title: Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy.
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.
authors.affiliation: School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, UK. P.A.Dalgarno@hw.ac.uk
authors.forename: Paul A
authors.initials: PA
authors.lastname: Dalgarno
authors.forename: Heather I C
authors.initials: HI
authors.lastname: Dalgarno
authors.forename: Aurélie
authors.initials: A
authors.lastname: Putoud
authors.forename: Robert
authors.initials: R
authors.lastname: Lambert
authors.forename: Lynn
authors.initials: L
authors.lastname: Paterson
authors.forename: David C
authors.initials: DC
authors.lastname: Logan
authors.forename: David P
authors.initials: DP
authors.lastname: Towers
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Warburton
authors.forename: Alan H
authors.initials: AH
authors.lastname: Greenaway
mesh_terms: D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D008853:Microscopy / Q000295:instrumentation*; D053758:Nanoparticles / Q000648:ultrastructure*; D036103:Nanotechnology / Q000295:instrumentation*; D012031:Refractometry / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity