13 research outputs found
Radial Distribution Function for Semiflexible Polymers Confined in Microchannels
An analytic expression is derived for the distribution of the
end-to-end distance of semiflexible polymers in external potentials
to elucidate the effect of confinement on the mechanical and statistical
properties of biomolecules. For parabolic confinement the result is exact
whereas for realistic potentials a self-consistent ansatz is developed, so that
is given explicitly even for hard wall confinement. The
theoretical result is in excellent quantitative agreement with fluorescence
microscopy data for actin filaments confined in rectangularly shaped
microchannels. This allows an unambiguous determination of persistence length
and the dependence of statistical properties such as Odijk's deflection
length on the channel width . It is shown that neglecting the
effect of confinement leads to a significant overestimation of bending
rigidities for filaments
Real-time imaging of single DNA molecules with fluorescence microscopy.
A fluorescence microscopy technique was used to image the dynamics of individual DNA molecules. Lambda, calf thymus, cosmid (circular), and T4 DNA were studied with the fluorescent dye acridine orange. Experiments with DNAase I were conducted, and the results indicate that these observations correspond to DNA molecules. The results of experiments with circular DNA provide strong evidence that these were single DNA molecules. Molecules were observed free in solution or attached to a glass or copper surface at one or several points. The Brownian motion of these molecules was observed, indicating that DNA in solution exists in a partially supercoiled state. Some molecules appeared stretched and were attached to the surface by their termini; the lengths of these molecules were measured. Such molecules also exhibited elastic behavior upon breaking. The power of this technique is demonstrated in images of cosmid DNA molecules, catenanes, and DNA extending from T4 phage particles. These results suggest immediate applications to molecular biology, such as examining the dynamics of protein-DNA interactions. Areas of ongoing research are discussed