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Record W2329142534 · doi:10.1101/pdb.prot080226

Fluorescent Visualization of Germline Apoptosis in Living <i>Caenorhabditis elegans</i>

2014· article· en· W2329142534 on OpenAlexaff
Benjamin Lant, W. Brent Derry

Bibliographic record

VenueCold Spring Harbor Protocols · 2014
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicGenetics, Aging, and Longevity in Model Organisms
Canadian institutionsUniversity of TorontoHospital for Sick Children
Fundersnot available
KeywordsCaenorhabditis elegansApoptosisGreen fluorescent proteinAcridine orangeCell biologyBiologyRNA interferenceFluorescenceFluorescence microscopeProgrammed cell deathHigh-content screeningCellMolecular biologyGeneRNAGenetics

Abstract

fetched live from OpenAlex

Visualization of apoptosis using fluorescent tools is quite straightforward in living Caenorhabditis elegans. Several transgenic lines are available that mark dying cells with fluorescent proteins, making it possible to quantify kinetics at various stages of the apoptotic process. Proteins required for the engulfment of cell corpses are particularly useful for detecting early apoptotic stages using this approach. For example, expression of the engulfment protein CED-1 fused to green fluorescent protein (CED-1::GFP) creates a halo around cells during early apoptosis, before their refractile morphology can be detected by differential interference contrast (DIC) optics. In addition, vital dyes such as acridine orange (AO) and SYTO-12 are selectively retained in apoptotic cells and can be used to visualize apoptosis in the germlines of living animals. It is also possible to use vital dyes in combination with transgenic strains expressing fluorescent markers of cell corpses to examine, in detail, multiple stages of apoptosis in vivo. Because of the high optical contrast of fluorescent reagents, apoptosis can be visualized even at low magnification, facilitating the use of screening platforms to identify apoptosis regulators. This protocol describes multiple uses of fluorescent reagents for visualization of germline apoptosis in living C. elegans, including AO staining, time-course studies using fluorescent proteins, and low-throughput screening of cell death genes using RNA interference (RNAi).

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)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Not applicable · Consensus signal: none
GenreCandidate signal: Methods · Consensus signal: none
Teacher disagreement score0.004
Threshold uncertainty score0.011

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0010.000
Science and technology studies0.0010.000
Scholarly communication0.0000.000
Open science0.0010.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0030.001

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 head0.009
GPT teacher head0.254
Teacher spread0.245 · 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 designNot applicable
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

Citations30
Published2014
Admission routes1
Has abstractyes

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Same venueCold Spring Harbor ProtocolsSame topicGenetics, Aging, and Longevity in Model OrganismsFrench-language works237,207