A fourdimensional microscope for single molecule studies in living cells using astigmatic axial resolution enhancement
Bibliographic record
Abstract
<p>An important part of life is to react to the environment and adapt to it. All living cells rely on chemical cascades that form signal pathways, from stimulus to transcription events at the DNA. To know more about these pathways we need tools to visualise the signals.</p> <p>Here is outlined developmental and experimental work on a bespoke microscope system primarily for fast single molecule tracing of proteins in living eukaryote cells. Both hardware and software has been developed. Frame rates and exposure times have been pushed to match the speed of freely diffusing particles in living cells. An EMCCD camera with an electronic gain of 300x and a 5 ms exposure time at 200 frames per second enables this.</p> <p>An astigmatic axial resolution enhancement approach has been implemented to allow tracking with high precision of single fluorescent proteins in real-time in three dimensions in a comparatively large volume. The high resolution in three spatial dimensions and one temporal aim to meet these demands. Software to extract the additional axial information has been developed that can determine axial displacement with 38 nm precision.</p> <p>The microscope system has proven it can find and track small clusters of YPet fluorescent protein in <em>E. coli</em> at short exposure times.</p>
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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 | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.001 |
| 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".