Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy

Paul A. Dalgarno, Heather Dalgarno (nee Campbell), Aurelie Putoud, Robert Lambert, Lynn Paterson, David C. Logan, David P. Towers, Richard J. Warburton, Alan H. Greenaway

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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 ~10 nm z position resolution by parameterising the images with an image sharpness metric. © 2010 Optical Society of America.

Original languageEnglish
Pages (from-to)877-884
Number of pages8
JournalOptics Express
Volume18
Issue number2
DOIs
Publication statusPublished - 18 Jan 2010

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Dalgarno, P. A., Dalgarno (nee Campbell), H., Putoud, A., Lambert, R., Paterson, L., Logan, D. C., Towers, D. P., Warburton, R. J., & Greenaway, A. H. (2010). Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy. Optics Express, 18(2), 877-884. https://doi.org/10.1364/OE.18.000877