Abstract
Single-particle fluorescence microscopy of association/dissociation is required for analysis of biological assembly reactions. Toward achieving this goal, Wang et al. (J. Microsc., 2004, 213, 101-109) used molten agarose to concentrate thermally diffusing particles in a thin zone of solution next to the surface of a coverglass (plane of concentration). The present study details the first real-time, single-particle analysis of the association/dissociation of thermally diffusing particles in the plane of concentration. The test particles were procapsids of bacteriophage λ (radius = 31 nm). Quantification of thermal motion was developed and used to determine whether co-diffusing particles were bound to each other. The data are explained by (1) the presence of a molten agarose-generated barrier that is 93-155 nm from the coverglass surface, and (2) nonrandom orientation of procapsid dimers in the plane of concentration.
Original language | English (US) |
---|---|
Pages (from-to) | 83-92 |
Number of pages | 10 |
Journal | Journal of Microscopy |
Volume | 217 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2005 |
Keywords
- Alexa
- Association/dissociation
- Bacteriophage
- Capsid
- Confined thermal motion
- Dye
- Fluorescence microscopy
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Histology