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Record W3187950054 · doi:10.1002/elps.202100144

Segmentation‐based analysis of single‐cell immunoblots

2021· article· en· W3187950054 on OpenAlexfundno aff
Anjali Gopal, Amy E. Herr

Bibliographic record

VenueElectrophoresis · 2021
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicCell Image Analysis Techniques
Canadian institutionsnot available
FundersNational Institute of General Medical SciencesNatural Sciences and Engineering Research Council of CanadaNational Institutes of HealthNational Cancer InstituteThomas and Stacey Siebel Foundation
KeywordsSegmentationMixture modelQuantitative proteomicsProteomicsComputer scienceProtein subcellular localization predictionPipeline (software)Computational biologyPattern recognition (psychology)Artificial intelligenceBiologyMolecular biologyBiochemistryGene

Abstract

fetched live from OpenAlex

Abstract From genomics to transcriptomics to proteomics, microfluidic tools underpin recent advances in single‐cell biology. Detection of specific proteoforms—with single‐cell resolution—presents challenges in detection specificity and sensitivity. Miniaturization of protein immunoblots to single‐cell resolution mitigates these challenges. For example, in microfluidic western blotting, protein targets are separated by electrophoresis and subsequently detected using fluorescently labeled antibody probes. To quantify the expression level of each protein target, the fluorescent protein bands are fit to Gaussians; yet, this method is difficult to use with noisy, low‐abundance, or low‐SNR protein bands, and with significant band skew or dispersion. In this study, we investigate segmentation‐based approaches to robustly quantify protein bands from single‐cell protein immunoblots. As compared to a Gaussian fitting pipeline, the segmentation pipeline detects >1.5× more protein bands for downstream quantification as well as more of the low‐abundance protein bands (i.e., with SNR ∼3). Utilizing deep learning‐based segmentation approaches increases the recovery of low‐SNR protein bands by an additional 50%. However, we find that segmentation‐based approaches are less robust at quantifying poorly resolved protein bands (separation resolution, Rs < 0.6). With burgeoning needs for more single‐cell protein analysis tools, we see microfluidic separations as benefitting substantially from segmentation‐based analysis approaches.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.001
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Methods · Consensus signal: Methods
Teacher disagreement score0.002
Threshold uncertainty score0.007

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0010.001
Meta-epidemiology (narrow)0.0010.000
Meta-epidemiology (broad)0.0010.001
Bibliometrics0.0010.001
Science and technology studies0.0000.000
Scholarly communication0.0010.001
Open science0.0010.000
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0020.001

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.005
GPT teacher head0.240
Teacher spread0.234 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreMethods

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations5
Published2021
Admission routes1
Has abstractyes

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