Development of a Microscope-based High Content Screening Platform

Urban I. Liebel
Cell Biology and Biophysics
EMBL-Heidelberg
Meyerhofstrasse 1
69117 Heidelberg
Germany
telephone: 49 6221 387232
fax: 49 6221 387306
email: liebel@embl-heidelberg.de
prestype: Poster
presenter: Urban I. Liebel

Urban I. Liebel, Jeremy C. Simpson and Rainer Pepperkok

The identification of large numbers of novel cDNAs about which no functional information exists, represents an enormous resource which should be exploited. Tagging each of these coding sequences with the green fluorescent protein (GFP), then expressing these fusion proteins in living cells allows a number of microscopic assays to be performed, which in turn generate functional data about these molecules. To this end, we have developed a fully automatic microscope which is integrated with a robotic liquid handling station.

We have written software which allows full control over objectives (10x, 20x, 40x, 63x), excitation/emission and neutral density filters. An image acquisition module for the cooled 12-bit CCD camera and a module to control the precision xy-stage are also integrated into the software. Assays have been developed in a 96-well format using thin glass-bottomed plates. A software autofocus has been developed to compensate for the well bottom roughness, and a fast hardware autofocus is under construction. This will decrease the screening time by a factor of ten. We are currently integrating this platform into a standard LIMS (Laboratory Information Management System) software environment in order to screen multiple plates without user intervention.

Different datamining modules, also developed in our lab, analyse the images taken by the microscope. The software can determine which cells are transfected with the GFP-tagged cDNAs, and quantify the fluorescence from multiple channels. The integration of these components allow us to rapidly carry out hundreds of individual experiments on these novel cDNAs which contribute the first information about these novel proteins.