Source 1review2008Histochemistry and Cell Biology
Toolkit/correlative light and electron microscopy
correlative light and electron microscopy
Also known as: CLEM, correlative imaging
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging.
Usefulness & Problems
Why this is useful
Correlative light and electron microscopy is named as one of the technological developments that enabled these cryo-ET studies. The abstract presents it as part of the supporting workflow rather than a standalone endpoint.; enabling cryo-ET studies on FIB-generated lamellae; Correlative light and electron microscopy links fluorescence microscopy with electron microscopy so that targets of interest can be localized with fluorescence and interpreted in ultrastructural context. The abstract presents it as a bridge enabled by new probe development.; linking fluorescence localization with electron-microscopy ultrastructure; bridging live-cell target tracking and higher-resolution structural imaging
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Correlative light and electron microscopy is named as one of the technological developments that enabled these cryo-ET studies. The abstract presents it as part of the supporting workflow rather than a standalone endpoint.
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enabling cryo-ET studies on FIB-generated lamellae
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Correlative light and electron microscopy links fluorescence microscopy with electron microscopy so that targets of interest can be localized with fluorescence and interpreted in ultrastructural context. The abstract presents it as a bridge enabled by new probe development.
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linking fluorescence localization with electron-microscopy ultrastructure
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bridging live-cell target tracking and higher-resolution structural imaging
Problem solved
It contributes enabling capability within the cryo-ET on lamellae pipeline. The abstract does not further specify the exact subproblem addressed.; supports the overall cryo-ET on lamellae workflow as an enabling technology; It helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.; connects molecularly specific fluorescence readouts to ultrastructural context; helps address the resolution limitation of fluorescence microscopy
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It contributes enabling capability within the cryo-ET on lamellae pipeline. The abstract does not further specify the exact subproblem addressed.
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supports the overall cryo-ET on lamellae workflow as an enabling technology
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It helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.
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connects molecularly specific fluorescence readouts to ultrastructural context
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helps address the resolution limitation of fluorescence microscopy
Problem links
connects molecularly specific fluorescence readouts to ultrastructural context
LiteratureIt helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.
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It helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.
helps address the resolution limitation of fluorescence microscopy
LiteratureIt helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.
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It helps overcome the limited resolution of fluorescence microscopy by connecting fluorescent target information to higher-resolution electron microscopy. This supports interpretation of molecules relative to organelles, membranes, and other cellular structures.
supports the overall cryo-ET on lamellae workflow as an enabling technology
LiteratureIt contributes enabling capability within the cryo-ET on lamellae pipeline. The abstract does not further specify the exact subproblem addressed.
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It contributes enabling capability within the cryo-ET on lamellae pipeline. The abstract does not further specify the exact subproblem addressed.
Published Workflows
Objective: Enable in situ structural studies of the near-native cellular interior and surroundings by combining cryogenic sample preparation, lamella fabrication, tomography, and downstream data processing.
Why it works: The review abstract states that isolated-molecule cryo-EM can lose cellular-context information, whereas cryogenic FIB fabrication creates thin lamellae that permit cryo-ET access to the near-native cellular interior. The workflow therefore combines preservation, thinning, imaging, and analysis to recover in situ structural information.
thin lamella generation from cells and tissuestomographic imaging of cellular interiorspreservation of native molecular interactionsultrarapid sample freezingfocused ion beam lamella fabricationcryo-electron tomographydata processingcorrelative light and electron microscopy
Stages
- 1.Ultrarapid sample freezing(library_build)
The abstract names ultrarapid sample freezing as an enabling technological development for cryo-ET studies on FIB-generated lamellae.
Selection: Prepare samples under cryogenic conditions for downstream lamella fabrication and cryo-ET.
- 2.FIB fabrication of lamellae(library_build)
This stage exists because cryogenic FIB fabrication generates thin lamellae that enable cryo-ET structural studies of the near-native cellular interior and surroundings.
Selection: Generate thin lamellae from cellular samples and tissues for cryo-ET.
