1 / 20

High Throughput And High Content Screening

High Throughput And High Content Screening. 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.

aubrey-cash
Download Presentation

High Throughput And High Content Screening

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  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

More Related