Searching the Resource Information Network

Our searching services are busy right now. Please try again later

  • Register
X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

X

Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.

No
Yes

 PMID:29968564  

Single-color, ratiometric biosensors for detecting signaling activities in live cells.

Brian L Ross | Brian Tenner | Michele L Markwardt | Adam Zviman | Guoli Shi | Jaclyn P Kerr | Nicole E Snell | Jennifer J McFarland | Joseph R Mauban | Christopher W Ward | Megan A Rizzo | Jin Zhang
eLife | 2018

Genetically encoded fluorescent biosensors have revolutionized the study of signal transduction by enabling the real-time tracking of signaling activities in live cells. Investigating the interaction between signaling networks has become increasingly important to understanding complex cellular phenomena, necessitating an update of the biosensor toolkit to allow monitoring and perturbing multiple activities simultaneously in the same cell. We therefore developed a new class of fluorescent biosensors based on homo-FRET, deemed FLuorescence Anisotropy REporters (FLAREs), which combine the multiplexing ability of single-color sensors with a quantitative, ratiometric readout. Using an array of color variants, we were able to demonstrate multiplexed imaging of three activity reporters simultaneously in the same cell. We further demonstrate the compatibility of FLAREs for use with optogenetic tools as well as intravital two-photon imaging.

Pubmed ID: 29968564

Associated grants

  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK073368
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK077140
  • Agency: NIGMS NIH HHS, United States
    Id: T32 GM008181
  • Agency: NIGMS NIH HHS, United States
    Id: R25 GM055036
  • Agency: NCI NIH HHS, United States
    Id: R35 CA197622
  • Agency: NIAMS NIH HHS, United States
    Id: R01 AR071618
  • Agency: NIMH NIH HHS, United States
    Id: R01 MH111527
  • Agency: NIMH NIH HHS, United States
    Id: R01 MH111516
  • Agency: NIH HHS, United States
    Id: R01 MH111516
  • Agency: NIH HHS, United States
    Id: R01 DK073368
  • Agency: NHLBI NIH HHS, United States
    Id: R01 HL122827
  • Agency: NIAMS NIH HHS, United States
    Id: T32 AR007592
  • Agency: NIH HHS, United States
    Id: R01 HL122827
  • Agency: NIH HHS, United States
    Id: R35 CA197622
  • Agency: NIH HHS, United States
    Id: R01 MH111527
  • Agency: NIH HHS, United States
    Id: R01 DK077140

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.

This is a list of tools and resources that we have found mentioned in this publication.


ImageJ (tool)

RRID:SCR_003070

Open source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.

View all literature mentions

GENEWIZ (tool)

RRID:SCR_003177

Commercial organization for research and development genomics services and technical support to researchers.

View all literature mentions

C57BL/6J (tool)

RRID:IMSR_JAX:000664

Mus musculus with name C57BL/6J from IMSR.

View all literature mentions

GraphPad Prism (software resource)

RRID:SCR_002798

Statistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.

View all literature mentions

AxioVision Imaging System (software resource)

RRID:SCR_002677

Digital image processing system where microscope settings and processing steps may be adjusted in single user interface. Can acquire images from variety of cameras. Includes software package for capturing, archiving and preparing images for publication. Allows users to visualize and present images in several dimensions. Functionality of imaging toolbox expands constantly with wide range of different modules that are tailored to specific applications or microscope accessories. This resource is duplicated by SCR_018376

View all literature mentions

REF52 (cell line)

RRID:CVCL_6848

Cell line REF52 is a Spontaneously immortalized cell line with a species of origin Rattus norvegicus (Rat)

View all literature mentions

Fiji (software resource)

RRID:SCR_002285

Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.

View all literature mentions

HEK293T (cell line)

RRID:CVCL_0063

Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)

View all literature mentions

HeLa (cell line)

RRID:CVCL_0030

Cell line HeLa is a Cancer cell line with a species of origin Homo sapiens

View all literature mentions

MATLAB (software resource)

RRID:SCR_001622

Multi paradigm numerical computing environment and fourth generation programming language developed by MathWorks. Allows matrix manipulations, plotting of functions and data, implementation of algorithms, creation of user interfaces, and interfacing with programs written in other languages, including C, C++, Java, Fortran and Python. Used to explore and visualize ideas and collaborate across disciplines including signal and image processing, communications, control systems, and computational finance.

View all literature mentions

MIN6 (cell line)

RRID:CVCL_0431

Cell line MIN6 is a Transformed cell line with a species of origin Mus musculus

View all literature mentions

uManager (software resource)

RRID:SCR_000415

Software package for control of automated microscopes. Cross-platform desktop application, to control motorized microscopes, scientific cameras, stages, illuminators, and other microscope accessories.

View all literature mentions

MetaFluor Fluorescence Ratio Imaging Software (software resource)

RRID:SCR_014294

Software designed for single or multi-wavelength intracellular ion measurements. It provides simultaneous display of raw data, ratio image, graphs of intensities, ratios, ion concentrations, and a non-ratiometric image such as a brightfield or phase-contrast image. Two different ratiometric indicators can be imaged and measured simultaneously to provide greater insight to ion exchange and intracellular function regardless of dye loading concentrations, conditions, or emission intensities.

View all literature mentions