Skip to Content
Merck
  • Visual barcodes for clonal-multiplexing of live microscopy-based assays.

Visual barcodes for clonal-multiplexing of live microscopy-based assays.

Nature communications (2022-05-19)
Tom Kaufman, Erez Nitzan, Nir Firestein, Miriam Bracha Ginzberg, Seshu Iyengar, Nish Patel, Rotem Ben-Hamo, Ziv Porat, Jaryd Hunter, Andreas Hilfinger, Varda Rotter, Ran Kafri, Ravid Straussman
ABSTRACT

While multiplexing samples using DNA barcoding revolutionized the pace of biomedical discovery, multiplexing of live imaging-based applications has been limited by the number of fluorescent proteins that can be deconvoluted using common microscopy equipment. To address this limitation, we develop visual barcodes that discriminate the clonal identity of single cells by different fluorescent proteins that are targeted to specific subcellular locations. We demonstrate that deconvolution of these barcodes is highly accurate and robust to many cellular perturbations. We then use visual barcodes to generate 'Signalome' cell-lines by mixing 12 clones of different live reporters into a single population, allowing simultaneous monitoring of the activity in 12 branches of signaling, at clonal resolution, over time. Using the 'Signalome' we identify two distinct clusters of signaling pathways that balance growth and proliferation, emphasizing the importance of growth homeostasis as a central organizing principle in cancer signaling. The ability to multiplex samples in live imaging applications, both in vitro and in vivo may allow better high-content characterization of complex biological systems.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Rapamycin, InSolution, ≥98%, 5 mM, inhibits mTOR and blocks activation of p70 S6 Kinase
Sigma-Aldrich
CDC25 Phosphatase Inhibitor I, BN82002, The CDC25 Phosphatase Inhibitor I, BN82002, also referenced under CAS 396073-89-5, controls the biological activity of CDC25 Phosphatase. This small molecule/inhibitor is primarily used for Phosphorylation & Dephosphorylation applications.
Sigma-Aldrich
Cdk2 Inhibitor III, The Cdk2 Inhibitor III, also referenced under CAS 199986-75-9, controls the biological activity of Cdk2. This small molecule/inhibitor is primarily used for Phosphorylation & Dephosphorylation applications.
Sigma-Aldrich
Cycloheximide solution, Ready-Made Solution, microbial, 100 mg/mL in DMSO, Suitable for cell culture