Construction and application of a single-molecule fluorescence microscope
Bibliographic record
Abstract
In this paper, we describe the construction, optimization, and testing of an epifluorescence microscope that has single-molecule (SM) resolution and sensitivity. The microscope makes use of a novel new type of wide-area charge-coupled device (CCD) photodetector with on-chip multiplier gain. Sensitivity and spatial resolution of the instrument are demonstrated by imaging individual Rhodamine 6G (R6G) molecules and characterizing their basic photophysical behaviour under a variety of imaging conditions. A simple, general method for calibrating the photodetector (correlating CCD counts with incident photons) using a highly attenuated laser beam is presented, and the performance of the photodetector is compared with that of other detectors commonly used in SM fluorescence imaging applications. We also demonstrate the versatility of the microscope system by characterizing the SM photophysical behaviour of several other fluorescent molecules, including bodipy-FL and the enhanced green fluorescent protein (EGFP). Key words: single molecule, fluorescence, microscopy, CCD camera, calibration, photobleaching.
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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".