Intravital microscopy of biosensor activities and intrinsic metabolic states

Winfree, Seth; Hato, Takashi; Day, Richard N.

doi:10.1016/j.ymeth.2017.04.017

Cited by 10 publications

(3 citation statements)

References 41 publications

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“…IVM is a valuable tool to evaluate a number of different physiological functions (e.g., microvascular flow, mitochondrial function, transport) using fluorescent probes introduced either intravenously or intraperitoneally 8,29 . While studies examining transplanted islet function have been informative 22 , examples of IVM for the study of endogenous β-cell function and signaling in vivo are sparse 4 , likely due to the historical need for transgenic animals expressing fluorescent biosensors.…”

Section: Discussionmentioning

confidence: 99%

A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo

Reissaus

Piñeros

Twigg

et al. 2019

Sci Rep

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The pancreatic islet is a complex micro-organ containing numerous cell types, including endocrine, immune, and endothelial cells. The communication of these systems is lost upon isolation of the islets, and therefore the pathogenesis of diabetes can only be fully understood by studying this organized, multicellular environment in vivo . We have developed several adaptable tools to create a versatile platform to interrogate β-cell function in vivo . Specifically, we developed β-cell-selective virally-encoded fluorescent protein biosensors that can be rapidly and easily introduced into any mouse. We then coupled the use of these biosensors with intravital microscopy, a powerful tool that can be used to collect cellular and subcellular data from living tissues. Together, these approaches allowed the observation of in vivo β-cell-specific ROS dynamics using the Grx1-roGFP2 biosensor and calcium signaling using the GcAMP6s biosensor. Next, we utilized abdominal imaging windows (AIW) to extend our in vivo observations beyond single-point terminal measurements to collect longitudinal physiological and biosensor data through repeated imaging of the same mice over time. This platform represents a significant advancement in our ability to study β-cell structure and signaling in vivo , and its portability for use in virtually any mouse model will enable meaningful studies of β-cell physiology in the endogenous islet niche.

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Section: Discussionmentioning

confidence: 99%

A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo

Reissaus

Piñeros

Twigg

et al. 2019

Sci Rep

Self Cite

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“…One limitation of IVM is overlap or bleed-through between related fluorophores and when using particular laser light sources and microscope filter sets while labeling and imaging more than one component of the TME. To overcome this limitation, fluorescence-lifetime imaging microscopy can be used in which fluorophores with different fluorescence lifetimes are used [ 294 ] and this can be combined with standard wavelength filtering to allow investigation of TME dynamics [ 295 , 296 ] .…”

Section: Imaging the Tumor Microenvironmentmentioning

confidence: 99%

Imaging methods to evaluate tumor microenvironment factors affecting nanoparticle drug delivery and antitumor response

Moody¹,

Dayton²,

Zamboni³

2021

CDR

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Standard small molecule and nanoparticulate chemotherapies are used for cancer treatment; however, their effectiveness remains highly variable. One reason for this variable response is hypothesized to be due to nonspecific drug distribution and heterogeneity of the tumor microenvironment, which affect tumor delivery of the agents. Nanoparticle drugs have many theoretical advantages, but due to variability in tumor microenvironment (TME) factors, the overall drug delivery to tumors and associated antitumor response are low. The nanotechnology field would greatly benefit from a thorough analysis of the TME factors that create these physiological barriers to tumor delivery and treatment in preclinical models and in patients. Thus, there is a need to develop methods that can be used to reveal the content of the TME, determine how these TME factors affect drug delivery, and modulate TME factors to increase the tumor delivery and efficacy of nanoparticles. In this review, we will discuss TME factors involved in drug delivery, and how biomedical imaging tools can be used to evaluate tumor barriers and predict drug delivery to tumors and antitumor response.

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“…If lifetimes are distinct, this information could be used to separate spectrally identical species, further increasing the number of molecular targets that could be imaged[34]. The use of multiphoton fluorescence lifetime imaging microscopy to image intrinsic fluorophores and metabolic state is also an interesting prospect[35–37]. For instance, imaging intrinsic fluorophores with fluorescence lifetime imaging microscopy could provide a method for rapid assessment of clinical samples[38].…”

Section: Introductionmentioning

confidence: 99%

Large-scale 3-dimensional quantitative imaging of tissues: state-of-the-art and translational implications

Winfree

Ferkowicz

Dagher

et al. 2017

Translational Research

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Recent developments in automated optical sectioning microscope systems have enabled researchers to conduct high resolution, three-dimensional (3D) microscopy at the scale of millimeters in various types of tissues. This powerful technology allows the exploration of tissues at an unprecedented level of detail, while preserving the spatial context. By doing so, such technology will also enable researchers to explore cellular and molecular signatures within tissue and correlate with disease course. This will allow an improved understanding of pathophysiology and facilitate a precision medicine approach to assess the response to treatment. The ability to perform large-scale imaging in 3D cannot be realized without the widespread availability of accessible quantitative analysis. In this review, we will outline recent advances in large-scale 3D imaging, and discuss the available methodologies to perform meaningful analysis and potential applications in translational research.

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Intravital microscopy of biosensor activities and intrinsic metabolic states

Cited by 10 publications

References 41 publications

A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo

A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo

Imaging methods to evaluate tumor microenvironment factors affecting nanoparticle drug delivery and antitumor response

Large-scale 3-dimensional quantitative imaging of tissues: state-of-the-art and translational implications

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