Enhanced sensitivity in dark-field microscopy by optimizing the illumination angle

Taylor, Michael A. and Bowen, Warwick P. (2013) Enhanced sensitivity in dark-field microscopy by optimizing the illumination angle. Applied Optics, 52 23: 5718-5723. doi:10.1364/AO.52.005718


Author Taylor, Michael A.
Bowen, Warwick P.
Title Enhanced sensitivity in dark-field microscopy by optimizing the illumination angle
Journal name Applied Optics   Check publisher's open access policy
ISSN 1559-128X
2155-3165
Publication date 2013-08
Year available 2013
Sub-type Article (original research)
DOI 10.1364/AO.52.005718
Volume 52
Issue 23
Start page 5718
End page 5723
Total pages 6
Place of publication Washington DC, United States
Publisher Optical Society of America
Collection year 2014
Language eng
Abstract Dark-field microscopy is a well-known technique used to exclude the bright background of unscattered photons from a measurement. We show that by choosing an appropriate illumination angle, the background of unwanted scattered light can also be suppressed. The collected flux of scattered photons is calculated in the Mie scattering regime for various particle sizes and objectives over a range of illumination angles. In the case that the dark-field measurement is limited by background scattering, we find that the sensitivity can be improved by lowering the objective numerical aperture. The collected photon flux is calculated for an exemplary dark-field microscopy experiment in which lipid granules were studied within yeast cells. Our model suggests that the signal-to-noise ratio was over three-orders-of-magnitude higher than it would have been with an equivalent bright-field setup.
Keyword Light-Scattering
Biological Cells
Refractive-Index
Resolution
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: School of Mathematics and Physics
Official 2014 Collection
 
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