- 3.Cryo-electron tomography(functional_characterization)
Tomography is the imaging stage that provides structural information in the cellular environment after lamella preparation.
Selection: Acquire structural information from the near-native cellular interior and surroundings.
- 4.Data processing(secondary_characterization)
The abstract states that cellular cryo-ET benefits from data-processing developments and that more in situ structures are being obtained at increasingly higher resolution.
Selection: Process cryo-ET data to support structure determination and higher-resolution in situ analysis.
- 5.Correlative light and electron microscopy support(secondary_characterization)
The abstract names correlative light and electron microscopy as an enabling technological development for these studies.
Selection: Provide correlative imaging support within the cryo-ET on FIB-lamella workflow.
Objective: Bridge live-cell fluorescence microscopy with electron microscopy so that specific targets can be followed dynamically and then interpreted in higher-resolution ultrastructural context.
Why it works: The review describes complementary strengths: fluorescence microscopy can follow specific targets in living cells, while electron microscopy provides higher resolution and ultrastructural context. New probe development enables these modalities to be connected in a single correlative strategy.
fluorescent labeling of specific targetselectron-microscopy visualization of organelles, membranes, and macromoleculescorrelative imaging enabled by probe compatibility across modalitiesfluorescence microscopyelectron microscopycorrelative imaging
Stages
- 1.Fluorescence microscopy of specific targets in living cells(functional_characterization)
This stage captures target-specific and dynamic information that fluorescence microscopy is well suited to provide.
Selection: Follow specific targets on or in living cells to reveal dynamic localization and/or function.
- 2.Electron microscopy for higher-resolution structural context(confirmatory_validation)
This stage addresses the resolution limitation of fluorescence microscopy and places molecules of interest relative to other cellular structures.
Selection: Use electron microscopy to achieve higher resolution and reveal organelles, membranes, and macromolecules.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
correlative imagingelectron microscopy-based ultrastructural imagingfluorescence-based localizationTechniques
Functional AssayTarget processes
localizationInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: sensor
The abstract supports that it is used alongside cryo-ET and FIB-generated lamella workflows. Specific hardware or registration requirements are not given in the provided text.; used as part of the broader cryo-ET on FIB-lamella workflow; It requires fluorescence microscopy, electron microscopy, and probes or labels that support bridging between the two modalities. The abstract also implies access to specialized microscopy infrastructure.; requires access to both fluorescence microscopy and electron microscopy workflows; requires probes or labeling strategies compatible with correlative imaging
the abstract does not specify exact targeting, registration, or performance advantages; The abstract does not claim that correlative imaging removes all limitations of either modality or specify a universal probe solution. Detailed sample-preparation constraints and performance tradeoffs are not provided here.; depends on suitable probe development to bridge the two modalities
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2review2023Nature Methods
Ranked Claims
Cryogenic electron microscopy of isolated macromolecules can achieve near-atomic resolution but does not provide structural information in the cellular environment where macromolecules perform native functions.
Cryogenic electron microscopy and data processing enable the determination of structures of isolated macromolecules to near-atomic resolution. However, these data do not provide structural information in the cellular environment where macromolecules perform their native functions
Cryogenic focused ion beam fabrication generates thin lamellae from cellular samples and tissues, enabling cryo-electron tomography studies of the near-native cellular interior and surroundings.
Cryogenic focused ion beam (FIB) fabrication generates thin lamellae of cellular samples and tissues, enabling structural studies on the near-native cellular interior and its surroundings by cryogenic electron tomography (cryo-ET).
Cellular cryo-ET benefits from developments in electron microscopes, detectors, and data processing, and more in situ structures are being obtained at increasingly higher resolution.
Cellular cryo-ET benefits from the technological developments in electron microscopes, detectors and data processing, and more in situ structures are being obtained and at increasingly higher resolution.
Ultrarapid sample freezing, FIB fabrication of lamellae, tomography, data processing, and correlative light and electron microscopy are enabling developments for cryo-ET studies on FIB-generated lamellae.
the technological developments in ultrarapid sample freezing, FIB fabrication of lamellae, tomography, data processing and correlative light and electron microscopy that have enabled these studies
Electron microscopy reveals organelles, membranes, and macromolecules and helps localize molecules of interest relative to other cellular structures.
