No-lyse no-wash flow cytometry method for ex vivo study of human neutrophils
Bibliographic record
Abstract
Neutrophils are vital immune cells involved in immunity via their effector roles and death. While their rapid response to the environment is crucial for immune function, it also makes them difficult to study. Here, we provide a no-lyse no-wash (NLNW) flow cytometry protocol to quantify neutrophil viability and death in whole blood with minimal sample handling. Viability of neutrophils was measured by Annexin V/7AAD staining in whole blood at 0 and 20 h. Associated cost, sample volume, and neutrophil survival were compared with traditional density-gradient isolated neutrophils. The NLNW analysis of whole blood immediately after collection showed ∼85% viable neutrophils. After 20 h of incubation, viability dropped to ∼70%, yet was significantly greater compared with isolated neutrophils, where half the population (∼44%) entered early-stage apoptosis. The NLNW flow cytometry method is an efficient and cost-effective method to measure neutrophil viability in small volumes of whole blood with minimal sample handling.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot 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.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.004 | 0.001 |
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.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
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".