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Protocol for analyzing γδ T cells from the vagina of diet-induced obese mice using single-cell RNA sequencing and flow cyto ... 2 Before you begin 2 Institutional Permissions 2 Preparation of herpes simplex virus (HSV)-2 2 Titration of HSV-2 stock 3 Induction of high fat diet-induced obesity mouse model 4 Key resources table 5 Materials and equipment 6 2-Mercaptoethanol stock solution 6 Collagenase IV-stock solution 6 DNase I-stock solution 6 Hyaluronidase-stock solution 7 Dispase II-stock solution 7 HSV-2 incubation media 7 Dispase II mixture 8 Anti-γδTCR antibody coating solution 9 Magnetic sorting media 9 MPA solution 9 Step-by-step method details 10 1: Infection of HSV-2 10 2: Isolation of cells from mouse vaginal tissue 11 3: Analysis of immune cells obtained from mouse vaginal tissue by single-cell RNA sequencing 13 4: Analysis of immune cells obtained from mouse vaginal tissue by flow cytometry 15 5: In vitro expansion and flow cytometric analysis of murine γδ T cells 17 Expected outcomes 21 Quantification and statistical analysis 21 Limitations 21 Troubleshooting 21 Problem 1 21 Potential solution 21 Problem 2 21 Potential solution 21 Problem 3 22 Potential solution 22 Prob

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nonfiction