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Proceedings Paper

Influence of sample preparation and reliability of automated numerical refocusing in stain-free analysis of dissected tissues with quantitative phase digital holographic microscopy
Author(s): Björn Kemper; Philipp Lenz; Dominik Bettenworth; Philipp Krausewitz; Dirk Domagk; Steffi Ketelhut
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Paper Abstract

Digital holographic microscopy (DHM) has been demonstrated to be a versatile tool for high resolution non-destructive quantitative phase imaging of surfaces and multi-modal minimally-invasive monitoring of living cell cultures in-vitro. DHM provides quantitative monitoring of physiological processes through functional imaging and structural analysis which, for example, gives new insight into signalling of cellular water permeability and cell morphology changes due to toxins and infections. Also the analysis of dissected tissues quantitative DHM phase contrast prospects application fields by stain-free imaging and the quantification of tissue density changes. We show that DHM allows imaging of different tissue layers with high contrast in unstained tissue sections. As the investigation of fixed samples represents a very important application field in pathology, we also analyzed the influence of the sample preparation. The retrieved data demonstrate that the quality of quantitative DHM phase images of dissected tissues depends strongly on the fixing method and common staining agents. As in DHM the reconstruction is performed numerically, multi-focus imaging is achieved from a single digital hologram. Thus, we evaluated the automated refocussing feature of DHM for application on different types of dissected tissues and revealed that on moderately stained samples highly reproducible holographic autofocussing can be achieved. Finally, it is demonstrated that alterations of the spatial refractive index distribution in murine and human tissue samples represent a reliable absolute parameter that is related of different degrees of inflammation in experimental colitis and Crohn’s disease. This paves the way towards the usage of DHM in digital pathology for automated histological examinations and further studies to elucidate the translational potential of quantitative phase microscopy for the clinical management of patients, e.g., with inflammatory bowel disease.

Paper Details

Date Published: 22 June 2015
PDF: 6 pages
Proc. SPIE 9529, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II, 952904 (22 June 2015); doi: 10.1117/12.2184681
Show Author Affiliations
Björn Kemper, Westfälische Wilhelms-Univ. Münster (Germany)
Philipp Lenz, Universitätsklinikum Münster (Germany)
Dominik Bettenworth, Universitätsklinikum Münster (Germany)
Philipp Krausewitz, Universitätsklinikum Münster (Germany)
Dirk Domagk, Universitätsklinikum Münster (Germany)
Steffi Ketelhut, Westfälische Wilhelms-Univ. Münster (Germany)


Published in SPIE Proceedings Vol. 9529:
Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II
Pietro Ferraro; Simonetta Grilli; Monika Ritsch-Marte; David Stifter, Editor(s)

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