Protocol: Pooled dispensing procedure using Pico-Break™ with Cyto-Mine® Platforms
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Protocol Overview
Designed to allow for enrichment of antibody secreting cells (e.g. CHO cells lines, Plasma B cells, etc.), this protocol allows you to isolate cells from pool dispensed droplets on the Cyto-Mine® and Cyto-Mine® Chroma platforms using Pico-Break™.
Pooled dispensing involves dispensing all droplets contained within the dispense chamber into a single well of a 24-well plate rather than dispensing one droplet at a time into a single well of a 96- or 384-well plate.
Although it’s possible to isolate and dispense rare secretion events within a monoclonally encapsulated picodroplet, occasionally you may require several experimental repeats to obtain enough positive events or to run through all your isolate plasma B cells. In these situations, you can consider running an enrichment experiment using your Cyto-Mine® platform to increase the number of secreting cells obtained.
Enrichment allows you to isolate more rare events than a monoclonal experiment thereby increasing the number of clones or sequences that can be obtained. Depending on your experimental needs you may only require enrichment of your cell populations which can then be further processed downstream of the Cyto-Mine® platforms.
Through following this protocol developed by our in-house team, you can expect with high confidence to isolate secreting cells from a rare or low secreting population by using the Cyto-Mine® platforms to enrich and bulk dispensing droplets. The bulk dispensed droplets can then be broken using Pico-Break™ allowing for isolation of enriched cell populations for downstream workflows.
How it Works:
Step 1
Prepare your cell population and load onto the Cyto-Cartridge®
Step 2
Cell Encapsulation, Incubation, Gating, and Sorting on Cyto-Mine® or Cyto-Mine® Chroma
Step 3
Pooled dispensing of droplets into 24-well plate
Step 4
Gently break the picodroplets using Pico-Break™
Step 5
Use your enriched cell population for downstream processing
If you have any questions about how to get started or need our advice when carrying out this protocol, please contact our team at sales@fluidic.com and we’ll be happy to help!
The Next Generation: Cyto-Mine® Chroma
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Think Cyto-Mine®, but supercharged, enabling multiplexing and greater assay flexibility to fit your needs. It means that you can examine vastly greater numbers of cells — and isolate the most valuable ones— with unparalleled precision.
