Live-Cell Movies

The following movies are from: I. Golding. “Single-Cell Studies of Phage λ: Hidden Treasures Under Occam’s Rug.” Annual Review of Virology 2016 [PDF].

Cell lysis following λ infection, as seen under phase-contrast illumination. The cell is infected at t = 0 and lyses at t ≈ 80 min.

Video by Louis McLane and Samuel Skinner; Golding lab.

Tracking the postinfection decision. Phage capsids were labeled using gpD-YFP. At t = 0, two cells are each infected by a single phage (green), and one cell is infected by three phages. The two cells infected by single phages go into the lytic pathway, as indicated by the intracellular production of new phage capsids (green). The cell infected by three phages goes into the lysogenic pathway, as indicated by the activity of a PRE-mCherry reporter (red). Eventually, the lytic pathway results in cell lysis, whereas the lysogenic cell continues to grow and divide.

Video reproduced from Zeng L, Skinner SO, Zong C, Sippy J, Feiss M, Golding I. 2010. “Decision making at a subcellular level determines the outcome of bacteriophage infection.” Cell 141:682–91, copyright 2010, with permission from Elsevier.

PR activity during lysogen induction. In the reporter strain, mRNA from the PR promoter is labeled using MS2-GFP (green), whereas the genome locus where the reporter resides is labeled using TetR-mCherry (red). The cells are λ lysogens. At t = 20 min, mitomycin C is added, leading to transient derepression of PR and induction of the dormant prophage. The prophage used is lysis deficient; hence, the cells do not lyse at the end of the induction process.

Video by Jing Zhang, Louis McLane, and Samuel Skinner; Golding lab.

Spatiotemporal dynamics of phage (red) and bacterial (green) genome loci inside the infected Escherichia coli cell during the lysogenic pathway. The phage genome is labeled using mCherry-ParB. In the same cell, the bacterial attB site is detected using an orthogonal ParB system and a GFP.

Video by Louis McLane and Samuel Skinner; Golding lab. For more information see Tal A, Arbel-Goren R, Costantino N, Court DL, Stavans J. 2014. “Location of the unique integration site on an Escherichia coli chromosome by bacteriophage lambda DNA in vivo.” PNAS 111:7308–12

A single gpD-YFP-labeled λ phage (green) diffusing near and on the Escherichia coli cell (red). Two examples are shown sequentially.

Video by Eli Rothenberg, Samuel Skinner, and Ido Golding. For more information, see Rothenberg E, Sepulveda LA, Skinner SO, Zeng L, Selvin PR, Golding I. 2011. “Single-virus tracking reveals a spatial receptor-dependent search mechanism”. Biophys. J. 100:2875–82