Fundamentals ofCell-BasedAssays SRB5041 Andrei Leitão NEQUIMED-IQSC-USP email@example.com
Content • The mammalian cell. Main constituents & organization. Composition and structure. • Standard mammalian cell culture techniques. • Interrogating the biological system. Phenotypic and mechanism-based studies. • Networking in the cell. How do we account on this? Cell-signaling process. Cell fate: replication and death processes. Cell cycle, autophagy, apoptosis, necrosis. • Common assay platforms and techniques: • Colorimetric assays: Principles of the technique. Some dyes: methylene blue; phenol red; Tetrazolium salts. • Fluorometric assays: Principles of the technique and examples. • Flow cytometry: Principles of the technique and common assays. • Image-based techniques: bright field; fluorescence microscopy; electron microscopy. • Blotting the cell constituents: introduction to the southern, Western and eastern blots. • High-Throughput Screening (HTS) versus High-Content Screening (HCS) assays. • The next generation of assays: The impact of microscaled systems on cell-based assays. 3D cell culture, organ-on-a-chip & microfluidics. • Overview of chemical storage, handling and stability.