Ex vivo and in vivo label-free imaging of lymphatic vessels using OCT lymphangiography (Conference Presentation)

2017 ◽  
Author(s):  
Peijun Gong ◽  
Shaghayegh Es’haghian ◽  
Karol Karnowski ◽  
Suzanne Rea ◽  
Fiona M. Wood ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Label Free

scholarly journals Applications of Vibrational Spectroscopy for Analysis of Connective Tissues

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 922
Author(s):  
William Querido ◽  
Shital Kandel ◽  
Nancy Pleshko
Keyword(s):  
Raman Spectroscopy ◽  
Vibrational Spectroscopy ◽  
Near Infrared ◽  
Ex Vivo ◽  
Biological Tissues ◽  
Label Free ◽  
Connective Tissues ◽  
Bone Properties ◽  
Composition And Structure

Advances in vibrational spectroscopy have propelled new insights into the molecular composition and structure of biological tissues. In this review, we discuss common modalities and techniques of vibrational spectroscopy, and present key examples to illustrate how they have been applied to enrich the assessment of connective tissues. In particular, we focus on applications of Fourier transform infrared (FTIR), near infrared (NIR) and Raman spectroscopy to assess cartilage and bone properties. We present strengths and limitations of each approach and discuss how the combination of spectrometers with microscopes (hyperspectral imaging) and fiber optic probes have greatly advanced their biomedical applications. We show how these modalities may be used to evaluate virtually any type of sample (ex vivo, in situ or in vivo) and how “spectral fingerprints” can be interpreted to quantify outcomes related to tissue composition and quality. We highlight the unparalleled advantage of vibrational spectroscopy as a label-free and often nondestructive approach to assess properties of the extracellular matrix (ECM) associated with normal, developing, aging, pathological and treated tissues. We believe this review will assist readers not only in better understanding applications of FTIR, NIR and Raman spectroscopy, but also in implementing these approaches for their own research projects.

Abstract 27: ApoA-I Improves Lymphatic Function Through a Platelet-Dependent Mechanism in Atherosclerotic Mice

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Andreea Milasan ◽  
François Dallaire ◽  
Gabriel Jean ◽  
Jean-Claude Tardif ◽  
Yahye Merhi ◽  
...  
Keyword(s):  
Lymphatic Vessel ◽  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Lymphatic Transport ◽  
Lv Function ◽  
Experimental Conditions ◽  
Lymphatic Function ◽  
Beneficial Effects ◽  
Dependent Mechanism

Rationale: Lymphatic vessels (LVs) are now recognized as prerequisite players in the modulation of cholesterol removal from the artery wall in experimental conditions of plaque regression, and a particular attention has been brought on the role of the collecting LVs in early atherosclerosis-related lymphatic dysfunction. Whereas recent findings revealed that apoA-I restores the neovascularization capacity of the lymphatic system during tumor necrosis factor-induced inflammation, the effect of apoA-I on collecting LV function during atherosclerosis has not been tested. Objective: In the present study, we address whether and how apoA-I can enhance collecting LV function in atherosclerosis-associated lymphatic dysfunction. Methods and Results: A 6-week systemic treatment with lipid-free apoA-I enhanced lymphatic transport and abrogated collecting lymphatic vessel permeability in atherosclerotic Ldlr –/– mice when compared to control. As injection of apoA-I has been shown to protect wild-type mice against flow restriction-induced thrombosis, and that platelets are identified as key elements in the maintenance of lymphatic vessel integrity via their interaction with lymphatic endothelial cells (LECs), we have tested whether the effects of apoA-I could be mediated through a platelet-dependent mechanism. Our in vivo results show that apoA-I kinetics in lymph reflected that of blood. Ex vivo experiments performed with washed platelets isolated from mouse blood reveal that apoA-I decreased thrombin-induced but not podoplanin-induced platelet aggregation. Whereas this result suggests that apoA-I limits platelet thrombotic potential in blood but not in lymph, we demonstrate that treatment of human LECs with apoA-I increases the adhesion of bridge-like platelets on human LECs. Conclusions: Our results suggest that apoA-I can mediate beneficial effects on lymphatic function by promoting platelet adhesion to the lymphatic endothelium and consequently restore collecting LV integrity. Altogether, we bring forward a new pleiotropic role for apoA-I in lymphatic function and unveil new potential therapeutic targets for the prevention and treatment of atherosclerosis.

