scholarly journals Multi-Modal Multi-Spectral Intravital Microscopic Imaging of Signaling Dynamics in Real-Time during Tumor–Immune Interactions

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 499
Author(s):  
Tracy W. Liu ◽  
Seth T. Gammon ◽  
David Piwnica-Worms
Keyword(s):  
Molecular Imaging ◽  
Single Cell ◽  
Real Time ◽  
Bioluminescence Imaging ◽  
Emission Spectra ◽  
Ccd Camera ◽  
Microscopic Imaging ◽  
Signaling Dynamics ◽  
Window Chamber

Intravital microscopic imaging (IVM) allows for the study of interactions between immune cells and tumor cells in a dynamic, physiologically relevant system in vivo. Current IVM strategies primarily use fluorescence imaging; however, with the advances in bioluminescence imaging and the development of new bioluminescent reporters with expanded emission spectra, the applications for bioluminescence are extending to single cell imaging. Herein, we describe a molecular imaging window chamber platform that uniquely combines both bioluminescent and fluorescent genetically encoded reporters, as well as exogenous reporters, providing a powerful multi-plex strategy to study molecular and cellular processes in real-time in intact living systems at single cell resolution all in one system. We demonstrate that our molecular imaging window chamber platform is capable of imaging signaling dynamics in real-time at cellular resolution during tumor progression. Importantly, we expand the utility of IVM by modifying an off-the-shelf commercial system with the addition of bioluminescence imaging achieved by the addition of a CCD camera and demonstrate high quality imaging within the reaches of any biology laboratory.

scholarly journals Multi-Modal Multi-Spectral Intravital Macroscopic Imaging of Signaling Dynamics in Real Time during Tumor–Immune Interactions

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 489 ◽  
Author(s):  
Tracy W. Liu ◽  
Seth T. Gammon ◽  
David Fuentes ◽  
David Piwnica-Worms
Keyword(s):  
Molecular Imaging ◽  
Real Time ◽  
Bioluminescence Imaging ◽  
Immune Cell ◽  
Imaging Techniques ◽  
Spectral Unmixing ◽  
Fluorescent Reporters ◽  
Signaling Dynamics ◽  
Cell Enzyme

A major obstacle in studying the interplay between cancer cells and the immune system has been the examination of proposed biological pathways and cell interactions in a dynamic, physiologically relevant system in vivo. Intravital imaging strategies are one of the few molecular imaging techniques that can follow biological processes at cellular resolution over long periods of time in the same individual. Bioluminescence imaging has become a standard preclinical in vivo optical imaging technique with ever-expanding versatility as a result of the development of new emission bioluminescent reporters, advances in genomic techniques, and technical improvements in bioluminescence imaging and processing methods. Herein, we describe an advance of technology with a molecular imaging window chamber platform that combines bioluminescent and fluorescent reporters with intravital macro-imaging techniques and bioluminescence spectral unmixing in real time applied to heterogeneous living systems in vivo for evaluating tumor signaling dynamics and immune cell enzyme activities concurrently.

scholarly journals Proof of concept of a multimodal intravital molecular imaging system for tumour transpathology investigation

2021 ◽  
Author(s):  
Zhen Liu ◽  
Tao Cheng ◽  
Stephan Düwel ◽  
Ziying Jian ◽  
Geoffrey J. Topping ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Imaging System ◽  
Ex Vivo ◽  
Fiducial Markers ◽  
Proof Of Concept ◽  
Registration Accuracy ◽  
Microscopic Imaging ◽  
Window Chamber

Abstract Background Transpathology highlights the interpretation of the underlying physiology behind molecular imaging. However, it remains challenging due to the discrepancies between in vivo and in vitro measurements and difficulties of precise co-registration between trans-scaled images. This study aims to develop a multimodal intravital molecular imaging (MIMI) system as a tool for in vivo tumour transpathology investigation. Methods The proposed MIMI system integrates high-resolution positron imaging, magnetic resonance imaging (MRI) and microscopic imaging on a dorsal skin window chamber on an athymic nude rat. The window chamber frame was designed to be compatible with multimodal imaging and its fiducial markers were customized for precise physical alignment among modalities. The co-registration accuracy was evaluated based on phantoms with thin catheters. For proof of concept, tumour models of the human colorectal adenocarcinoma cell line HT-29 were imaged. The tissue within the window chamber was sectioned, fixed and haematoxylin–eosin (HE) stained for comparison with multimodal in vivo imaging. Results The final MIMI system had a maximum field of view (FOV) of 18 mm × 18 mm. Using the fiducial markers and the tubing phantom, the co-registration errors are 0.18 ± 0.27 mm between MRI and positron imaging, 0.19 ± 0.22 mm between positron imaging and microscopic imaging and 0.15 ± 0.27 mm between MRI and microscopic imaging. A pilot test demonstrated that the MIMI system provides an integrative visualization of the tumour anatomy, vasculatures and metabolism of the in vivo tumour microenvironment, which was consistent with ex vivo pathology. Conclusions The established multimodal intravital imaging system provided a co-registered in vivo platform for trans-scale and transparent investigation of the underlying pathology behind imaging, which has the potential to enhance the translation of molecular imaging.

