Biomek® FX-Based Core System Solution for Cell-Based Assay Screening of a Compound Library in a Core Lab Facility

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Noel Ruppert, Beckman Coulter, Inc.

To highlight the "plug and play" nature of the Biomek FX-based Core System, this article shares how a customer installed two microplate readers to cover the diversity of detection modes needed for the plethora of cell-based assays run on the system.

The user established this core facility in a university setting to run cell-based assays in high-throughput screening (HTS) mode. The system is used to automate mammalian cell-based and yeast genetic assays as well as high-throughput ELISA assays using cell extracts for compound screening of an existing library. On average, 95 microplates are used to screen 30,000 compounds for each assay. For cell-based assays, the appropriate cell line, depending on the application, is plated into 384-well plates and grown up in the Cytomat 6001 CO₂ incubator. On the day of the assay, compounds and reagents are added to the cultures and plates are read in the FlexStation II reader for kinetic assays or further incubated and read in the VICTOR3* multimode reader for detection of end-point assays. The Core System designed to provide this level of flexibility is shown (Figures A and B).

Figure A. Core System setup (Click the image for additional detail).

It is of additional interest to point out that the Biomek FX dual-bridge hybrid system was set up with an HDR tool on the Multi-Channel head and the Span-8 pipetting system on the other bridge.

The HDR tool was fitted with either fixed pins for compound transfers and plate replication. The challenge with using fixed tips or pins is the carryover of material leading to cross contamination of the compound source plates. The solution is to adequately clean the fixed tips or pins after each liquid transfer. The customer's solution was to implement a plasma-based cleaning device developed internally. The fixed tips or pins are then immersed in a high-frequency plasma field generated between two ceramic plates for cleaning. This approach results in reducing DMSO waste; long cycle times for pin cleaning and adequately removes sticky compounds so as to minimize contamination of source plates with dirty pins. The customer has also integrated a Branson ultrasonic bath onto the deck of the FX. The Branson bath was used in support of the pin cleaning and additionally can be used to help solubilize compounds that have formed a precipitate in solution.

The Span-8 pipetting function was used for hit picking and reformatting. The customer was trained to use a data file called the Hits File, which specifies wells to pick from the barcoded source plates. For additional information, see the "Custom Hit Picking Using Biomek FX Wizard for Span-8" in the June 2004 issue of T³ Update.

The customer also worked with the Project Management Team to optimize the performance of the Span-8 pipetting for low volumes. The goal was to pipette 800 nL with low percentage CV in an effort to preserve the compound library. This goal was met and the customer is very happy with the performance of the FX.

Figure B. Core System setup.

For Research Use Only; not for use in diagnostic procedures.

For comments or questions about T³ Update, please contact David Daniels, Ph.D., editor.

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