Force Measurements on Single Molecular Contacts through Evanescent Wave Microscopy

Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave.

Fluorescence techniques for monitoring single-molecule dynamics in the vertical dimension currently do not exist. Here we use an atomic force microscope to calibrate the distance-dependent intensity decay of an evanescent wave. The measured evanescent wave transfer function was then used to convert the vertical motions of a fluorescent particle into displacement (SD = < 1 nm). We demonstrate th...

Ca2+-Triggered Peptide Secretion in Single Cells Imaged with Green Fluorescent Protein and Evanescent-Wave Microscopy

Green fluorescent protein fused to human chromogranin B or neuropeptide Y was expressed in PC12 cells and caused bright, punctate fluorescence. The fluorescent points colocalized with the endogenous secretory granule marker dopamine beta-hydroxylase. Stimulation of live PC12 cells with elevated [K+], or of permeabilized PC12 cells with Ca2+, led to Ca2+-dependent loss of fluorescence from neuri...

Trapping force and optical lifting under focused evanescent wave illumination.

A physical model is presented to understand and calculate trapping force exerted on a dielectric micro-particle under focused evanescent wave illumination. This model is based on our recent vectorial diffraction model by a high numerical aperture objective operating under the total internal condition. As a result, trapping force in a focused evanescent spot generated by both plane wave (TEM00) ...

Confocal microscopy on fluorescently labelled single DNA molecules and force - extension measurements on DNA carpets

Single Molecule Force Spectroscopy on Polysaccharides by Atomic Force Microscopy

Recent developments in piconewton instrumentation allow the manipulation of single molecules and measurements of intermolecular as well as intramolecular forces. Dextran filaments linked to a gold surface were probed with the atomic force microscope tip by vertical stretching. At low forces the deformation of dextran was found to be dominated by entropic forces and can be described by the Lange...