Evidence map›Paper›PMID 28190060›Full record

ArticleJournal of visualized experiments : JoVE2017

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy.

Frederik Görlitz, Douglas J Kelly, Sean C Warren, Dominic Alibhai, Lucien West, Sunil Kumar, Yuriy Alexandrov, Ian Munro, Edwin Garcia, James McGinty and 10 more

Open access · greenAbstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
3.4field-weighted citation impact, top 8% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed, 14 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

20 authors at 4 institutions in 2 countries.

Frederik GörlitzPhotonics Group, Department of Physics, Imperial College London; f.gorlitz14@imperial.ac.uk.
Douglas J KellyPhotonics Group, Department of Physics, Imperial College London.
Sean C WarrenPhotonics Group, Department of Physics, Imperial College London.
Dominic AlibhaiInstitute for Chemical Biology, Department of Chemistry, Imperial College London.
Lucien WestMRC Clinical Sciences Centre, Hammersmith Hospital.
Sunil KumarPhotonics Group, Department of Physics, Imperial College London.
Yuriy AlexandrovPhotonics Group, Department of Physics, Imperial College London.
Ian MunroPhotonics Group, Department of Physics, Imperial College London.
Edwin GarciaPhotonics Group, Department of Physics, Imperial College London.
James McGintyPhotonics Group, Department of Physics, Imperial College London.
Clifford TalbotPhotonics Group, Department of Physics, Imperial College London.
Remigiusz A SerwaChemical Biology Section, Department of Chemistry, Imperial College London.
Emmanuelle ThinonChemical Biology Section, Department of Chemistry, Imperial College London.
Vincenzo da PaolaMRC Clinical Sciences Centre, Hammersmith Hospital.
Edward J MurrayRetroscreen Virology Ltd.
Frank StuhmeierPfizer Global Research and Development, Pfizer Limited, Sandwich, Kent, UK.
Mark A A NeilPhotonics Group, Department of Physics, Imperial College London.
Edward W TateChemical Biology Section, Department of Chemistry, Imperial College London.
Christopher DunsbyPhotonics Group, Department of Physics, Imperial College London; Centre for Histopathology, Imperial College London.
Paul M W FrenchPhotonics Group, Department of Physics, Imperial College London.
Imperial College London · GBPhotonics (United States) · USHammersmith Hospital · GBPfizer (United Kingdom) · GB

Funding

Biotechnology and Biological Sciences Research Council BB/E003621/1Biotechnology and Biological Sciences Research Council BB/H00713X/1Biotechnology and Biological Sciences Research Council BB/M006786/1Biotechnology and Biological Sciences Research Council WT 095931/Z/11/ZCancer Research UK 20183Medical Research Council MC_U120088464Medical Research Council MR/K015834/1Wellcome Trust
6 · The paper itself

Abstract

We present an open source high content analysis instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using Förster resonance energy transfer (FRET) based readouts of fixed or live cells in multiwell plates. This provides a means to screen for cell signaling processes read out using intramolecular FRET biosensors or intermolecular FRET of protein interactions such as oligomerization or heterodimerization, which can be used to identify binding partners. We describe here the functionality of this automated multiwell plate FLIM instrumentation and present exemplar data from our studies of HIV Gag protein oligomerization and a time course of a FRET biosensor in live cells. A detailed description of the practical implementation is then provided with reference to a list of hardware components and a description of the open source data acquisition software written in µManager. The application of FLIMfit, an open source MATLAB-based client for the OMERO platform, to analyze arrays of multiwell plate FLIM data is also presented. The protocols for imaging fixed and live cells are outlined and a demonstration of an automated multiwell plate FLIM experiment using cells expressing fluorescent protein-based FRET constructs is presented. This is complemented by a walk-through of the data analysis for this specific FLIM FRET data set.

Indexed as

AnimalsBiosensing TechniquesChlorocebus aethiopsCOS CellsFluorescence Resonance Energy Transfergag Gene Products, Human Immunodeficiency VirusHumansMicroscopy, FluorescenceOptical ImagingSoftwaregag Gene Products, Human Immunodeficiency Virus

Identifiers

PMID28190060
PMCPMC5352269
OpenAlexW2577225791

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.