scholarly journals Proof of Concept of a Multimodal Intravital Molecular Imaging System for Tumour Transpathology Investigation

2021 ◽  
Author(s):  
Zhen Liu ◽  
Tao Cheng ◽  
Stephan Düwel ◽  
Ziying Jian ◽  
Geoffrey J. Topping ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Imaging System ◽  
Fiducial Markers ◽  
Proof Of Concept ◽  
Registration Accuracy ◽  
Microscopic Imaging ◽  
Registration Errors ◽  
Window Chamber

Abstract Background: Transpathology highlights the interpretation of underlying physiology behind molecular imaging. However, it remains challenging due to the discrepancies between in vivo and in vitro measurements and difficulties of precise co-registration between multi-scaled images. This study aims to develop a multimodal intravital molecular imaging (MIMI) system as a tool for in vivo tumour transpathology investigation.Methods: The proposed MIMI system integrates high-resolution positron imaging, magnetic resonance imaging (MRI) and microscopic imaging on a dorsal skin window chamber on a nude rat. The window chamber frame was designed to be compatible with multimodal imaging and its fiducial markers were customized for precise physical alignment among modalities. The co-registration accuracy was evaluated based on phantoms with thin catheters. For proof of concept, the human colorectal adenocarcinoma cell line HT-29 was imaged. The tissue within the window chamber was sectioned, fixed, hematoxylin-eosin (HE) stained for comparison with multimodal in vivo imaging.Results: With the physical alignment via fiducial markers, the co-registration errors of the MIMI system were 0.18 ± 0.27 mm between MRI and positron imaging, 0.19 ± 0.22 mm between positron imaging and microscopic imaging, and 0.15 ± 0.27 mm between MRI and microscopic imaging. A pilot test demonstrated that the MIMI system provides an integrative visualization of the tumour anatomy, perfusion and metabolism of the in vivo tumour microenvironment.Conclusions: The established MIMI system provided a co-registered in vivo platform for trans-scale and transparent investigation of the underlying pathology behind imaging, which has the potential to enhance the translation of molecular imaging.

Real-time in vivo bioluminescence imaging of drug-induced oxidative stress

2016 ◽  
Vol 258 ◽  
pp. S234
Author(s):  
S. Seyed Forootan ◽  
F. Mutter ◽  
J. Clarke ◽  
A. Kipar ◽  
K. Park ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Real Time ◽  
Bioluminescence Imaging ◽  
Drug Induced ◽  
In Vivo Bioluminescence Imaging

scholarly journals Gold Nanoparticle-Based Fluorescent Theranostics for Real-Time Image-Guided Assessment of DNA Damage and Repair

2019 ◽  
Vol 20 (3) ◽  
pp. 471 ◽  
Author(s):  
Shriya S. Srinivasan ◽  
Rajesh Seenivasan ◽  
Allison Condie ◽  
Stanton L. Gerson ◽  
Yanming Wang ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Real Time ◽  
Targeted Delivery ◽  
Specific Binding ◽  
Imaging Techniques ◽  
Cancer Cell Line ◽  
Molecular Probe ◽  
Biodistribution Studies