scholarly journals In vivo label-free lymphangiography of cutaneous lymphatic vessels in human burn scars using optical coherence tomography

2016 ◽  
Vol 7 (12) ◽  
pp. 4886 ◽  
Author(s):  
Peijun Gong ◽  
Shaghayegh Es’haghian ◽  
Karl-Anton Harms ◽  
Alexandra Murray ◽  
Suzanne Rea ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Lymphatic Vessels ◽  
Optical Coherence ◽  
Label Free

scholarly journals Protocols for Culturing and Imaging a Human Ex Vivo Osteochondral Model for Cartilage Biomanufacturing Applications

Materials ◽  
2019 ◽  
Vol 12 (4) ◽  
pp. 640 ◽  
Author(s):  
Serena Duchi ◽  
Stephanie Doyle ◽  
Timon Eekel ◽  
Cathal D. O’Connell ◽  
Cheryl Augustine ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Cartilage Regeneration ◽  
Light Sheet ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Label Free ◽  
Microscopy Imaging ◽  
Microscopy Technique

Cartilage defects and diseases remain major clinical issues in orthopaedics. Biomanufacturing is now a tangible option for the delivery of bioscaffolds capable of regenerating the deficient cartilage tissue. However, several limitations of in vitro and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. Ex vivo models are of great value for translating in vitro tissue engineered approaches into clinically relevant conditions. Our aim is to obtain a viable human osteochondral (OC) model to test hydrogel-based materials for cartilage repair. Here we describe a detailed step-by-step framework for the generation of human OC plugs, their culture in a perfusion device and the processing procedures for histological and advanced microscopy imaging. Our ex vivo OC model fulfils the following requirements: the model is metabolically stable for a relevant culture period of 4 weeks in a perfusion bioreactor, the processing procedures allowed for the analysis of 3 different tissues or materials (cartilage, bone and hydrogel) without compromising their integrity. We determined a protocol and the settings for a non-linear microscopy technique on label free sections. Furthermore, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical ex vivo tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for in vivo testing.

scholarly journals Label-free 3D imaging of microstructure, blood, and lymphatic vessels within tissue beds in vivo

2012 ◽  
Vol 37 (5) ◽  
pp. 812 ◽  
Author(s):  
Zhongwei Zhi ◽  
Yeongri Jung ◽  
Ruikang K. Wang
Keyword(s):  
3D Imaging ◽  
Lymphatic Vessels ◽  
Label Free

scholarly journals Mesenteric lymph flow in adult and aged rats

2011 ◽  
Vol 301 (5) ◽  
pp. H1828-H1840 ◽  
Author(s):  
Tony J. Akl ◽  
Takashi Nagai ◽  
Gerard L. Coté ◽  
Anatoliy A. Gashev
Keyword(s):  
Shear Stress ◽  
Wall Shear Stress ◽  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Lymph Flow ◽  
Contraction Frequency ◽  
Stress Load ◽  
Wall Shear ◽  
Contractile Characteristics

The objective of study was to evaluate the aging-associated changes, contractile characteristics of mesenteric lymphatic vessels (MLV), and lymph flow in vivo in male 9- and 24-mo-old Fischer-344 rats. Lymphatic diameter, contraction amplitude, contraction frequency, and fractional pump flow, lymph flow velocity, wall shear stress, and minute active wall shear stress load were determined in MLV in vivo before and after Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME) application at 100 μM. The active pumping of the aged rat MLV in vivo was found to be severely depleted, predominantly through the aging-associated decrease in lymphatic contractile frequency. Such changes correlate with enlargement of aged MLV, which experienced much lower minute active shear stress load than adult vessels. At the same time, pumping in aged MLV in vivo may be rapidly increased back to levels of adult vessels predominantly through the increase in contraction frequency induced by nitric oxide (NO) elimination. Findings support the idea that in aged tissues surrounding the aged MLV, the additional source of some yet unlinked lymphatic contraction-stimulatory metabolites is counterbalanced or blocked by NO release. The comparative analysis of the control data obtained from experiments with both adult and aged MLV in vivo and from isolated vessel-based studies clearly demonstrated that ex vivo isolated lymphatic vessels exhibit identical contractile characteristics to lymphatic vessels in vivo.

