Whole-Larva Cryosectioning and Immunolabeling of<i>Drosophila</i>Larvae
Bibliographic record
Abstract
Resolution in microscopy—the shortest distance between which objects can be distinguished from each other—is crucial for our ability to view details of biological samples. The theoretical resolution limit of light microscopy is 200 nm in the x,y -plane. Using stacks of x,y images, 3D reconstructions of the z -plane of a specimen can be achieved. However, because of the nature of light diffraction, the resolution of the z -plane reconstitutions is closer to 500–600 nm. Peripheral nerves of the fruit fly Drosophila melanogaster consist of several thin layers of glial cells surrounding the underlying axons. The size of these components can be well under the resolution of z -plane 3D reconstructions, thus making it difficult to determine details of coronal views through these peripheral nerves. Here, we describe a protocol to obtain and immunolabel 10-μm cryosections of whole third-instar larvae of the fruit fly Drosophila melanogaster . Cryosectioning the larvae using this method converts visualization of coronal sections of the peripheral nerve into the x,y -plane and brings the resolution down from 500–600 nm to 200 nm. Theoretically, this protocol can also be used with some modifications to obtain cross sections of other tissues.
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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 | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.002 | 0.001 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.001 | 0.003 |
| Insufficient payload (model declined to judge) | 0.012 | 0.010 |
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".