Chemotherapeutic dosing, is largely based on the tolerance levels of toxicity today. Molecular imaging strategies can be leveraged to quantify DNA cytotoxicity and thereby serve as a theranostic tool to improve the efficacy of treatments. Methoxyamine-modified cyanine-7 (Cy7MX) is a molecular probe which binds to apurinic/apyrimidinic (AP)-sites, inhibiting DNA-repair mechanisms implicated by cytotoxic chemotherapies. Herein, we loaded (Cy7MX) onto polyethylene glycol-coated gold nanoparticles (AuNP) to selectively and stably deliver the molecular probe intravenously to tumors. We optimized the properties of Cy7MX-loaded AuNPs using optical spectroscopy and tested the delivery mechanism and binding affinity using the DLD1 colon cancer cell line in vitro. A 10:1 ratio of Cy7MX-AuNPs demonstrated a strong AP site-specific binding and the cumulative release profile demonstrated 97% release within 12 min from a polar to a nonpolar environment. We further demonstrated targeted delivery using imaging and biodistribution studies in vivo in an xenografted mouse model. This work lays a foundation for the development of real-time molecular imaging techniques that are poised to yield quantitative measures of the efficacy and temporal profile of cytotoxic chemotherapies.

Real-time in vivo bioluminescence imaging of lentiviral vector–mediated gene transfer in mouse testis

2010 ◽  
Vol 73 (1) ◽  
pp. 129-138 ◽  
Author(s):  
T.S. Kim ◽  
H.S. Choi ◽  
B.Y. Ryu ◽  
G.T. Gang ◽  
S.U. Kim ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Real Time ◽  
Bioluminescence Imaging ◽  
Lentiviral Vector ◽  
Mouse Testis ◽  
In Vivo Bioluminescence Imaging

Real-time in-vivo microscopic imaging of the cervix using confocal laser endomicroscopy

2016 ◽  
Vol 25 (4) ◽  
pp. 335-343 ◽  
Author(s):  
Michel Degueldre ◽  
Jean Vandromme ◽  
Alexander de Wind ◽  
Francesco Feoli
Keyword(s):  
Real Time ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Microscopic Imaging

scholarly journals Molecular Imaging of Inducible VEGF Expression and Tumor Progression in a Breast Cancer Model

2017 ◽  
Vol 42 (1) ◽  
pp. 407-415 ◽  
Author(s):  
Lu Liang ◽  
Zhiwei Yue ◽  
Wei Du ◽  
Yang Li ◽  
Hongyan Tao ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Molecular Imaging ◽  
Cell Survival ◽  
Real Time ◽  
Tumor Angiogenesis ◽  
Bioluminescence Imaging ◽  
Expression System ◽  
Cancer Model ◽  
Vegf Expression ◽  
Breast Cancer Model

Background: Tumor derived vascular endothelial growth factor (VEGF) can stimulate proliferation and migration of endothelial cells and recruit endothelial progenitor cells into tumors for vascular formation via a paracrine manner. Now increasing evidence suggests that VEGF also serves as an autocrine factor promoting cell survival and tumor angiogenesis. Real time visualization of VEGF activity in the early stages of tumor formation using molecular imaging will provide unprecedented insight into the biological processes of cancer. Methods: The mouse breast cancer cell line 4T1 was transfected with an inducible, bidirectional tetracycline (Bi-Tet) promoter driving VEGF and renilla luciferase (Rluc). This was used to quantitatively image conditional switching of VEGF by bioluminescence imaging (BLI) under the control of systemic administration of doxycycline. Simultaneously, 4T1 cells were labelled with the double fusion reporter gene (Fluc-eGFP) to establish a breast cancer model. Results: We found that inducible VEGF could promote proliferation and attenuate apoptosis due to oxidative stress in an autocrine manner in vitro. In vivo studies revealed that induction of VEGF expression during early tumor development not only dramatically enhanced tumor growth but also increased tumor angiogenesis as visualized by BLI. Finally, immunohistochemistry staining confirmed that inducing VEGF expression promoted cell survival and tumor neovascularization. Conclusion: Together the inducible bidirectional tetracycline (Bi-Tet) co-expression system combined with the dual bioluminescence imaging (BLI) system provides a platform to investigate a target gene’s role in the pathologic process of cancer and facilitates noninvasive monitoring of biological responses in real time.

MP46-04 REAL TIME IN VIVO MOLECULAR IMAGING OF NF-κB IN PROSTATE CANCER: ROLE AS PROGNOSTIC BIOMARKER AND THERAPEUTIC TARGET

2015 ◽  
Vol 193 (4S) ◽  
Author(s):  
Eugene Vykhovanets ◽  
Sanjeev Shukla ◽  
Eswar Shankar ◽  
Olena Vykhovanets ◽  
Gregory MacLennan ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Molecular Imaging ◽  
Real Time ◽  
Therapeutic Target ◽  
Prognostic Biomarker
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