Transnational Call

SMALL MOLECULE SCREENING

 

Phenotypic or targeted cellular screen based on high-content analysis

Abstract

In this call, the IMTM-HTS facility offers long term scientific expertise in live-cell based screening and high-content analysis of reporter or stably transfected cells for monitoring of cytoplasmatic, nuclear or nucleolar targets and processes; e.g., DNA damage and repair in fixed and live cells. The HTS system consists of three robotic arms, automatic incubators, liquid handlers, readers for fluorescence, luminescence, absorbance or ionizing radiation, X-Ray irradiator, and confocal microscopes. An essential part of such screening is available image analysis by advanced software algorithms at our computational cluster.

Description of technology offered under this call

In this call, the IMTM-HTS facility offers long term scientific expertise in live-cell based screening and high-content analysis. The IMTM facility can provide assays directed towards a wide range of targets involved in vital biological processes such as DNA damage response or the cell cycle progression. The assays are based on imaging of reporter or stably transfected cells and monitoring of diverse processes such as signal accumulation in a specific compartment or cytoplasmic to nuclear translocation. Our instrumentation enables live-cell imaging and time-lapse microscopy followed by a high-content analysis of acquired images.

IMTM has a modular HTS platform that allows testing in BSL3 and BSL2+ environments, screening in combination with ionizing radiation (X-rays), mass spectrometry, high-content analysis and other technologies. The screening assays provide hits for downstream drug research and development. The HTS/HCA screening platform is based on a state-of-art robotic system provided by HighResBiosolutions Ltd. The system consists of three robotic arms, automated incubators, liquid handlers for microliter and nanoliter volumes, sealers, de-sealers, centrifuges and readers for fluorescence, luminescence, absorbance and ionizing radiation. Also integrated with the system are wide-field or spinning disc confocal microscopes (Operetta, Yokogawa CV7000) equipped with software tools for image analysis and data evaluation. The IMTM screening facility can offer assays for screening of biologically active compounds and anticancer agents, assays for identification of adenosine receptor agonists and antagonists as well as mapping of the most significant signaling pathways (e.g., Hedgehog, Wnt, p53, KRAS, RAGE) via phosphorylation and reporter assays. In addition to mammalian cells, IMTM laboratories are fully equipped for the screening of antimicrobial activity in G+/G- strains including drug-resistant bacteria, mycobacterial strains, yeasts, filamentous fungi, viruses and parasites under BSL2 and at limited capacity under BSL3 conditions.

Number of compounds screened

100,000 (full commercial set) of the EU-OPENSCREEN ERIC compound library

Time frame for screen up to validated hits

10 months

Machine/Methods offered under this call

  • HighRes Biosciences robotic platform for ultra-high-throughput testing of biological activity of small molecules
  • HighRes Biosolutions Inc. robotic system for storage of chemical libraries and preparation of test panels of these chemicals
  • X-Ray irradiation device
  • Yokogawa CV7000 Voyager high-throughput cellular imaging and discovery system
  • PerkinElmer Operetta imaging system
  • PerkinElmer Columbus system for image data analysis and management, optionally the Mathlab based scripts for image analysis; Image-J or CellProfiler available.
  • PerkinElmer EnVision HTS multilabel plate reader
  • PerkinElmer MicroBeta radioactivity and luminescence counter

Services provided under this call

  • Validation of the assay provided by the user to ensure robustness (Yokogawa CV7000 Voyager, image analysis and data management using Columbus).
  • The miniaturization of the test into 384-well or 1536-well plates (Viability and response assays, Yokogawa CV7000 Voyager, EnVision).
  • Pilot screen using 5,000 compounds from the EU-OS compound library (Yokogawa CV7000 Voyager, image analysis and data management using Columbus).
  • Analysis of pilot screen data (Columbus, TIBCO Spotfire).
  • Full screen with the remaining 95,000 compounds of the EU-OS compound library (Yokogawa CV7000 Voyager, Columbus).
  • Data analysis and subsequent selection of primary hits (Columbus, TIBCO Spotfire).
  • Hit picking for hit validation, preparation of secondary screening plates with the hits, preparation of source plates for dose-response (Tecan EVOware).
  • Hit confirmation, generation of dose-response curves using the primary assay and IC50 calculation (Yokogawa CV7000 Voyager, Columbus, Dotmatics Studies).
  • Further hit evaluation will be done in collaboration with the “applicant” in relation to the studied processes.

Note

Please note that this project includes the possibility of re-screening during potential chemical optimization under the medicinal chemistry call starting in 2021 (tentative date).

Prerequisites for applicants

The applicant is expected to have a project with clearly defined objectives. The applicant needs to show preliminary data confirming the reproducibility of the biological assay or demonstrate a measurable change in phenotype of the cell line used. Before the transfer of any cell line to our facility, the applicant has to prove that the cell line is free of mycoplasma. The applicant should effectively communicate or be temporarily present on the screening site in order to facilitate the adaptation to automated screening and provide key bespoke reagents.

As specifics of the assay transfer procedure may vary between partner sites, the applicant and the individual sites will agree on the appropriate steps and logistics together.

 

Partner site


IMTM

Institute of Molecular and Translational Medicine

Faculty of Medicine and Dentistry

Palacky University Olomouc

Hnevotinska 1333/5

779 00 Olomouc, Czech Republic

https://www.imtm.cz/