Electroporation of Adherent Cells In Situ for the Introduction of Nonpermeant Molecules
Bibliographic record
Abstract
Electroporation has been used for the introduction of DNA ( 1 – 4 ), proteins ( 5 – 7 ), and various nonpermeant drugs and metabolites into cultured mammalian cells ( 8 , 9 ), as described in this and other volumes ( 10 , 11 ). Most electroporation techniques for adherent cells involve the delivery of the electrical pulse while the cells are in suspension ( 5 , 12 ). However, the detachment of these cells from their substratum by trypsin or EDTA can cause metabolic alterations that may lead to cell death or increase the cells’ sensitivity to additional damaging agents ( 13 ). The efficient incorporation of proteins without cell damage is an especially crucial requirement, since for most proteins of interest and contrary to DNA, no convenient large-scale method exists for the selection of viable from damaged cells or cells where no protein introduction took place after electroporation. Therefore, for studies using adherent cells, it is imperative to deliver the pulse while the cells are attached to their solid substratum. This has been achieved using a variety of methods ( 14 , 15 ). In this chapter, we describe a technique where cells are grown and electroporated on a glass surface coated with electrically conductive, optically transparent indium-tin oxide. This coating allows the direct visualization of the electroporated cells and offers the possibility of ready examination owing to their extended morphology. Moreover, it is very durable, inert, nontoxic to the cells, and promotes excellent cell adhesion and growth. An added advantage is the fact that, unlike many plastics, indium-tin oxide does not exhibit any spontaneous fluorescence, making the examination of the electroporated cells under a fluorescent microscope possible. These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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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 | 0.000 | 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 | 0.001 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.003 | 0.002 |
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".