Information for users:

Investigators using the facility are required to sign an agreement with the facility prior to use stating their willingness to submit a copy of their data to the facility for use internally in analyzing compound activity across screens and to facilitate collaborations with other facility users. Users of the facility are charged a nominal fee to compensate the facility for consumable use and the screening facility staff's time. After completion of a successful preliminary screen, users are assisted in completing a full screen of our current compound collection. Operation of the liquid handling equipment, bar-coding software and equipment, and compound handling and storage is performed solely by the screening facility staff. The screening facility staff assists investigators in use of the plate reader for assay detection as well as aiding investigators in the performance of all steps of their screen. To get started click here.

Non-Screening Services Offered:
I. Multi-Plex Cytotoxicity Assays
II. Wound Healing and Chamber Migration Assays

Follow the links below to find out how to use SMSF services:
I. Starting a screen of Chemical Libraries at SMSF
II. Performing a Preliminary Screen
III. Scheduling a preliminary screen or full screen
IV. Analyzing your preliminary screen data
V. Requesting a cytotoxicity assay at SMSF

Multi-Plex Cytotoxicity Assays:
The SMSF will perform cytoxicity assays on cancer cell lines as a service to anyone who wants to assess cytotoxicity of lead compounds. Compounds will be tested in duplicate at 5 concentrations around the estimated IC50 of the compounds. % Inhibition will be determined for each concentration tested and IC50s will be determined if 50% Inhibition is observed at any concentration tested. Cells are plated, treated with test compound, incubated for 72 hours and then assayed with a multi-plexed assay for cell viability. The simultaneous read outs of cytotoxicity include: Calcein AM (InVitrogen, Inc.), Cell-Titer glo (Promega, Inc), and Mitotracker Red (InVitrogen,inc.). (back to the top)
Cancer Cell Lines Maintained at the Facility:

(back to the top)

II.Wound Healing Assays and Chamber Migration Assays:

The screening facility now offers 2 assay services to identify small molecules that inhibit migration and invasion. The first assay is a modified boyden chamber assay, the BD Biosciences FluoroBlok Chamber Migration Assay. Compound and media are plated in the bottom plate of the migration chamber. Cells and compound are plated in the top plate of the migration chamber. Chemoattractant (5% FBS) was added at either time 0 or after a 1 hour pre-incubation of cells with compound. Cells are allowed to migrate for 24 hours and then stained with a fluorescent viable cell marker and read from the bottom in the Safire II plate reader. Data is corrected for background migration by subtracting the average counts of cells treated with 25 uM Blebbistatin, a potent inhibitor of migration. IC50s are determined using XLFit 4.2.

The second assay service offered is a quantitative wound healing assay. Cells are plated at high cell density per well of a 96-well plate and allowed to attach. A consistent and uniform wound of approximately 1.58 mm wide and 2.5 mm long is greated using a 96 mechanical floating pin tool modified to scratch a monolayer on the surface of a plate:

Cells are grown for 72 hours, stained with a viable cell marker and read on the plate reader 2X. The first read measures the total fluorescence over the whole well. The second read measures the fluorescence in the “X-Line” – the path of the wound. The wound ratio is calculated by dividing the X-line read by the whole well read. The data is corrected for background by subtracting the lowest X-line reading from the data. (back to the top)

Some Papers of Interest to SMSF Users:
Ross Stein "A new model for drug discovery - meeting our societal obligation"
Stuart L. Schreiber, "The small-molecule approach to biology: Chemical genetics and diversity-oriented organic synthesis make possible the systematic exploration of biology"
K. Balakin "DMSO Solubility and Bioscreening"

I. Starting a screen of chemical libraries at SMSF (back to the top)
Please read and fill out the following forms and return to the SMSF. When the SMSF receives your application you will be contacted to set up a preliminary meeting.
Application for Screening
Screening application (Cover Letter)
Keck-UWCCC Screening Agreement

II. Performing a Preliminary Screen (back to the top)

The purpose of the preliminary screen is to optimize the liquid handling and detection equipment, determine the best concentration of compound to use and to determine the hit rate. During the course of the preliminary screen we will optimize the volume of assay, concentration of compound stock to use, liquid handling steps and optimize the sensitivity of the read-out on the plate reader.

III. Scheduling a preliminary screen or full screen (back to the top)
View The Schedule (UW users only):
(Non-university users can contact us to schedule screening time)

Typically the SMSF schedule is open for the next week. If you would like to view the SMSF calendar:

1. Log onto my.wisc.edu
2. Sign into my WiscCal
3. Click on View Agendas in the calendar toolbar
4. Search for "Noel Peters" under name search
5. Highlight "Noel Peters" and click view agenda
6. To request time on the schedule contact us.

IV. Analyzing your preliminary screen data (back to the top)
After completion of your preliminary screen we ask that you analyze your data and determine either a %CV or Z-Factor. If your hit rate is greater than 0.3% or there is high variability observed we will repeat the preliminary screen to determine the optimal conditions before performing the full screen. For more information about analyzing screening data please read this paper:
A Simple Statistical Parameter For Use in Evaluation and Validation of High Throughput Assays.

V. Requesting a cytotoxicity assay at SMSF (back to the top)
If you would like request a cytotoxicity assay at SMSF, please submit the following application:
Application for Cytotoxicity Assays

Contact Us

© 2003 The University of Wisconsin
This file was updated 10/07/2003