Olympus Scan^R Screening Station
| High Content Screening: General Information |
The Core Facility has an Olympus Scan^R screening station which allows automated image acquisition and data analysis of biological samples. The true power of the system lies in its ability to process large numbers of samples objectively, enabling their quantitative analysis.
What is High Content Screening (HCS)?
The Scan^R is one of a number of new machines recently developed in the area of high content screening or HCS. Other terms for this rapidly developing field include imaging cytometry, slide based cytometry, and automated microscopy. This type of instrument can combine the higher resolution and spatial localization abilities of a fluorescent microscope with the analysis possibilities and higher throughput reminiscent of flow cytometers. Cells or tissue sections located on slides, in chambers or in microtiter plates can be automatically scanned and than analysed for a number of different parameters. Although the total throughput is not as high as that of a standard flow cytometer it is still much higher than that of a typical microscope, allowing entire cell populations to be examined. Unlike flow cytometers, morphological information from cells and tissue sections can also be collected and used for analysis. The Scan^R allows analysis of this data through the use of histograms, dot plots and even population gating as in flow cytometry.
A typical multiwell experiment would consist of setting the number and relative positions of images to be taken in the selected wells, as well as the chosen colors/filter sets. Through the help of an autofocus function there is no need for further input from the operator until the scanning is completed, at which point it is then possible to carry out multi-parameter data analysis by choosing a combination of exclusion/inclusion criteria which can be applied to the saved images.
Sample Types
Sample types which can be investigated with the Scan^R include both adherent and non-adherent cells and tissue sections. A climate-controlled chamber is available so that living cells as well as fixed specimens can be examined. The environmental chamber also makes time lapse experiments possible. Additionally, our Scan^R system contains a disk scanning unit which could be useful for studying thicker specimens as well as during assay develpment.
Sample Templates
The Scan^R is very flexible with regard to what templates can be used for the samples. Plates and dishes with optical bottoms are recommended for the best quality images and lowest background.
- Microtiter plates (96- or 384-well)
- Chambered coverslips
- Standard slides
- Petri dishes
- Standard tissue culture plates
Excitation/Emission
The Scan^R comes equipped with a mixed-gas xenon-mercury lamp, which allows excitation of a broad range of fluorochromes. Currently filters are available for the following colors:
| Excitation | Emission | Examples of Dyes or Proteins which Could Be Used |
350 nm | 455 nm | DAPI, Hoechst |
| 458 nm | 477 nm | ECFP, Sytox Blue, BOPRO-1 |
488 nm | 508 nm | EGFP, Alexa 488, FITC, CY2, YOYO-1, DiOC6(3) |
514 nm | 527 nm | EYFP, TOTO-1, Oregon Green 514, Rhod-123, Bodipy-FL |
| 546 nm | 570 nm | Alexa 546, TRITC (Rhodamine), CY3, Propidium Iodide, RFP, PE |
594 nm | 615 nm | Texas Red, Alexa 594, CY 3.5 |
633 nm | 670 nm | Alexa 633, CY5, APC, TOTO-3, DRAQ-5 |
| 710 nm | 775 nm | CY7, Alexa 750 |
Additional filters/colors could be added if necessary. There is also the possibility of examining standard histologically stained specimens (brightfield) as well as of using differential interference contrast (DIC).
If you have any questions about possible labeling strategies for your experiment, we would be happy to discuss them with you or offer suggestions. We also have several staining/mounting protocols on hand which may be of interest.
Access
Only those investigators who have been previously trained to use the Scan^R Screening Station will be allowed to use it without an operator, otherwise it is necessary for a member of the Core Facility to assist you. For information about reserving time on the instrument, just contact one of us.
In addition to the Scan^R Screening Station itself, we also have a dedicated computer workstation and large capacity server for offline analysis and data storage.