Source 1review2018Nature Reviews Physics
Toolkit/fluorescence microscopy
fluorescence microscopy
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Fluorescence microscopy is an imaging assay method used to detect and localize fluorescent signals in living biological specimens. In the supplied evidence, it is described for larval zebrafish as a means to achieve subcellular fluorescence localization and real-time monitoring of cell identity, fate, physiology, and organ pathophysiology.
Usefulness & Problems
Why this is useful
This method is useful for visualizing fluorescent reporters in vivo with subcellular spatial resolution. The cited review specifically positions larval zebrafish as well suited for fluorescence-based microscopy of organs including the pancreas and islets of Langerhans, enabling real-time observation of biological state and physiology.
Source:
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
Problem solved
Fluorescence microscopy addresses the need to monitor where fluorescent signals occur within living tissues and how those signals change over time. In the provided context, it supports in vivo analysis of organ pathophysiology and fluorescent readouts of cell identity, fate, and physiology in larval zebrafish.
Source:
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
Problem links
The evidence explicitly supports live biological imaging and real-time monitoring, which fits the longitudinal, same-specimen part of the gap. It is not a nanoscale solution by itself, but it is a plausible low-damage baseline or comparator for testing less destructive live imaging workflows.
adds optical imaging context to AFM-based mechanical measurements
LiteratureIt helps provide spatial or molecular context alongside AFM-derived mechanical information.
Source:
It helps provide spatial or molecular context alongside AFM-derived mechanical information.
selective visualization of processes involving labeled molecules
LiteratureIt solves the problem of selectively visualizing chosen molecular components within cells or tissues.
Source:
It solves the problem of selectively visualizing chosen molecular components within cells or tissues.
Published Workflows
Objective: Use correlative AFM and optical microscopy to investigate molecular interactions and molecular dynamics with complementary nanoscale physical and optical information.
Why it works: The review abstract states that AFM has important limitations, including non-specificity and low imaging speed, and that combining AFM with complementary optical techniques overcomes these limitations by adding information AFM alone cannot provide.
physical interaction detection by AFMoptical/fluorescence-based complementary readoutatomic force microscopyoptical microscopyfluorescence microscopyconfocal microscopysingle-molecule localization microscopy
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
localizationrecombinationtranslationImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The documented implementation context is in vivo imaging in larval zebrafish, including pancreatic and islet studies. The evidence indicates use with fluorescent probes, but it does not provide construct design details, excitation or emission wavelengths, instrumentation parameters, or sample preparation protocols.
The evidence provided is high level and does not specify microscope modality, fluorophores, quantitative performance, imaging depth, temporal resolution, or sensitivity. It also does not directly validate translation or recombination readouts beyond the broader statement that fluorescence can be modulated and localized.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2021Journal of Visualized Experiments
Source 3review2022FEBS Letters
Source 4review2010Biophysical Reviews
Ranked Claims
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review states that larval zebrafish are well matched to fluorescent probes for real-time monitoring of cell identity, fate, and physiology.
We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
The TAEL/C120 system is used to achieve light-inducible gene expression in zebrafish embryos.
In this protocol, an optogenetic expression system is used to achieve light-inducible gene expression in zebrafish embryos.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue light causes TAEL to dimerize, bind C120, and activate transcription.
When illuminated with blue light, TAEL dimerizes, binds to its cognate regulatory element called C120, and activates transcription.
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
Blue-light illumination induces GFP expression detectable after 30 minutes and reaching more than 130-fold induction after 3 hours in transgenic zebrafish embryos using the TAEL/C120 system.
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
peak GFP induction 130 foldtime to first detection of GFP induction 30 min
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The method is described as a versatile and easy-to-use approach for optogenetic gene expression.
This method is a versatile and easy-to-use approach for optogenetic gene expression.
The review describes AFM as being integrated with complementary methods including optical or fluorescence microscopy, mechanosensitive fluorescent constructs, patch-clamp electrophysiology, and microstructured or fluidic devices.
This review focuses on atomic force microscopy-based mechanobiology and emphasizes AFM modalities for mapping dynamic mechanical properties of biological samples.
Fluorescence microscopy enables specific labeling of biomolecules or supramolecular structures with fluorophores so that images report on processes involving the labeled molecules.
The use of fluorescence microscopy is advantageous because biomolecules or supramolecular structures of interest can be labeled specifically with fluorophores, so the images reveal information on processes involving only the labeled molecules.
Super-resolution fluorescence microscopy techniques have been developed to circumvent the resolution limitation of regular optical microscopy.
In recent years, however, a variety of super-resolution fluorescence microscopy techniques have been developed that circumvent the resolution limitation.
Approval Evidence
4 sources4 linked approval claimsfirst-pass slug fluorescence-microscopy
fast and efficient modulation and localization of fluorescence at a subcellular level, through fluorescence microscopy
Source:
and by fluorescence microscopy
Source:
The supplied review summary explicitly states that AFM is integrated with complementary methods such as optical microscopy, and related item candidates explicitly name fluorescence microscopy.
Source:
The use of fluorescence microscopy is advantageous because biomolecules or supramolecular structures of interest can be labeled specifically with fluorophores, so the images reveal information on processes involving only the labeled molecules.
Source:
review scope summarysupports
Larval zebrafish enable in vivo microscopy for studying organ pathophysiology, including the pancreas and islets of Langerhans.
zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes.
Source:
translational positioningsupports
The review positions living larval zebrafish as a powerful translational research tool and forecasts replacement of many cell line-based studies for understanding organ pathophysiology in whole organisms.
These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line-based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.
Source:
method integrationsupports
The review describes AFM as being integrated with complementary methods including optical or fluorescence microscopy, mechanosensitive fluorescent constructs, patch-clamp electrophysiology, and microstructured or fluidic devices.
Source:
capability summarysupports
Fluorescence microscopy enables specific labeling of biomolecules or supramolecular structures with fluorophores so that images report on processes involving the labeled molecules.
The use of fluorescence microscopy is advantageous because biomolecules or supramolecular structures of interest can be labeled specifically with fluorophores, so the images reveal information on processes involving only the labeled molecules.
Source:
Comparisons
Source-stated alternatives
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.; The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Source:
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Source:
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Source-backed strengths
The supplied evidence supports subcellular localization of fluorescence and real-time monitoring in living larvae. It also indicates compatibility with fluorescent probes in larval zebrafish and applicability to organ-level studies such as the pancreas and islets of Langerhans.
Source:
induction of GFP expression can first be detected after 30 min of illumination and reaches a peak of more than 130-fold induction after 3 h of light treatment
Compared with electrophysiology
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Compared with imaging
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Compared with imaging surveillance
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Compared with microscopy
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.; The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Source:
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Compared with patch-clamp electrophysiology
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Compared with super-resolution fluorescence microscopy
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
The abstract places it within optical microscopy generally and contrasts conventional-resolution imaging with super-resolution fluorescence microscopy techniques.
Compared with super-resolution microscopy
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Shared frame: source-stated alternative in extracted literature
Strengths here: supports multimodal integration with AFM; specific labeling of biomolecules or supramolecular structures; reveals information on processes involving only the labeled molecules.
Relative tradeoffs: the provided source text does not specify exact fluorescence modalities or benchmark advantages; regular optical microscopy remains limited to about half the wavelength of light (~200 nm).
Source:
Other complementary approaches named in the supplied material include patch-clamp electrophysiology, mechanosensitive fluorescent constructs, and super-resolution microscopy.
Ranked Citations
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