Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 120, Issue 26, Pages 6130-6138Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpcb.6b02153
Keywords
-
Categories
Funding
- Harvard MRSEC
- National Science Foundation [DMR-1420570]
- National Science Foundation under NSF [1541959]
- National Science Foundation Graduate Research Fellowship [DGE-1144152]
Ask authors/readers for more resources
A complete understanding of the cellular pathways involved in viral infections will ultimately require a diverse arsenal of experimental techniques, including methods for tracking individual viruses and their interactions with the host. Here we demonstrate the use of holographic microscopy to track the position, orientation, and DNA content of unlabeled bacteriophages (phages) in solution near a planar, functionalized glass surface. We simultaneously track over 100 individual lambda phages at a rate of 100 Hz across a 33 mu m x 33 mu m portion of the surface. The technique determines the in-plane motion of the phage to nanometer precision, and the height of the phage above the surface to 100 nm precision. Additionally, we track the DNA content of individual phages as they eject their genome following the addition of detergent-solubilized LamB receptor. The technique determines the fraction of DNA remaining in the phage to within 10% of the total 48.5 kilobase pairs. Analysis of the certain conditions, lambda phages move along the surface with their heads down and intermittently stick to the surface by their tails, causing them to stand up. Furthermore, we find that in buffer containing high concentrations of both monovalent and divalent salts, lambda phages eject their entire DNA in about 7 s. Taken together, these measurements highlight the potential of holographic microscopy to resolve the fast kinetics of the early stages of phage infection.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available