for quantitative, multi-modal tissue characterization.Applications
- Label-free tissue pathology imaging- High-throughput histology slide scanning
- 3D cell morphology analysis
- Drug development screening assays
- Surgical margin optical guidance
Advantages
- Label-free, depth-resolved quantitative phase imaging of unstained tissue with nanoscale sensitivity- Integrated multicontrast imaging (reflection-mode spectral interferometry, transmission phase, and bright-field H&E) for comprehensive analysis
- High-throughput acquisition of spectrally tunable white-light data (230 wavelengths in ~20 s) at 1–3 nm resolution
- Minimized interface reflections and chromatic aberrations via dielectric-coated slides, refractive-index-matched media, and autofocus correction
- Automated affine co-registration of unstained and stained images to enable accurate pre- and post-stain comparison
- Semi-automatic nuclear segmentation with GUI support for efficient pathology correlation
IP Status
https://patents.google.com/patent/US9983399B2Related Publications
Uttam, S., Pham, H. V., LaFace, J., Leibowitz, B., Yu, J., Brand, R. E., Hartman, D. J., & Liu, Y. (2015). Early Prediction of Cancer Progression by Depth-Resolved Nanoscale Mapping of Nuclear Architecture from Unstained Tissue Specimens. Cancer Research, 75(22), 4718–4727. https://doi.org/10.1158/0008-5472.can-15-1274
Thota, P. N., Nasibli, J., Kumar, P., Sanaka, M. R., Chak, A., Zhang, X., Liu, X., Uttam, S., & Liu, Y. (2022). Prediction of neoplastic progression in Barrett’s esophagus using nanoscale nuclear architecture mapping: a pilot study. Gastrointestinal Endoscopy, 95(6), 1239–1246. https://doi.org/10.1016/j.gie.2022.01.007
Uttam, S., Hashash, J. G., LaFace, J., Binion, D., Regueiro, M., Hartman, D. J., Brand, R. E., & Liu, Y. (2019). Three-Dimensional Nanoscale Nuclear Architecture Mapping of Rectal Biopsies Detects Colorectal Neoplasia in Patients with Inflammatory Bowel Disease. Cancer Prevention Research, 12(8), 527–538. https://doi.org/10.1158/1940-6207.capr-19-0024