Source 1review2015Frontiers in Microbiology
Toolkit/single-molecule fluorescence
single-molecule fluorescence
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Single-molecule fluorescence provides high resolution spatial distributions of ribosomes and RNA polymerase (RNAP) in live, rapidly growing Escherichia coli.
Usefulness & Problems
Why this is useful
Single-molecule fluorescence is described as providing high-resolution spatial distributions of ribosomes and RNAP in live, rapidly growing E. coli. In this review it underpins conclusions about segregation between ribosomes and nucleoids.; high-resolution spatial mapping of intracellular components in live E. coli; measuring spatial distributions of ribosomes and RNA polymerase
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Single-molecule fluorescence is described as providing high-resolution spatial distributions of ribosomes and RNAP in live, rapidly growing E. coli. In this review it underpins conclusions about segregation between ribosomes and nucleoids.
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high-resolution spatial mapping of intracellular components in live E. coli
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measuring spatial distributions of ribosomes and RNA polymerase
Problem solved
It addresses the need to resolve intracellular spatial organization more clearly than previous widefield fluorescence studies. This is useful for distinguishing ribosome-rich and nucleoid-rich regions.; improves spatial resolution over previous widefield fluorescence studies for intracellular organization measurements
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It addresses the need to resolve intracellular spatial organization more clearly than previous widefield fluorescence studies. This is useful for distinguishing ribosome-rich and nucleoid-rich regions.
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improves spatial resolution over previous widefield fluorescence studies for intracellular organization measurements
Problem links
improves spatial resolution over previous widefield fluorescence studies for intracellular organization measurements
LiteratureIt addresses the need to resolve intracellular spatial organization more clearly than previous widefield fluorescence studies. This is useful for distinguishing ribosome-rich and nucleoid-rich regions.
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It addresses the need to resolve intracellular spatial organization more clearly than previous widefield fluorescence studies. This is useful for distinguishing ribosome-rich and nucleoid-rich regions.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The method requires live-cell fluorescence imaging and observable molecular targets in E. coli. The abstract does not specify the exact labels or microscope configuration.; requires live-cell fluorescence imaging capability; requires fluorescently observable targets such as ribosomes or RNAP
The abstract does not indicate that the method alone establishes causality for the proposed mechanisms. Mechanistic interpretation is supplemented by drug perturbations and modeling.; the abstract does not specify labels, instrumentation, or exact resolution
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Rapid nucleoid contraction after transcription- or translation-halting drug treatment is consistent with the transertion hypothesis.
Time-resolved imaging after rifampicin or chloramphenicol treatment shows nucleoid contraction on the 0-3 minute timescale.
timescale 0-3 min
Single-molecule fluorescence provides high-resolution spatial distributions of ribosomes and RNA polymerase in live, rapidly growing Escherichia coli.
Ribosomes are more strongly segregated from nucleoids in rapidly growing Escherichia coli than previous widefield fluorescence studies suggested.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug single-molecule-fluorescence
Single-molecule fluorescence provides high resolution spatial distributions of ribosomes and RNA polymerase (RNAP) in live, rapidly growing Escherichia coli.
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review summarysupports
Single-molecule fluorescence provides high-resolution spatial distributions of ribosomes and RNA polymerase in live, rapidly growing Escherichia coli.
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spatial organizationsupports
Ribosomes are more strongly segregated from nucleoids in rapidly growing Escherichia coli than previous widefield fluorescence studies suggested.
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Comparisons
Source-stated alternatives
The abstract contrasts these measurements with previous widefield fluorescence studies, which suggested weaker segregation. No other assay alternative is explicitly detailed in the abstract.
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The abstract contrasts these measurements with previous widefield fluorescence studies, which suggested weaker segregation. No other assay alternative is explicitly detailed in the abstract.
Source-backed strengths
provides high resolution spatial distributions in live cells; supports analysis of ribosome and RNAP localization
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provides high resolution spatial distributions in live cells
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supports analysis of ribosome and RNAP localization
Compared with Langendorff perfused heart electrical recordings
single-molecule fluorescence and Langendorff perfused heart electrical recordings address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Compared with native green gel system
single-molecule fluorescence and native green gel system address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
single-molecule fluorescence and sub-picosecond pump-probe analysis of bacteriorhodopsin pigments address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.