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

Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy.

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  • 1School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, UK.


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.

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