用流式细胞仪分析脂肪细胞 .ppt

Adipocytes are large and fragile 50–200 μm Easy to be broken, when the flow rate (pressure) is large. Contaminants Stromal/vascular cells Aggregates of adipocytes Free lipid droplets Other debris * Adipocytes are large and fragile 50–200 μm ——using flow cytometry with larger internal diameter of the fluidics (150 to 250μm). Easy to be broken——control Flow pressures to 5~10 psi, slow but not broken. Good for isolation. * Contaminants Stromal/vascular cells Aggregates of adipocytes Free lipid droplets Other debris ——Exclusion of Contaminants (4 steps) * Analysis and Isolation of Adipocytes by Flow Cytometry Majka SM, Miller HL, Helm KM, et al. Analysis and Isolation of Adipocytes by Flow Cytometry. Methods in enzymology. 2014;537:281-296. doi:10.1016/B978-0-12-411619-1.00015-X. * Background Aim of the research Difficulty Solution What are adipocytes (fat cells)?Major cells in the adipose tissue. * Lipid droplet Two kinds of adipocytes in human * Prospects of adipocyte biology Lipid store Inhibit eating Increase metabolism Leptin * Prospects of adipocyte biology Adipokines Involved in energy metabolism Lipid store * Prospects of adipocyte biologySpecifically from adipose tissue  Lipid store Adipokines Energy metabolism * Prospects of adipocyte biologyAdipokines play an significant role in biological function. * Flow Cytometry (FCM) Principle diagram * Volume, Morphological plexity of cells, Total DNA/RNA content, Apoptosis, Cell viability, ... ... ... ... * A method for analysis and isolation free-floating adipocytes by Flow Cytometry. Analysis Flow cytometry (FCM) Confocal laser scanning microscope (CLSM) Isolation Fluorescence-activated cell sorting (FACS) * Analysis Flow cytometry (FCM) Confocal laser scanning microscope (CLSM) Beckman Coulter--MoFlo? XDP Analysis rate:100,000 events/sec. Sort rate: 70,000 events/sec. * A method for analysis and isolation free