Internal standard application strategies in mass spectrometry imaging by desorption electrospray ionization mass spectrometry
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Bibliographic record
Abstract
RATIONALE: We developed a model case study to evaluate three internal standard (IS) application strategies (methods I-III) using the psycholeptic phenobarbital (PB) and the isotopically labelled IS phenobarbital-D5 (PB-D5) from in vitro dosed tissues of the golden apple snail (Pomacea diffusa) by desorption electrospray ionization mass spectrometry imaging (DESI-MSI). METHODS: In method I, the IS was deposited as microspots on top of 10 μm thick snail tissues; in method II, a thin IS film was applied; and in method III, the IS was spiked into the DESI solvent spray. DESI-MSI analyses were performed using a Thermo LTQ mass spectrometer equipped with a custom-built DESI source and two-dimensional moving stage. PB (m/z 231) and PB-D5 (m/z 236) were monitored in selected ion monitoring mode between m/z 227 and 239. RESULTS: The analytical performance of two IS strategies (methods I and II) in DESI-MSI was evaluated based on an intra- and inter-day precision assay, an accuracy assessment, and statistical analysis. In the inter-day DESI-MSI assay, method I exhibited better precision (6.5%-7.4%) than method II (10.7%-17.6%) between 10 and 100 ng/μL. In the accuracy assessment, PB quality controls of 75 ng/μL were back-calculated as 71 ± 4 and 83 ± 9 ng/μL, resulting in relative errors of -5% and 11% for methods I and II, respectively. Method III did not work under the experimental design and was not evaluated. CONCLUSIONS: Three IS application strategies were investigated and compared for a routine quantitative DESI-MSI approach. Methods I and II were not statistically significantly different as shown by a Bland-Altman plot, suggesting that these two methods can be used interchangeably. However, method III requires further research for future quantitative DESI-MSI analyses.
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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 | 0.000 | 0.005 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.001 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 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 it