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Fundamentals of Cell-Based Assays

Fundamentals of Cell-Based Assays. SRB5041 Andrei Leitão NEQUIMED-IQSC-USP andleitao@iqsc.usp.br. Content. • The mammalian cell. Main constituents & organization. Composition and structure. • Standard mammalian cell culture techniques.

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Fundamentals of Cell-Based Assays

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  1. Fundamentals ofCell-BasedAssays SRB5041 Andrei Leitão NEQUIMED-IQSC-USP andleitao@iqsc.usp.br

  2. 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.

  3. References – some samples

  4. References – some samples

  5. Schedule

  6. Schedule

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