# Ultra High-Throughput Immune Cell Analysis ![[Pasted image 20240106205352.png|350]] 1. Single cell emulsification: Flow focusing device combines 2. **Cell lysis and mRNA capture:** The VH & VL mRNA from each cell will be *co-localized* on the Poly(dT) beads present as the emulsion droplet only contains a single cell (aka native pairing information is retained). 3. **Beads are recovered** and original emulsions are broken but we ensure that native pairing is not disrupted and the mRNA present remains associated with their original beads. 4. **Emulsion Linkage RT-PCR:** Beads are reemulsified for cDNA synthesis following their recovery. The isolated nature of this reaction allows us to recovery physically linked heavy & light chain fragments via overlap extension (linkage) RT-PCR. --- - Supports **BOTH** *high-throughput sequencing* **AND** *functional analysis* (via cloning into display systems) - Fragment antigen-binding (Fab) is a little bit *more thermally stable* and *more native* than most single-chain fragment variable (scFV) ![[Pasted image 20240106203740.png|400]]