Label-Free In Vivo Histopathology of Experimental Colitis via 3-Channel Multiphoton Endomicroscopy

Kreiß, Lucas; Thoma, Oana‐Maria; Dilipkumar, Ashwathama; Carlé, Birgitta; Longequeue, Pascal; Kunert, Timo; Räth, Timo; Hildner, Kai; Neufert, Clemens; Friedrich, Oliver; Schürmann, Sebastian; Waldner, Maximilian J.

doi:10.1053/j.gastro.2020.05.081

Cited by 11 publications

(16 citation statements)

References 8 publications

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“…The cecum and colon are the main regions of the conversion of primary BAs into more than 20 secondary BAs through an extensive microbial process ( 37 ). The peristaltic velocity of the GIT is related to the production of 5-hydroxytryptamine by the microbiota in the colon, so there is a bidirectional communication between the host and the microbiota in the colon ( 38 , 39 ). Our results also show that the metabolism of secondary metabolites, energy metabolism, cell growth, and death are the main functions of microbiota in the colon.…”

Section: Discussionmentioning

confidence: 99%

The Lumenal Microbiota Varies Biogeographically in the Gastrointestinal Tract of Rhesus Macaques

et al. 2022

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Section: Discussionmentioning

confidence: 99%

The Lumenal Microbiota Varies Biogeographically in the Gastrointestinal Tract of Rhesus Macaques

et al. 2022

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“…85 An endoscopic application of this system, named multiphoton endomicroscopy (MPEM), has recently been proven to yield label-free in vivo histopathologic results in experimental colitis models. 86…”

Section: Molecular Imagingmentioning

confidence: 99%

Rediscovering histology: what is new in endoscopy for inflammatory bowel disease?

Solitano

D’Amico

Allocca

et al. 2021

Therap Adv Gastroenterol

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The potential of endoscopic evaluation in the management of inflammatory bowel diseases (IBD) has undoubtedly grown over the last few years. When dealing with IBD patients, histological remission (HR) is now considered a desirable target along with symptomatic and endoscopic remission, due to its association with better long-term outcomes. Consequently, the ability of endoscopic techniques to reflect microscopic findings in vivo without having to collect biopsies has become of upmost importance. In this context, a more accurate evaluation of inflammatory disease activity and the detection of dysplasia represent two mainstay targets for IBD endoscopists. New diagnostic technologies have been developed, such as dye-less chromoendoscopy, endomicroscopy, and molecular imaging, but their real incorporation in daily practice is not yet well defined. Although dye-chromoendoscopy is still recommended as the gold standard approach in dysplasia surveillance, recent research questioned the superiority of this technique over new advanced dye-less modalities [narrow band imaging (NBI), Fuji intelligent color enhancement (FICE), i-scan, blue light imaging (BLI) and linked color imaging (LCI)]. The endoscopic armamentarium might also be enriched by new video capsule endoscopy for monitoring disease activity, and high expectations are placed on the application of artificial intelligence (AI) systems to reduce operator-subjectivity and inter-observer variability. The goal of this review is to provide an updated insight on contemporary knowledge regarding new endoscopic techniques and devices, with special focus on their role in the assessment of disease activity and colorectal cancer surveillance.

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“…Optical technologies, such as multiphoton microscopy (MPM), have lately received increasing interest as label-free options to investigate the native tissue morphology of healthy and inflamed colon based on autofluorescence [5,6]. Autofluorescence is a cell property that depends on the presence of endogenous fluorescent molecules, such as NADH and FAD [7].…”

Section: Introductionmentioning

confidence: 99%

“…Using this natural autofluorescence is highly advantageous, as labelfree imaging strategies do not affect the state of individual cells. While MPM allows for the identification of intrastromal cells in colitis models [5], a clear classification of infiltrating immune cell subtypes and functional states is not yet available. Therefore, laying a foundation for understanding how autofluorescence signals differ in individual types of immune cells and their activation status is relevant for possible clinical applications in IBD patients.…”

Section: Introductionmentioning

confidence: 99%

“…MPM is a promising optical imaging technique exploiting the non-linear excitation of fluorescent molecules including NADH and FAD [12]. This allows successful label-free evaluation of 3D tissue morphology, as we have described in previous studies [5]. The specific characteristics of NADH and FAD autofluorescence in single immune cell types and under different functional conditions, however, remain to be described.…”

Section: Introductionmentioning

confidence: 99%

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Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells

Lemire

Thoma

Kreiß

et al. 2022

IJMS

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Immune cell activity is a major factor for disease progression in inflammatory bowel diseases (IBD). Classifying the type and functional state of immune cells is therefore crucial in clinical diagnostics of IBD. Label-free optical technologies exploiting NADH and FAD autofluorescence, such as multiphoton microscopy, have been used to describe tissue morphology in healthy and inflamed colon samples. Nevertheless, a strategy for the identification of single immune cell subtypes within the tissue is yet to be developed. This work aims to initiate an understanding of autofluorescence changes depending on immune cell type and activation state. For this, NADH and FAD autofluorescence signals of different murine immune cell subtypes under native conditions, as well as upon in vitro stimulation and cell death, have been evaluated. Autofluorescence was assessed using flow cytometry and multiphoton microscopy. Our results reveal significantly increased NADH and FAD signals in innate immune cells compared to adaptive immune cells. This allowed identification of relative amounts of neutrophils and CD4+ T cells in mixed cell suspensions, by using NADH signals as a differentiation marker. Furthermore, in vitro stimulation significantly increased NADH and FAD autofluorescence in adaptive immune cells and macrophages. Cell death induced a significant drop in NADH autofluorescence, while FAD signals were hardly affected. Taken together, these results demonstrate the value of autofluorescence as a tool to characterize immune cells in different functional states, paving the way to the label-free clinical classification of IBD in the future.

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Label-Free In Vivo Histopathology of Experimental Colitis via 3-Channel Multiphoton Endomicroscopy

Cited by 11 publications

References 8 publications

The Lumenal Microbiota Varies Biogeographically in the Gastrointestinal Tract of Rhesus Macaques

The Lumenal Microbiota Varies Biogeographically in the Gastrointestinal Tract of Rhesus Macaques

Rediscovering histology: what is new in endoscopy for inflammatory bowel disease?

Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells

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