First author Xiaoming Lu recently relocated to the RTP area and is looking for a lab to call home as well! If you’re in the market for someone with extensive cell and molecular biology skills that also knows how to code in python send me an email and I’ll put you in touch.
Posts by Birtwistle Lab
That is related to previous work on MuSIC (multiplexing using spectral imaging and combinatorics). pubmed.ncbi.nlm.nih.gov/30339749/ and pubmed.ncbi.nlm.nih.gov/37463472/ .
Why do this? We thought such barcodes could be useful for lineage tracing or genetic perturbations. Also, we think it could be possible to generate many more barcodes (even into the millions) by creating tandem fluorescent protein fusions that have unique spectra due to FRET.
As proof-of-concept we made 2-way combinations of 18 fluorescent proteins spanning UV-to-IR. This makes 153 possible barcodes. We showed that they are largely uniquely resolvable, with a few exceptions, in human cells via spectral flow cytometry (4-laser Aurora from CytekBio).
We had a lot of fun with @markblenner over the past several years developing and experimentally demonstrating the concept of fluorescent protein barcodes using spectral readouts, recently published: pubs.acs.org/doi/full/10....
Additionally, although it is not in this paper, the company developed 48 well gels that double the migration distance, greatly increasing molecular weight resolution and adding more options for our users.
Compared to the original pre-print, we added data showing MW ranges resolvable with different acrylamide % gels (overall 10 to 250 kDa), limit-of-detection equivalency with a matched regular western, and a dual color blot (thank you to Azure and their solid instruments!).
Thanks to reviewers along the way, NIGMS/NIH for small business grant support, XLerateHealth and 3PhaseSC for critical support and guidance, and recent SCRA support to help bring the technology to market.