Moreover, electron microscopy reveals organelles, membranes, macromolecules, and thus aids in the understanding of cellular complexity and localization of molecules of interest in relation to other structures.
Electron microscopy is the traditional and most well-known way to achieve higher-resolution imaging than fluorescence microscopy.
The traditional and most well-known way to achieve higher resolution imaging is by electron microscopy.
New probe development is building a bridge between fluorescence microscopy and electron microscopy and is enabling correlative imaging.
With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging.
Approval Evidence
2 sources4 linked approval claimsfirst-pass slug correlative-light-and-electron-microscopy
In this Review, we discuss recent studies employing cryo-ET on FIB-generated lamellae and the technological developments in ultrarapid sample freezing, FIB fabrication of lamellae, tomography, data processing and correlative light and electron microscopy that have enabled these studies.
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With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging.
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workflow enablersupports
Ultrarapid sample freezing, FIB fabrication of lamellae, tomography, data processing, and correlative light and electron microscopy are enabling developments for cryo-ET studies on FIB-generated lamellae.
the technological developments in ultrarapid sample freezing, FIB fabrication of lamellae, tomography, data processing and correlative light and electron microscopy that have enabled these studies
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capabilitysupports
Electron microscopy reveals organelles, membranes, and macromolecules and helps localize molecules of interest relative to other cellular structures.
Moreover, electron microscopy reveals organelles, membranes, macromolecules, and thus aids in the understanding of cellular complexity and localization of molecules of interest in relation to other structures.
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comparative capabilitysupports
Electron microscopy is the traditional and most well-known way to achieve higher-resolution imaging than fluorescence microscopy.
The traditional and most well-known way to achieve higher resolution imaging is by electron microscopy.
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trendsupports
New probe development is building a bridge between fluorescence microscopy and electron microscopy and is enabling correlative imaging.
With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging.
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Comparisons
Source-stated alternatives
No explicit alternative correlative targeting approach is named in the abstract.; The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
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No explicit alternative correlative targeting approach is named in the abstract.
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The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Source-backed strengths
identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review
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identified by the review as an enabling technological development
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combines benefits of fluorescence microscopy and electron microscopy
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provides both scientific and practical benefits according to the review
Compared with electron microscopy
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Shared frame: source-stated alternative in extracted literature
Strengths here: identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review.
Relative tradeoffs: the abstract does not specify exact targeting, registration, or performance advantages; depends on suitable probe development to bridge the two modalities.
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The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Compared with fluorescence microscopy
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Shared frame: source-stated alternative in extracted literature
Strengths here: identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review.
Relative tradeoffs: the abstract does not specify exact targeting, registration, or performance advantages; depends on suitable probe development to bridge the two modalities.
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The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Compared with imaging
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Shared frame: source-stated alternative in extracted literature
Strengths here: identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review.
Relative tradeoffs: the abstract does not specify exact targeting, registration, or performance advantages; depends on suitable probe development to bridge the two modalities.
Source:
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Compared with imaging surveillance
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Shared frame: source-stated alternative in extracted literature
Strengths here: identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review.
Relative tradeoffs: the abstract does not specify exact targeting, registration, or performance advantages; depends on suitable probe development to bridge the two modalities.
Source:
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Compared with microscopy
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Shared frame: source-stated alternative in extracted literature
Strengths here: identified by the review as an enabling technological development; combines benefits of fluorescence microscopy and electron microscopy; provides both scientific and practical benefits according to the review.
Relative tradeoffs: the abstract does not specify exact targeting, registration, or performance advantages; depends on suitable probe development to bridge the two modalities.
Source:
The abstract contrasts fluorescence microscopy with electron microscopy as separate modalities, with electron microscopy described as the traditional route to higher resolution. Correlative imaging is presented as the method that bridges them rather than replacing either one.
Ranked Citations
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