scholarly journals Brillouin Spectroscopy: From Biomedical Research to New Generation Pathology Diagnosis

2021 ◽  
Vol 22 (15) ◽  
pp. 8055
Author(s):  
Rafael J. Jiménez Rioboó ◽  
Nuria Gontán ◽  
Daniel Sanderson ◽  
Manuel Desco ◽  
Maria Victoria Gómez-Gaviro
Keyword(s):  
Ex Vivo ◽  
Cost Effective ◽  
General View ◽  
Label Free ◽  
Brillouin Spectroscopy ◽  
Innovative Tool ◽  
New Generation ◽  
Further Development ◽  
Biomedical Potential

Brillouin spectroscopy has recently gained considerable interest within the biomedical field as an innovative tool to study mechanical properties in biology. The Brillouin effect is based on the inelastic scattering of photons caused by their interaction with thermodynamically driven acoustic modes or phonons and it is highly dependent on the material’s elasticity. Therefore, Brillouin is a contactless, label-free optic approach to elastic and viscoelastic analysis that has enabled unprecedented analysis of ex vivo and in vivo mechanical behavior of several tissues with a micrometric resolution, paving the way to a promising future in clinical diagnosis. Here, we comprehensively review the different studies of this fast-moving field that have been performed up to date to provide a quick guide of the current literature. In addition, we offer a general view of Brillouin’s biomedical potential to encourage its further development to reach its implementation as a feasible, cost-effective pathology diagnostic tool.

scholarly journals Ex vivo Demonstration of Functional Deficiencies in Popliteal Lymphatic Vessels from TNF-Tg Mice with Inflammatory Arthritis

2020 ◽  
Author(s):  
Joshua P. Scallan ◽  
Echoe M. Bouta ◽  
Homaira Rahimi ◽  
H. Mark Kenney ◽  
Christopher T. Ritchlin ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Repeated Measures ◽  
Inflammatory Arthritis ◽  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Chronic Inflammatory Arthritis ◽  
Wide Range ◽  
Nos Inhibitors ◽  
Draining Lymph Nodes

AbstractBackgroundRheumatoid arthritis (RA) is a progressive immune-mediated inflammatory disease characterized by intermittent episodes of pain and inflammation in affected joints, or flares. Recent studies demonstrated lymphangiogenesis and expansion of draining lymph nodes during chronic inflammatory arthritis, and lymphatic dysfunction associated with collapse of draining lymph nodes in RA patients and TNF-transgenic (TNF-Tg) mice experiencing arthritic flare. As the intrinsic differences between lymphatic vessels afferent to healthy, expanding, and collapsed draining lymph nodes are unknown, we characterized the ex vivo behavior of popliteal lymphatic vessels (PLVs) from WT and TNF-Tg mice. We also interrogated the mechanisms of lymphatic dysfunction through inhibition of nitric oxide synthase (NOS).MethodsPopliteal lymph nodes (PLNs) in TNF-Tg mice were phenotyped as Expanding or Collapsed by in vivo ultrasound and age-matched to WT littermate controls. The PLVs were harvested and cannulated for ex vivo functional analysis over a relatively wide range of hydrostatic pressures (0.5 to 10 cmH2O) to quantify the end diastolic diameter (EDD), tone, amplitude (AMP), ejection fraction (EF), contraction frequency (FREQ) and fractional pump flow (FPF) with or without NOS inhibitors Data was analyzed using repeated measures two-way ANOVA with Bonferroni’s post hoc test.ResultsReal time videos of the cannulated PLVs demonstrated the predicted phenotypes of robust versus weak contractions of the WT versus TNF-Tg PLV, respectively. Quantitative analyses confirmed that TNF-Tg PLVs had significantly decreased AMP, EF and FPF versus WT (p<0.05). EF and FPF were recovered by NOS inhibition, while the reduction in AMP was NOS independent. No differences in EDD, tone, or FREQ were observed between WT and TNF-Tg PLVs, nor between Expanding versus Collapsed PLVs.ConclusionThese findings support the concept that chronic inflammatory arthritis leads to NOS dependent and independent draining lymphatic vessel dysfunction that exacerbates disease, and may trigger arthritic flare due to decreased egress of inflammatory cells and soluble factors from affected joints.

