Abstract
Åderförkalkning är en sjukdom i de stora artärerna och en viktig bakomliggande orsak till hjärtinfarkt och stroke. Flera olika musmodeller har utvecklats för att underlätta studier av den molekylära och cellulära patofysiologin hos denna sjukdom. I detta manuskript beskriver vi specifika metoder för kvantifiering och karakterisering av aterosklerotiska lesioner i murina aorta sinus och stigande aorta. Fördelen med detta förfarande är att det ger en noggrann mätning av den tvärsektionsarea och den totala volymen av lesionen, som kan användas för att jämföra aterosklerotisk progression mellan olika behandlingsgrupper. Detta är möjligt genom användning av klaffbladen såsom en anatomisk landmärke, tillsammans med noggrann inställning av snittvinkel. Vi beskriver också grundläggande färgningsmetoder som kan användas för att börja att karaktärisera aterosklerotisk progression. Dessa kan modifieras ytterligare för att undersöka antigener av specifik interest till forskaren. De beskrivna teknikerna gäller allmänt för ett stort antal befintliga och nyskapade kost och genetiskt-inducerade modeller av aterogenes.
Materials
Name | Company | Catalog Number | Comments |
LDLR-deficient mice (D2.129S7(B6)-Ldlrtm1Her/J) | The Jackson Laboratory | stock #002207 | |
Formalin | Sigma-Aldrich | HT5011 | |
Histology Molds | Ted Pella Inc | 27195 | |
Paraffin | Paramat | 19286-10 | |
Coated slides | Fisher | 12-550-15 | |
Solvent containers | Tissue Tek II | 4457 | |
Microtome | Leica | RM22S5 | |
Water bath | VWR | 80086-982 | |
Microscope | Olympus | BX41TF | |
Camera | Olympus | DP72 | |
ImageJ | NIH | www.rsbweb.nih.gov/ij/ | |
Processor | Sakura Finetek | VIP6-A1 | |
Processing cassettes | VWR | 18000-134 | |
Preassure Cooker | Nordic Ware | ||
Coverslip | Fisher | 12-545-M | |
PBS | Sigma-Aldrich | P3813 | |
Microtome blade | Thermo Scientific | 30-518-35 | |
Antifade mounting medium | Sigma-Aldrich | F-4680 | |
Xylene mounting medium | Sigma-Aldrich | 44581 | |
Aquous mounting medium | Sigma-Aldrich | I1161 | |
Mac-3 anbody | BD Pharmingen | 553322 | |
DAPI | Sigma-Aldrich | D9542 | |
F4/80 antibody | Abcam | Ab6640 | |
Alpha actin antibody | Santa Cruz Biotechnology | SC-32251 | |
Mayer’s Hematoxylin | Sigma-Aldrich | E4382 | |
Eosin Y | Sigma-Aldrich | MHS16 | |
DAB (3,3'-Diaminobenzidine) | Dako | K3468 |
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