Immunohistochemistry UCP1 and Lipid Droplets Images
Notice bibliographique
Résumé
Hematoxylin and eosin (H&E) staining protocol and lipid droplet analysis BAT was collected from naked mole-rats treated in normoxia (n = 12), or 1 or 3 hrs of hypoxia (n = 11 each) and samples prepared as described previously1. Briefly, interscapular BAT (iBAT) was dissected, cleaned to remove any white adipose, muscle or connective tissues, and then fixed in 10% formalin overnight. iBAT was then ethanol dehydrated and stored in 70% ethanol prior to paraffin embedding. Paraffin embedded tissue was sectioned to the largest surface area and used for H&E staining. After H&E staining, Mirax Viewer Image software (version 1.6) was used to analyze the sections in a ZEISS-MIRAX Midi Slide scanning system (Zeiss Microimaging, Oberkochen, Germany, and 3DTech, Budapest, Hungary) and digital images were acquired at 20x magnification. Images were extracted using Aperio ImageScope software (version 12.3.3; Leica Biosystems), and then were processed using Zeiss software ZEN 3.2 (Zen Lite; Carl Zeiss Canada Ltd. Toronto, Canada). ROIs were carefully selected in each image using the rectangular selective tool bar in the ZEN 3.2 software, making sure to avoid blood vessels and edges of the tissue in the ROI selected. All the ROI images produced were converted to a TIFF format. Then the images were analyzed by a blinded researcher using FIJI (ImageJ; NIH). The range thresholding that was applied across all replicate images in all conditions was 200-255 and the range of lipid droplet area analyzed was between 1-1000µm². The scale of each image processed was calibrated by adding a scale according to the ROI analyzed. Finally, the total lipid droplet area of each image was measured, and a percent lipid area was calculated using ImageJ and as described elsewhere2. Immunohistochemistry staining of interscapular BAT of NMR IHC staining was performed on formalin fixed paraffin embedded tissue sections using the Leica Bond™ system using a modification of protocol F that eliminates the post primary step when using rabbit antibodies on rat tissue. Sections were pre-treated using sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes. The sections were then incubated using a 1:1000 dilution of Rabbit UCP1 for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. Slides were then stained using DAB as the chromogen, counterstained with Hematoxylin, mounted and cover slipped. UCP1 positive cell manual counting of IHC-stained NMR interscapular BAT sections IHC UCP1 DAB-stained images for three conditions (Normoxia, Hypoxia 1hr & 3hrs) were converted into TIFF format and then opened with FIJI (ImageJ; NIH). For each replicate, 2 sections of 2000x2000 pixels were taken and then further analyzed. With FIJI (ImageJ; NIH), Plugin>Analyze>Cell Counter> Cell counter was selected to count the UCP1 positive and negative cells manually. The images were counted manually by selecting UCP1 positive cells and negative cells and then the data was exported to an excel spreadsheet. The UCP1 positive cells were divided by the total number of cells in each section to obtain the percentage of UCP1 positive cells which was graphed. Statistical analysis was made including the ANOVA one-way test and a t-test multiple comparison.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,001 | 0,001 |
| Méta-épidémiologie (sens large) | 0,001 | 0,001 |
| Bibliométrie | 0,003 | 0,002 |
| Études des sciences et des technologies | 0,001 | 0,001 |
| Communication savante | 0,001 | 0,001 |
| Science ouverte | 0,001 | 0,001 |
| Intégrité de la recherche | 0,001 | 0,002 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,026 | 0,007 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».