scholarly journals Label-free concurrent 5-modal microscopy (Co5M) resolves unknown spatio-temporal processes in wound healing

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Markus Seeger ◽  
Christoph Dehner ◽  
Dominik Jüstel ◽  
Vasilis Ntziachristos
Keyword(s):  
Wound Healing ◽  
Label Free ◽  
Third Harmonic ◽  
Two Photon ◽  
Non Invasive ◽  
Photon Excitation ◽  
Free Mode ◽  
Spatio Temporal ◽  
And Function

AbstractThe non-invasive investigation of multiple biological processes remains a methodological challenge as it requires capturing different contrast mechanisms, usually not available with any single modality. Intravital microscopy has played a key role in dynamically studying biological morphology and function, but it is generally limited to resolving a small number of contrasts, typically generated by the use of transgenic labels, disturbing the biological system. We introduce concurrent 5-modal microscopy (Co5M), illustrating a new concept for label-free in vivo observations by simultaneously capturing optoacoustic, two-photon excitation fluorescence, second and third harmonic generation, and brightfield contrast. We apply Co5M to non-invasively visualize multiple wound healing biomarkers and quantitatively monitor a number of processes and features, including longitudinal changes in wound shape, microvascular and collagen density, vessel size and fractality, and the plasticity of sebaceous glands. Analysis of these parameters offers unique insights into the interplay of wound closure, vasodilation, angiogenesis, skin contracture, and epithelial reformation in space and time, inaccessible by other methods. Co5M challenges the conventional concept of biological observation by yielding multiple simultaneous parameters of pathophysiological processes in a label-free mode.

In vivo label-free two-photon excitation autofluorescence microscopy of microvasculature using a 520 nm femtosecond fiber laser

2020 ◽  
Vol 45 (10) ◽  
pp. 2704
Author(s):  
Ting Wu ◽  
Jiuling Liao ◽  
Jia Yu ◽  
Yufeng Gao ◽  
Hui Li ◽  
...  
Keyword(s):  
Fiber Laser ◽  
Label Free ◽  
Two Photon ◽  
Photon Excitation ◽  
Femtosecond Fiber Laser ◽  
Two Photon Excitation

scholarly journals Label-free imaging of macrophage phenotypes and phagocytic activity in the human dermis in vivo using two-photon excited FLIM

2021 ◽  
Author(s):  
Marius Kröger ◽  
Jörg Scheffel ◽  
Evgeny A. Shirshin ◽  
Johannes Schleusener ◽  
Martina C Meinke ◽  
...  
Keyword(s):  
Fluorescence Lifetime Imaging ◽  
Label Free ◽  
Immune Effector ◽  
Effector Cells ◽  
Two Photon ◽  
Non Invasive ◽  
Human Dermis ◽  
Main Components

Macrophages (MΦs) are important immune effector cells that promote (M1 MΦs) or inhibit (M2 MΦs) inflammation and are involved in numerous physiological and pathogenic immune responses. Their precise role and relevance, however, is not fully understood because of the lack of non-invasive quantification methods. Here, we show that two-photon excited fluorescence lifetime imaging (TPE-FLIM), a label-free non-invasive method, can visualize MΦs in human dermis in vivo. We demonstrate in vitro that human dermal MΦs exhibit specific TPE-FLIM properties that distinguish them from the main components of the extracellular matrix and other dermal cells. We visualized MΦs, their phenotypes and phagocytosis in the skin of healthy individuals in vivo using TPE-FLIM. Additionally, machine learning identified M1 and M2 MΦs with a sensitivity of 0.88±0.04 and 0.82±0.03 and a specificity of 0.89±0.03 and 0.90±0.03, respectively. In clinical research, TPE-FLIM can advance the understanding of the role of MΦs in health and disease.

scholarly journals NMR microsystem for label-free characterization of 3D nanoliter microtissues

2020 ◽  
Author(s):  
Marco Grisi ◽  
Gaurasundar M. Conley ◽  
Kyle J. Rodriguez ◽  
Erika Riva ◽  
Lukas Egli ◽  
...  
Keyword(s):  
Early Stage ◽  
Cmos Technology ◽  
Time Frame ◽  
Ease Of Use ◽  
Chemical Information ◽  
Spectroscopic Techniques ◽  
Label Free ◽  
Alcoholic Fatty Liver ◽  
Non Invasive

AbstractPerforming chemical analysis at the nanoliter (nL) scale is of paramount importance for medicine, drug development, toxicology, and research. Despite the numerous methodologies available, a tool for obtaining chemical information non-invasively is still missing at this scale. Observer effects, sample destruction and complex preparatory procedures remain a necessary compromise1. Among non-invasive spectroscopic techniques, one able to provide holistic and highly resolved chemical information in-vivo is nuclear magnetic resonance (NMR)2,3. For its renowned informative power and ability to foster discoveries and life-saving applications4,5, efficient NMR at microscopic scales is highly sought after6–10, but so far technical limitations could not match the stringent necessities of microbiology, such as biocompatible handling, ease of use, and high throughput. Here we introduce a novel microsystem, which combines CMOS technology with 3D microfabrication, enabling nL NMR as a platform tool for non-invasive spectroscopy of organoids, 3D cell cultures, and early stage embryos. In this study we show its application to microlivers models simulating non-alcoholic fatty liver disease (NAFLD), demonstrating detection of lipid metabolism dynamics in a time frame of 14 days based on 117 measurements of single 3D human liver microtissues.

scholarly journals Enhancing ultrasound texture differences for developing an in vivo 'virtual histology' approach to bovine ovarian imaging

2007 ◽  
Vol 19 (8) ◽  
pp. 910 ◽  
Author(s):  
Mark G. Eramian ◽  
Gregg P. Adams ◽  
Roger A. Pierson
Keyword(s):  
Image Processing ◽  
Ultrasound Image ◽  
Ultrasound Images ◽  
Virtual Histology ◽  
Form And Function ◽  
Non Invasive ◽  
Candidate Image ◽  
And Function ◽  
Processing Techniques

A ‘virtual histology’ can be thought of as the ‘staining’ of a digital ultrasound image via image processing techniques in order to enhance the visualisation of differences in the echotexture of different types of tissues. Several candidate image-processing algorithms for virtual histology using ultrasound images of the bovine ovary were studied. The candidate algorithms were evaluated qualitatively for the ability to enhance the visual differences in intra-ovarian structures and quantitatively, using standard texture description features, for the ability to increase statistical differences in the echotexture of different ovarian tissues. Certain algorithms were found to create textures that were representative of ovarian micro-anatomical structures that one would observe in actual histology. Quantitative analysis using standard texture description features showed that our algorithms increased the statistical differences in the echotexture of stroma regions and corpus luteum regions. This work represents a first step toward both a general algorithm for the virtual histology of ultrasound images and understanding dynamic changes in form and function of the ovary at the microscopic level in a safe, repeatable and non-invasive way.

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