Evidence map›Paper›PMID 31562888›Full record

ArticleJournal of neuroscience methods2019

High-yield, automated intracellular electrophysiology in retinal pigment epithelia.

Colby F Lewallen, Qin Wan, Arvydas Maminishkis, William Stoy, Ilya Kolb, Nathan Hotaling, Kapil Bharti, Craig R Forest

Open access · greenAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of neuroscience methods, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
0.1field-weighted citation impact, top 56% 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

4 citing papers in PubMed, 3 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

8 authors at 2 institutions in 1 country.

Colby F LewallenGeorgia Institute of Technology, G.W. Woodruff School of Mechanical Engineering, Atlanta, GA 30332, USA. Electronic address: clewallen3@gatech.edu.
Qin WanNational Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Arvydas MaminishkisNational Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
William StoyGeorgia Institute of Technology, Wallace H Coulter Department of Biomedical Engineering, Atlanta, GA 30332, USA.
Ilya KolbGeorgia Institute of Technology, Wallace H Coulter Department of Biomedical Engineering, Atlanta, GA 30332, USA; HHMI Janelia Research Campus, Howard Hughes Medical Institute, Ashburn VA 20147, USA.
Nathan HotalingNational Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Kapil BhartiNational Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Craig R ForestGeorgia Institute of Technology, G.W. Woodruff School of Mechanical Engineering, Atlanta, GA 30332, USA.
Georgia Institute of Technology · USNational Institutes of Health · US

Funding

High-throughput robotic analysis of integrated neuronal phenotypesR01EY023173 · NEI · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · PI BOYDEN, EDWARD S., FOREST, CRAIG · 2012 to 2016
$4.3M
Scalable Cell- and Circuit-Targeted ElectrophysiologyR01NS102727 · NINDS · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · PI BOYDEN, EDWARD S., FOREST, CRAIG · 2017 to 2020
$2.2M
In-vivo circuit activity measurement at single cell, sub-threshold resolutionU01MH106027 · NIMH · GEORGIA INSTITUTE OF TECHNOLOGY · PI FOREST, CRAIG, STANLEY, GARRETT B. · 2014 to 2016
$1.5M
NEI NIH HHS R01 EY023173NIMH NIH HHS U01 MH106027NINDS NIH HHS R01 NS102727
6 · The paper itself

Abstract

backgroundRecent advancements with induced pluripotent stem cell-derived (iPSC) retinal pigment epithelium (RPE) have made disease modeling and cell therapy for macular degeneration feasible. However, current techniques for intracellular electrophysiology - used to validate epithelial function - are painstaking and require manual skill; limiting experimental throughput. NEW

methodA five-stage algorithm, leveraging advances in automated patch clamping, systematically derived and optimized, improves yield and reduces skill when compared to conventional, manual techniques.

resultsThe automated algorithm improves yield per attempt from 17% (manually, n = 23) to 22% (automated, n = 120) (chi-squared, p = 0.004). Specifically for RPE, depressing the local cell membrane by 6 μm and electroporating (buzzing) just prior to this depth (5 μm) maximized yield. COMPARISON WITH EXISTING

methodConventionally, intracellular epithelial electrophysiology is performed by manually lowering a pipette with a micromanipulator, blindly, towards a monolayer of cells and spontaneously stopping when the magnitude of the instantaneous measured membrane potential decreased below a predetermined threshold. The new method automatically measures the pipette tip resistance during the descent, detects the cell surface, indents the cell membrane, and briefly buzzes to electroporate the membrane while descending, overall achieving a higher yield than conventional methods.

conclusionsThis paper presents an algorithm for high-yield, automated intracellular electrophysiology in epithelia; optimized for human RPE. Automation reduces required user skill and training while, simultaneously, improving yield. This algorithm could enable large-scale exploration of drug toxicity and physiological function verification for numerous kinds of epithelia.

Indexed as

AlgorithmsElectrophysiologyHumansPatch-Clamp TechniquesRetinal Pigment EpitheliumAutomationEpitheliaIntracellular electrophysiologySharp pipette

Identifiers

PMID31562888
PMCPMC7071944
OpenAlexW2976534733

What OpenQuestion holds

Textmetadata
LicenceTDM
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.