Mechanical stability of a microscope setup working at a few kelvins for single-molecule localization

[Display omitted] ► Localization precision of the image of a point source is free from diffraction. ► Prerequisite for the precision is stability of the sample and objective at 1.5K. ► We developed a rigid imaging unit to make image of scattering of a sample bead. ► The centroid of the scattering im...

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Bibliographic Details
Published in:	Chemical physics Vol. 419; pp. 246 - 249
Main Authors:	Hinohara, Takuya, Hamada, Yuki I., Nakamura, Ippei, Matsushita, Michio, Fujiyoshi, Satoru
Format:	Journal Article
Language:	English
Published:	Elsevier B.V 01-06-2013
Subjects:	Fluorescence imaging Localization precision Low temperature Single molecule Fluorescence imaging Localization precision Single molecule Low temperature
Online Access:	Get full text
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Summary:	[Display omitted] ► Localization precision of the image of a point source is free from diffraction. ► Prerequisite for the precision is stability of the sample and objective at 1.5K. ► We developed a rigid imaging unit to make image of scattering of a sample bead. ► The centroid of the scattering image of a bead was determined for 800 images. ► The standard deviation of the 800 centroids measured in 17min was 0.85nm. A great advantage of single-molecule fluorescence imaging is the localization precision of molecule beyond the diffraction limit. Although longer signal-acquisition yields higher precision, acquisition time at room temperature is normally limited by photobleaching, thermal diffusion, and so on. At low temperature of a few kelvins, much longer acquisition is possible and will improve precision if the sample and the objective are held stably enough. The present work examined holding stability of the sample and objective at 1.5K in superfluid helium in the helium bath. The stability was evaluated by localization precision of a point scattering source of a polymer bead. Scattered light was collected by the objective, and imaged by a home-built rigid imaging unit. The standard deviation of the centroid position determined for 800 images taken continuously in 17min was 0.5nm in the horizontal and 0.9nm in the vertical directions.
ISSN:	0301-0104
DOI:	10.1016/j.chemphys.2013.02.024