Based on the CellRaft Technology, the CytoSort™ Array comprises an elastomeric microwell array with each well of the array containing separately releasable “CellRaft”. Precision soft lithography methods are used to obtain uniform dimensions across the array. The CellRafts are formed in a second fabrication step by condensing in each microwell a polymer containing paramagnetic particles.
The CytoSort Array is currently available in two sizes:
– 10,000 microwells each microwell having a dimension of 200 x 200 microns
– 40,000 microwells each microwell having a dimension of 100 x 100 microns.
Cells seeded on the CytoSort Array are localized on the CellRaft surface and remain viable. The array is housed in a petri dish-sized 65 mm cassette with a reservoir for various media, buffers or other reagents, making it familiar to the biological research community.
Once cells settle and adhere to the CellRafts, the CytoSort Array can be screened to identify target cells or colonies. The CytoSort Array can be imaged by brightfield, fluorescence and confocal microscopy. Each CellRaft within the CytoSort Array has a unique address, describing its row and column position within the array. Individually released CellRafts can be transferred to analysis tubes for biochemical or molecular analysis.
The imaging, release, and collection of CellRafts from a CytoSort Array is accomplished using either the automated AIR System or the CellRaft System for Inverted Microscopes.
The CytoSort Array has been specifically designed for use in the automated CellRaft AIR System. A unique code is embedded in the array that is read by the AIR System to identify the array format and materials.
The user can image the CytoSort Array either on or off the AIR System. If used on a standard inverted microscope the user can manually generate a list of addresses in a Microsoft Excel-style spreadsheet. The spreadsheet can be directly imported into the “Real Time” run mode of the AIR System operating software.
When used in the AIR System, the CytoSort Array is held in place on the deck of the instrument by an adapter. The adapter is keyed so that the Array can be moved in and out of the adapter and still maintain proper orientation for multiple imaging runs. The clear border of the CytoSort Array allows the AIR System to efficiently scan every microwell position.
Once the array is positioned on the adapter, the entire assembly can be placed on the left side of the deck of the AIR™ System. Alternatively, the adapter can be placed on the deck first followed by placing the CytoSort Array into the adapter. The right position is intended to house a 96-well SBS format plate for collecting the CellRafts. The AIR™ System employs the same proprietary needle-based release and magnet-based collection strategy as the manual CellRaft System for Inverted Microscopes.
The image below is taken from the AIR System user interface and shows a single cell stained on a CytoSort Array. RED: SYTO 17 (cytoplasm), GREEN: Alexa 488 phalloidin (actin), BLUE: Hoechst (nucleus).