scholarly journals How To Extend The Capabilities Of A Commercial Two-Photon Microscope To Perform Super-Resolution Imaging, Wavelength Mixing And Label-Free Microscopy.

2021 ◽  
Author(s):  
Chiara Peres ◽  
Chiara Nardin ◽  
Guang Yang ◽  
Fabio Mammano
Keyword(s):  
Ex Vivo ◽  
Super Resolution ◽  
Label Free ◽  
Linear Optics ◽  
Non Linear Optics ◽  
Two Photon ◽  
Custom Made ◽  
Non Linear ◽  
Resolution Imaging

Multimodal microscopy combines multiple non-linear techniques that take advantage of different optical processes to generate contrast and increase the amount of information that can be obtained from biological samples. However, the most advanced optical architectures are typically custom-made and require complex alignment procedures, as well as daily maintenance by properly trained personnel for optimal performance. Here, we describe a hybrid system we constructed to overcome these disadvantages by modifying a commercial upright microscope. We show that our multimodal imaging platform can be used to seamlessly perform two-photon STED, wavelength mixing and label-free microscopy in both ex vivo and in vivo samples. The system is highly stable and endowed with remote alignment hardware that ensures simplified operability for non-expert users. This optical architecture is an important step forward towards a wider practical applicability of non-linear optics to bioimaging.

scholarly journals Identification of lymphatic pathway involved in the spread of bladder cancer: Evidence obtained from fluorescence navigation with intraoperatively injected indocyanine green

2013 ◽  
Vol 7 (5-6) ◽  
pp. 322 ◽  
Author(s):  
Shogo Inoue ◽  
Hiroaki Shiina ◽  
Yozo Mitsui ◽  
Hiroaki Yasumoto ◽  
Akio Matsubara ◽  
...  
Keyword(s):  
Indocyanine Green ◽  
Radical Cystectomy ◽  
Fluorescence Intensity ◽  
Ex Vivo ◽  
Lymphatic Vessels ◽  
Rational Approach ◽  
Lymphatic Channel ◽  
Internal Iliac ◽  
Lymphatic Pathway

Introduction: We identify lymphatic vessels draining from the bladderby using fluorescence navigation (FN) system.Methods: In total, 12 candidates for radical cystectomy and pelviclymph node dissection (PLND) were included in this study. Afteran indocyanine green (ICG) solution was injected into the bladderduring radical cystectomy, lymphatic vessels draining from thebladder were analyzed using a FN system. PLND was based onthe lymphatic mapping created from the FN measurements (in vivoprobing) in the external iliac, obturator and internal iliac regions;after PLND, the fluorescence of the removed lymph nodes (LNs)was analyzed on the bench (ex vivo probing).Results: There were no patients with complications associated withthe intravesical ICG injection. A lymphatic pathway along inferiorvesical vessels to internal iliac LNs was clearly illustrated in 7 cases.Under in-vivo probing, the fluorescence intensity of internal iliacnodes was greater than that of external iliac or obturator nodes.Under ex-vivo probing, the fluorescence intensity of internal iliacand obturator nodes was greater than that of external iliac nodes.Conclusions: Using an FN system after injecting ICG during a radicalcystectomy operation is a safe and rational approach to detectingthe lymphatic channel draining from the bladder.

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