Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy

Opt Express. 2010 Jan 18;18(2):877-84. doi: 10.1364/OE.18.000877.

Abstract

A conventional microscope produces a sharp image from just a single object-plane. This is often a limitation, notably in cell biology. We present a microscope attachment which records sharp images from several object-planes simultaneously. The key concept is to introduce a distorted diffraction grating into the optical system, establishing an order-dependent focussing power in order to generate several images, each arising from a different object-plane. We exploit this multiplane imaging not just for bio-imaging but also for nano-particle tracking, achieving approximately 10 nm z position resolution by parameterising the images with an image sharpness metric.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement / instrumentation*
  • Imaging, Three-Dimensional / instrumentation*
  • Microscopy / instrumentation*
  • Nanoparticles / ultrastructure*
  • Nanotechnology / instrumentation*
  • Refractometry / instrumentation*
  • Reproducibility of Results
  • Sensitivity and Specificity