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High Throughput And High Content Screening PowerPoint Presentation
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High Throughput And High Content Screening

High Throughput And High Content Screening

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High Throughput And High Content Screening

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  1. High Throughput And High Content Screening

  2. What Is HTS ? Biochemical target  Test many compounds  Compounds with desired effect on target = Hit  OR    Hit directly used as chemical probe to study target Hit provides structure for chemists to begin developing a drug or other product

  3. HTS Technology • Robotics • Miniaturization • Sophisticated Assay Chemistry • Sophisticated Software and Database

  4. Evotec uHTS System • Opera High Content Detector • Plate:vision detector for fluorescence (also fluorescence polarization & fluorescence resonance energy transfer), luminescence, absorbance • 2 CO2 incubators • 4 low volume pipetting systems for 384- and 96-well plates (multidrops, Cy-Bi well, Cy-By drop) • Plate washer • Centrifuge • Piercer • Plate storage, barcode readers, lid handlers, transport systems

  5. plate::vision detector CCD camara Xenon lamp light source Zeiss optics Built in algorithm to correct for spherical lens aberration 4 filter sets Excitation filter Emission filter Dichroic filter

  6. Opera • 4 lasers • 1 xenon lamp • 4 CCD cameras • 96- and 384-well plates • Acapella image analysis software

  7. Software Systems • Genedata • Assay Analyzer • Condeseo • Integrated with Opera Acapella image analysis software • Accelyrs Accord • HTS • With Pipeline Pilot • Ohio Third Frontier Computing Systems

  8. HTS • Output is • QUALITY DATA

  9. Standard Screening Procedure • 1o HTS  hits • Triplicate assay  confirmed HTS hits • 10 point dose-response assay  EC50

  10. HTS Assay Examples • Receptor-binding • GPCR • Enzyme • protease, kinase, phosphatase, lipase, others • Bacterial growth • Cell-based reporter gene • Cell growth, cell viability • Cytochrome P450 inhibition • Protein binding • Protein production by cells • ELISA

  11. HIGH-THROUGHPUT SCREENING HIGH-THROUGHPUT SCREENING Examples of Assays:

  12. Results from Enzyme Inhibition Screen • 11,500 compounds screened • Normalized results shown • Each tile shows 1 plate of data • Right: • Color Scale for Normalization • Distribution of compound % Activity

  13. Results from 1 plate in Enzyme Inhibition Screen

  14. High Content Screening With the Evotec Opera 96-, 384-, and 1536-well plates Up to ~ 70 images per well in 384-well plate

  15. Internalisation Assay Example Image

  16. Internalisation Assay Example Image Analysis

  17. Internalisation Assay Example Opera External Data output Excel spreadsheet spots / cell cell area nuclei area

  18. High Throughput & High Content Screening Services • Quality Data • Experienced Scientists • State-of-the Art Equipment

  19. Contacts • Sandra Nelson • High Throughput & High Content Screening • sandra.nelson@uc.edu • 558-2372 • Ruben Papoian • Drug Discovery • ruben.papoian@uc.edu • 558-0516 • Charles McOsker • Business Development & External Relations • charles.mcosker@uc.edu • 558-2569