High-throughput Single Cell Motility Analysis using Nanowell-in-Microwells
Bibliographic record
Abstract
Abstract Cell motility is important to many biological processes including cancer, immune response, and tissue repair. Conventional assays measure bulk cell motility, potentially overlooking important heterogeneity and missing important high motility subpopulations. Here, we introduce a high-throughput single-cell motility assay using nanowell-in-microwell plates to precisely track single cell position and analyze their migratory trajectories. By physically confining individual cells in nanowells, we eliminate cell-cell interactions and simplify cell segmentation and tracking. Using this platform, we characterized the motility of single cells across different culture conditions to identify distinct motility phenotypes. Single-cell trajectory analysis revealed pronounced directional persistence, with cells predominantly maintaining their direction of travel and trajectory along nanowell boundaries. Additionally, our approach facilitates the generation of labeled image datasets suitable for AI models to rapidly identify cell motility phenotypes from single-cell images. Together, our platform provides a robust, scalable method to analyze cell motility phenotypes and migration behavior at single-cell resolution. Significance Statement A novel single-cell motility assay that uses confinement in nanowell-in-microwell to enable high-throughput profiling of single cell motility phenotypes.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.001 | 0.002 |
| 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) | 0.000 | 0.000 |
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 teacher head, 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".