UiO is offering a suspension cell-based screening project using a flow cytometry based analysis with automated sample preparation. Potential readouts for the screen include basic cell characteristics like size and granularity, assessment of recombinant proteins (e.g. GFP), cell surface markers and intracellular staining like cell cycle analysis.
UiO has a strong background in immunology, cancer immunology and cell signaling analysis based on flow cytometry methods and is equipped with bio safety level 2 cell culture labs to handle primary human cells as well as established human cell lines. It has established a pipeline for the automated preparation of cell samples for flow cytometry in 384 format based on a Beckman Coulter Biomek i7 with an integrated plate centrifuge.
This setup is capable of running different protocols including preparation steps like cell fixation and permeabilization using various reagents like paraformaldehyde, saponine and methanol to prepare samples for extra and intracellular staining with agents including conjugated antibodies and DNA staining agents. Depending on volume requirements, staining reagents can be transferred using either the Biomek i7, liquid dispensers or an Echo acoustic liquid handler for volumes down to 2.5 nL. Samples will be analyzed on an Intellicyt (Sartorius) iQue screener PLUS flow cytometer equipped with 3 lasers (13 color channels). Possible readouts for the flow cytometry screen include, but are not limited to basic cell characteristics like size and granularity, expression of recombinant proteins (e.g. labelled with GFP), cell surface markers (e.g. CD molecules) and DNA stain for cell cycle analysis.
100,000 (full commercial set) of the EU-OPENSCREEN ERIC compound library
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).
The prerequisite starting point to qualify for access is the availability of an established bioassay and associated key bespoke reagents, developed at lab-scale by the successful applicant. Criteria to define anestablished bioassay are compatibility with 96-well format (assay volume up to 150 µL), demonstrated successful flow cytometry based analysis by the applicant (e.g. successful detection of recombinant proteins by flow cytometry and not a different method like western blotting) and the availability of positive and negative controls.
Key bespoke reagents include the assay ready cells or particles that will be analyzed by flow cytometry (e.g. for expression analysis of recombinant proteins, the applicant has to provide the transfected cells, cell culture can be carried out on site) and special staining reagents like commercial kits and antibodies.
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.
Cell-based assays using complex cellular systems
drug discovery, small molecules, high-throughput screening, cell-based assays
June-01-2019 (20:00 CET)
September-30-2019 (20:00 CET)
Scientific / Technical / Methods contact:
Chemical Biology Platform, Centre for Molecular Medicine Norway (NCMM),
Faculty of Medicine,
University of Oslo (UiO)
Forskningsparken Gaustadalléen 21
0349 Oslo, Norway