Birtwistle Lab

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.

Fluorescence Multiplexing with Spectral Imaging and Combinatorics - PubMed Ultraviolet-to-infrared fluorescence is a versatile and accessible assay modality but is notoriously hard to multiplex due to overlap of wide emission spectra. We present an approach for fluorescence ...

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).

Genetically Encoded Fluorescence Barcodes Allow for Single-Cell Analysis via Spectral Flow Cytometry Genetically encoded, single-cell barcodes are broadly useful for experimental tasks such as lineage tracing or genetic screens. For such applications, a barcode library would ideally have high diversi...

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.

A 96-Well Polyacrylamide Gel for Electrophoresis and Western Blotting Western blotting is a stalwart technique for analyzing specific proteins and their post-translational modifications. However, it remains challenging to accommodate more than ∼10 samples per experiment...

Our high-throughput, 96-well western blot paper is out! pubs.acs.org/doi/10.1021/... If anyone is interested, contact Blotting Innovations–www.blottinginnovations.com or @BetterBlotting!