Evidence map›Paper›PMID 33387574›Full record

ArticleJournal of neuroscience methods2021

Determining the optimal expression method for dual-color imaging.

Jacob F Norman, Bahar Rahsepar, Jad Noueihed, John A White

Abstract read
In one paragraph

Article in Journal of neuroscience methods, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing 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

3 citing papers in PubMed.

  1. Reactivation of memory-associated neurons induces downstream suppression of competing neuronal populations.Proceedings of the National Academy of Sciences of the United States of America · 2025
    Article
  2. Article
  3. Article
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

4 authors.

Jacob F NormanDept. of Biomedical Engineering, Center for Systems Neuroscience, Neurophotonics Center, Boston University, Boston, MA, 02215, United States. Electronic address: jfnorman@bu.edu.
Bahar RahseparDept. of Biomedical Engineering, Center for Systems Neuroscience, Neurophotonics Center, Boston University, Boston, MA, 02215, United States.
Jad NoueihedDept. of Biomedical Engineering, Center for Systems Neuroscience, Neurophotonics Center, Boston University, Boston, MA, 02215, United States.
John A WhiteDept. of Biomedical Engineering, Center for Systems Neuroscience, Neurophotonics Center, Boston University, Boston, MA, 02215, United States.

Funding

Synchronization in Noisy, Heterogeneous Excitatory/Inhibitory NetworksR01NS054281 · NINDS · LSU HEALTH SCIENCES CENTER · PI CANAVIER, CARMEN CASTRO · 2005 to 2022
$5.4M
Translational Research in BiomaterialsT32EB006359 · NIBIB · BOSTON UNIVERSITY (CHARLES RIVER CAMPUS) · PI MARK W. GRINSTAFF, Michelle H Teplensky · 2009 to 2026
$4.2M
Pan-neuronal functional imaging and anesthesiaR01GM121457 · NIGMS · BOSTON UNIVERSITY MEDICAL CAMPUS · PI CONNOR, CHRISTOPHER W · 2017 to 2021
$1.7M
Advanced laser scanning confocal microscope for multiple usersS10OD024993 · OD · BOSTON UNIVERSITY (CHARLES RIVER CAMPUS) · PI MERTZ, JEROME · 2018 to 2018
$448k
NIBIB NIH HHS T32 EB006359NIGMS NIH HHS R01 GM121457NIH HHS S10 OD024993NINDS NIH HHS R01 NS054281
6 · The paper itself

Abstract

backgroundFluorescence imaging is a widely used technique that permits for cell-type-specific recording from hundreds of neurons simultaneously. Often, to obtain cell-type-specific recordings from more than one cell type, researchers add an additional fluorescent protein to mark a second neuronal subpopulation. Currently, however, no consensus exists on the best expression method for multiple fluorescent proteins. NEW

methodWe optimized the coexpression of two fluorescent proteins across multiple brain regions and mouse lines.

resultsThe single-virus method, a viral injection in a double transgenic reporter mouse, results in limited fluorescent coexpression. In contrast the double-virus method, injecting a mixture of two viruses in a Cre driver mouse, results in up to 70 % coexpression of the fluorescent markers in vitro. Using the double-virus method allows for population activity recording and neuronal subpopulation determination. COMPARISON WITH EXISTING

methodThe standard for expressing two fluorescent proteins is to use a double transgenic reporter mouse with a single viral injection. Injecting two viruses into a Cre driver mouse resulted in significantly higher coexpression compared to the standard method. This result generalized to multiple brain regions and mouse lines in vitro, as well as in vivo.

conclusionEfficiently coexpressing multiple fluorescent proteins provides population activity while identifying a neuronal subpopulation of interest. The improved coexpression is applicable to a wide breadth of experiments, ranging from engram investigation to voltage imaging.

Indexed as

BrainNeuronsAnimalsFluorescenceGenes, ReporterGreen Fluorescent ProteinsLuminescent ProteinsMiceMice, TransgenicOptical ImagingGreen Fluorescent ProteinsLuminescent ProteinsCalcium imagingCell-type specificityGCaMP

Identifiers

PMID33387574
PMCPMC7987797

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Registered trials

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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.