An Electroporation Cytometry Protocol for Live-Cell, Fluorescence Microscopy Using U2 OS Cell Culture
Bibliographic record
Abstract
Pulsed electric fields (PEFs) have a wide range of applications in medical research and clinical applications. A key area of research focuses on electroporation (reversible or irreversible). Reversible electroporation has been used for several decades for transferring molecules through cell membranes such as plasmid DNA, typically referred to as gene electrotransfer (GET). Conversely, irreversible electroporation has become a popular technique in cancer treatment, providing a non-toxic alternative by permanently rupturing cells using PEFs. This can be combined with various types of drugs or reagents to enhance the effect which has become known as electrochemotherapy (ECT). However, despite the broad success in practical applications, further supporting technology and research is required to improve current techniques. To address this, an electroporation cytometry system was developed to support live-cell PEF experimentation with the ability to perform long-term fluorescence microscopy. This was in combination with a stably expressing FUCCI(CA)5 U2-OS cell line for assessing changes in the cell cycle in response to PEF exposure. In doing so, up to 30 h timelapse microscopy was achieved, providing real-time changes in cell activity based on the FUCCI(CA)5 reporter. As a result, phase-specific changes were quantifiable for downstream analysis.
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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) | 0.000 | 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 | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| 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".