Agnostic Tracking: Nanoscale, High Bandwidth, 3D Particle Tracking for Biology

Public Deposited

Last Modified

Creator

Desai, Kalpit V.
- Affiliation: School of Medicine, UNC/NCSU Joint Department of Biomedical Engineering

Abstract

The ability to detect biological events at single molecule level provides unique insights in the field of biophysics. Back-focal-plane laser interferometry is a promising technique for single-molecule-scale, 3D position measurements at rates far beyond the capability of video. I present an in-situ calibration method for the back-focal-plane, low-power (non-trapping) laser interferometry. The software-based technique does not rely on any a priori model or calibration knowledge; hence the name Agnostic. The technique is sufficiently fast and non-invasive that the calibration can be performed on the fly, without interrupting or compromising the on-going experiment. The technique can be applied to track 3D, long range motion (up to 100 um) of a broad variety of microscopic biological objects. The spatiotemporal resolution achieved is of the order of a few nanometers and tens of microseconds. Three biological applications enabled by the technique are presented: firstly, a prototype of an oscillating-bead high-bandwidth frequency-response analyzer for biology, based on Agnostic Tracking as implemented in our custom-built 3D Magnetic Force Microscope (3DFM); secondly, a magnetic-force study that revealed a previously-unknown anchoring-dependent nonlinear response of a cellular membrane; last, a rheological study that revealed a novel grouping of motion characteristics of individual vesicles diffusing inside live cytoplasm.

Date of publication

DOI

Resource type

Rights statement

Advisor

Language

Access right

Date uploaded

Relations

Thumbnail	Title	Date Uploaded	Visibility	Actions
	Agnostic tracking : nanoscale, high bandwidth, 3D particle tracking for biology	2019-04-10	Public	Download