Source 1review2023Nature Methods
Toolkit/cryogenic focused ion beam fabrication of lamellae
cryogenic focused ion beam fabrication of lamellae
Also known as: cryogenic FIB fabrication, FIB fabrication of lamellae
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
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).
Usefulness & Problems
No literature-backed usefulness or problem-fit explainer has been materialized for this record yet.
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.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Mechanisms
focused ion beam millingTechniques
Structural CharacterizationTarget processes
No target processes tagged yet.
Input: Light
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
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
Approval Evidence
1 source2 linked approval claimsfirst-pass slug cryogenic-focused-ion-beam-fabrication-of-lamellae
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).
Source:
enabling rolesupports
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).
Source:
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
Source:
Comparisons
No literature-backed comparison notes have been materialized for this record yet.
Ranked Citations
- 1.