Protocol for multiplexed quantification of fluorescent biosensor FRET efficiency using barcoded cells.

Jhen-Wei Wu, Yichu Xu, Yun Chen, Jr-Ming Yang, Chuan-Hsiang Huang

Abstract read

In one paragraph

Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it

0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

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

0 citing papers in PubMed.

No citing paper in PubMed yet.

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

5 authors.

Jhen-Wei WuDepartment of Pathology, Johns Hopkins University, Baltimore, MD 21287, USA; Center for Cell Dynamics, Johns Hopkins University, Baltimore, MD 21218, USA.

Yichu XuDepartment of Pathology, Johns Hopkins University, Baltimore, MD 21287, USA; Center for Cell Dynamics, Johns Hopkins University, Baltimore, MD 21218, USA.

Yun ChenCenter for Cell Dynamics, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.

Jr-Ming YangDepartment of Pathology, Johns Hopkins University, Baltimore, MD 21287, USA; Center for Cell Dynamics, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: jyang38@jhmi.edu.

Chuan-Hsiang HuangDepartment of Pathology, Johns Hopkins University, Baltimore, MD 21287, USA; Center for Cell Dynamics, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Cell Biology, Johns Hopkins University, Baltimore, MD 21205, USA. Electronic address: chuang29@jhmi.edu.

Funding

Tissue/Pathology/Immunology CoreP50CA098252 · JOHNS HOPKINS UNIVERSITY · 2003 to 2025

$10.1M

NCI NIH HHS P50 CA098252NIGMS NIH HHS R01 GM136711NIGMS NIH HHS R35 GM163951NIH HHS S10 OD016374NIH HHS S10 OD023548

6 · The paper itself

Abstract

Genetically encoded fluorescent biosensors frequently use Förster resonance energy transfer (FRET) between donor and acceptor fluorescent proteins (FPs) as readouts for monitoring molecular activities in live cells. Here, we present a protocol for determining FRET efficiency through the simultaneous imaging of multiple FRET biosensors alongside donor and acceptor FPs using spectrally orthogonal fluorescent cell barcodes. We describe steps for culturing and transfecting cells, acquiring and analyzing images, and calculating FRET efficiency. This approach facilitates simultaneous analysis of multiple FRET biosensors. For complete details on the use and execution of this protocol, please refer to Wu et al.

Indexed as

Biotechnology and bioengineeringCell BiologyMicroscopySingle Cell

Identifiers

PMID42531124

DOI10.1016/j.xpro.2026.104756

PMCPMC13449981

What Socratic holds

Textmetadata

LicenceCC BY-NC-ND

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 Socratic graph.