Source 1primary paper2018The Plant Journal
Toolkit/simultaneous fluorescence intensity analysis software for neighboring cells
simultaneous fluorescence intensity analysis software for neighboring cells
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
To simplify measurements of fluorescence intensity in a complex tissue we developed software that allows simultaneous analyses of fluorescence intensities of several neighbouring cells.
Usefulness & Problems
Why this is useful
The software allows simultaneous analysis of fluorescence intensities from several neighboring cells in complex tissue.; simultaneous fluorescence quantification across several neighboring cells; simplifying analysis in complex tissues
Source:
The software allows simultaneous analysis of fluorescence intensities from several neighboring cells in complex tissue.
Source:
simultaneous fluorescence quantification across several neighboring cells
Source:
simplifying analysis in complex tissues
Problem solved
It simplifies fluorescence intensity measurements in complex tissues where multiple adjacent cells must be analyzed together.; reduces analysis complexity when measuring fluorescence intensities in complex tissue
Source:
It simplifies fluorescence intensity measurements in complex tissues where multiple adjacent cells must be analyzed together.
Source:
reduces analysis complexity when measuring fluorescence intensities in complex tissue
Problem links
reduces analysis complexity when measuring fluorescence intensities in complex tissue
LiteratureIt simplifies fluorescence intensity measurements in complex tissues where multiple adjacent cells must be analyzed together.
Source:
It simplifies fluorescence intensity measurements in complex tissues where multiple adjacent cells must be analyzed together.
Published Workflows
Objective: Quantify passive movement of protein between selected plant cells to investigate symplasmic cell-to-cell connectivity in Arabidopsis root tissue.
Why it works: Selected cells can be optically activated using photoswitchable DRONPA-s, after which movement into neighboring cells can be compared by fluorescence measurements.
plasmodesmata-mediated symplasmic transportreversible photoswitching of DRONPA-s fluorescencetransgenic expressionconfocal microscopyphotoactivation/photoswitchingfluorescence intensity analysis
Steps
- 1.Express DRONPA-s in transgenic Arabidopsisengineered fluorescent reporter system
Provide a photoswitchable fluorescent protein in Arabidopsis root cells for movement measurements.
The reporter must be present in plant cells before selected cells can be optically activated and tracked.
- 2.Activate DRONPA-s fluorescence in selected root meristem cells by confocal illuminationphotoswitchable reporter being activated
Create a localized fluorescent starting population in selected cells for subsequent movement comparison.
Localized activation is needed before neighboring-cell movement can be measured.
- 3.Compare movement of DRONPA-s from activated cells into neighboring cellsreporter system and fluorescence analysis software
Quantify passive intercellular protein movement and compare connectivity between cell types.
Movement can only be assessed after localized activation has established the source cells.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
fluorescence intensity quantificationTechniques
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 abstract implies that fluorescence imaging data from the DRONPA-s assay are required, but does not provide further software or hardware details.; requires fluorescence imaging data from neighboring cells
The abstract does not indicate that the software performs biological interpretation beyond fluorescence intensity analysis.; software is unnamed in the abstract; implementation details and availability are not provided in the abstract
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
In the Arabidopsis root meristem assay, pericycle cells showed the highest efflux capacity with good lateral connectivity.
Our analyses showed that pericycle cells display the highest efflux capacity with a good lateral connectivity.
In the Arabidopsis root meristem assay, root cap cells showed the lowest efflux of DRONPA-s.
In contrast, root cap cells showed the lowest efflux of DRONPA-s.
Quiescent centre plasmodesmata mediated stronger efflux into columella cells than into stele initials.
Plasmodesmata of quiescent centre cells mediated a stronger efflux into columella cells than into stele initials.
The developed software allows simultaneous analysis of fluorescence intensities of several neighboring cells to simplify measurements in complex tissue.
To simplify measurements of fluorescence intensity in a complex tissue we developed software that allows simultaneous analyses of fluorescence intensities of several neighbouring cells.
The DRONPA-s system provides a non-invasive method to quantify passive movement of protein between selected cells, including in deeper tissue layers.
Here we report on an elegant non-invasive method to quantify the passive movement of protein between selected cells even in deeper tissue layers.
The DRONPA-s system is based on a fluorescent protein that can be switched on and off repeatedly by illumination with different light qualities.
The system is based on the fluorescent protein DRONPA-s, which can be switched on and off repeatedly by illumination with different light qualities.
The DRONPA-s system generates reproducible data and is a valuable tool for studying symplasmic connectivity.
Our DRONPA-s system generates reproducible data and is a valuable tool for studying symplasmic connectivity.
Approval Evidence
1 source1 linked approval claimfirst-pass slug simultaneous-fluorescence-intensity-analysis-software-for-neighboring-cells
To simplify measurements of fluorescence intensity in a complex tissue we developed software that allows simultaneous analyses of fluorescence intensities of several neighbouring cells.
Source:
tool capabilitysupports
The developed software allows simultaneous analysis of fluorescence intensities of several neighboring cells to simplify measurements in complex tissue.
To simplify measurements of fluorescence intensity in a complex tissue we developed software that allows simultaneous analyses of fluorescence intensities of several neighbouring cells.
Source:
Comparisons
Source-stated alternatives
No alternative analysis software or manual workflow is explicitly named in the abstract.
Source:
No alternative analysis software or manual workflow is explicitly named in the abstract.
Source-backed strengths
supports simultaneous analysis of several neighboring cells
Source:
supports simultaneous analysis of several neighboring cells
Compared with Langendorff perfused heart electrical recordings
simultaneous fluorescence intensity analysis software for neighboring cells 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
simultaneous fluorescence intensity analysis software for neighboring cells 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.
simultaneous fluorescence intensity analysis software for neighboring cells 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.