Multimodal Quantitative Phase Imaging with Digital Holographic Microscopy Accurately Assesses Intestinal Inflammation and Epithelial Wound Healing.
The incidence of inflammatory bowel disease, i.e., Crohn's disease and Ulcerative colitis, has significantly increased over the last decade. The etiology of IBD remains unknown and current therapeutic strategies are based on the unspecific suppression of the immune system. The development of treatments that specifically target intestinal inflammation and epithelial wound healing could significantly improve management of IBD, however this requires accurate detection of inflammatory changes. Currently, potential drug candidates are usually evaluated using animal models in vivo or with cell culture based techniques in vitro. Histological examination usually requires the cells or tissues of interest to be stained, which may alter the sample characteristics and furthermore, the interpretation of findings can vary by investigator expertise. Digital holographic microscopy (DHM), based on the detection of optical path length delay, allows stain-free quantitative phase contrast imaging. This allows the results to be directly correlated with absolute biophysical parameters. We demonstrate how measurement of changes in tissue density with DHM, based on refractive index measurement, can quantify inflammatory alterations, without staining, in different layers of colonic tissue specimens from mice and humans with colitis. Additionally, we demonstrate continuous multimodal label-free monitoring of epithelial wound healing in vitro, possible using DHM through the simple automated determination of the wounded area and simultaneous determination of morphological parameters such as dry mass and layer thickness of migrating cells. In conclusion, DHM represents a valuable, novel and quantitative tool for the assessment of intestinal inflammation with absolute values for parameters possible, simplified quantification of epithelial wound healing in vitro and therefore has high potential for translational diagnostic use.
Department of Medicine B, University Hospital Münster; Institute of Palliative Care, University Hospital Münster.
Philipp
P
Lenz
Department of Medicine B, University Hospital Münster.
Markus
M
Brückner
Biomedical Technology Center, University of Münster.
Steffi
S
Ketelhut
Department of Gastroenterology, Klinikum Bielefeld.
Jan
J
Heidemann
Biomedical Technology Center, University of Münster; bkemper@uni-muenster.de.
Björn
B
Kemper
Department of Medicine B, University Hospital Münster.
Dominik
D
Bettenworth
D000818:Animals; D018938:Caco-2 Cells; D002465:Cell Movement / Q000502:physiology; D003093:Colitis, Ulcerative / Q000000981:diagnostic imaging* / Q000473:pathology; D003424:Crohn Disease / Q000000981:diagnostic imaging* / Q000473:pathology; D005260:Female; D006696:Holography / Q000379:methods*; D006801:Humans; D008297:Male; D051379:Mice; D008858:Microscopy, Phase-Contrast / Q000379:methods*; D064847:Multimodal Imaging / Q000379:methods; D014945:Wound Healing / Q000502:physiology*
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