X-Y sample scanning stage and calibration method suitable for single-molecule detection

This paper describes the construction of a positioning device for sample scanning in the x and y directions suitable for single molecule fluorescence experiments. The mechanism uses a simple parallelogram flexure cut out of a single aluminum plate and two amplified piezoelectric actuators of the typ...

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Autores principales:	Treegate, K., Rasamessard, A., Osotchan, T., Hodak, J.H.
Formato:	JOUR
Materias:	Fluorescence spectroscopy Optical detection Scanning confocal microscopy Single molecule spectroscopy Aluminum plates Calibration method Correction techniques Cut out Diffraction-limited resolution Direct calibration Dwell time Fluorescent dyes Focus adjustment Imaging glass Line profiles Mechanical response Microscope objective Positioning devices Range images Scanning confocal microscopes Scanning device Scanning stages Single molecule fluorescence Single-molecule detection Target molecule Calibration Confocal microscopy Fluorescence Microscopes Piezoelectric actuators Scanning Molecules
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_09254005_v150_n1_p239_Treegate
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_09254005_v150_n1_p239_Treegate
record_format	dspace
spelling	todo:paper_09254005_v150_n1_p239_Treegate2023-10-03T15:46:12Z X-Y sample scanning stage and calibration method suitable for single-molecule detection Treegate, K. Rasamessard, A. Osotchan, T. Hodak, J.H. Fluorescence spectroscopy Optical detection Scanning confocal microscopy Single molecule spectroscopy Aluminum plates Calibration method Correction techniques Cut out Diffraction-limited resolution Direct calibration Dwell time Fluorescent dyes Focus adjustment Imaging glass Line profiles Mechanical response Microscope objective Optical detection Positioning devices Range images Scanning confocal microscopes Scanning confocal microscopy Scanning device Scanning stages Single molecule fluorescence Single molecule spectroscopy Single-molecule detection Target molecule Calibration Confocal microscopy Fluorescence Fluorescence spectroscopy Microscopes Piezoelectric actuators Scanning Molecules This paper describes the construction of a positioning device for sample scanning in the x and y directions suitable for single molecule fluorescence experiments. The mechanism uses a simple parallelogram flexure cut out of a single aluminum plate and two amplified piezoelectric actuators of the type used for microscope objective focus adjustment. A displacement range of 75 μm on each axis is obtained. The stage can be used to implement a sample scanning confocal microscope for single molecule spectroscopy applications using either inverted or up-right microscopes. Images with diffraction limited resolution can be obtained with this scanning stage. This is demonstrated by imaging glass beads labeled with the DY475 fluorescent dye and single rhodamine molecules. Micron sized range images of 256 × 256 pixels can be obtained with dwell times down to 0.5 ms/pixel. A novel direct calibration in which the mechanical response obtained from the line profiles for forward and reverse motion is used to account for the hysteresis of the stage. The target molecules are then located within the focus of the laser beam by using its corrected position. The performance of this scanning device and correction technique are demonstrated for the acquisition of fluorescence trajectories of individual rhodamine molecules. © 2010 Published by Elsevier B.V. Fil:Hodak, J.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_09254005_v150_n1_p239_Treegate
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Fluorescence spectroscopy Optical detection Scanning confocal microscopy Single molecule spectroscopy Aluminum plates Calibration method Correction techniques Cut out Diffraction-limited resolution Direct calibration Dwell time Fluorescent dyes Focus adjustment Imaging glass Line profiles Mechanical response Microscope objective Optical detection Positioning devices Range images Scanning confocal microscopes Scanning confocal microscopy Scanning device Scanning stages Single molecule fluorescence Single molecule spectroscopy Single-molecule detection Target molecule Calibration Confocal microscopy Fluorescence Fluorescence spectroscopy Microscopes Piezoelectric actuators Scanning Molecules
spellingShingle	Fluorescence spectroscopy Optical detection Scanning confocal microscopy Single molecule spectroscopy Aluminum plates Calibration method Correction techniques Cut out Diffraction-limited resolution Direct calibration Dwell time Fluorescent dyes Focus adjustment Imaging glass Line profiles Mechanical response Microscope objective Optical detection Positioning devices Range images Scanning confocal microscopes Scanning confocal microscopy Scanning device Scanning stages Single molecule fluorescence Single molecule spectroscopy Single-molecule detection Target molecule Calibration Confocal microscopy Fluorescence Fluorescence spectroscopy Microscopes Piezoelectric actuators Scanning Molecules Treegate, K. Rasamessard, A. Osotchan, T. Hodak, J.H. X-Y sample scanning stage and calibration method suitable for single-molecule detection
topic_facet	Fluorescence spectroscopy Optical detection Scanning confocal microscopy Single molecule spectroscopy Aluminum plates Calibration method Correction techniques Cut out Diffraction-limited resolution Direct calibration Dwell time Fluorescent dyes Focus adjustment Imaging glass Line profiles Mechanical response Microscope objective Optical detection Positioning devices Range images Scanning confocal microscopes Scanning confocal microscopy Scanning device Scanning stages Single molecule fluorescence Single molecule spectroscopy Single-molecule detection Target molecule Calibration Confocal microscopy Fluorescence Fluorescence spectroscopy Microscopes Piezoelectric actuators Scanning Molecules
description	This paper describes the construction of a positioning device for sample scanning in the x and y directions suitable for single molecule fluorescence experiments. The mechanism uses a simple parallelogram flexure cut out of a single aluminum plate and two amplified piezoelectric actuators of the type used for microscope objective focus adjustment. A displacement range of 75 μm on each axis is obtained. The stage can be used to implement a sample scanning confocal microscope for single molecule spectroscopy applications using either inverted or up-right microscopes. Images with diffraction limited resolution can be obtained with this scanning stage. This is demonstrated by imaging glass beads labeled with the DY475 fluorescent dye and single rhodamine molecules. Micron sized range images of 256 × 256 pixels can be obtained with dwell times down to 0.5 ms/pixel. A novel direct calibration in which the mechanical response obtained from the line profiles for forward and reverse motion is used to account for the hysteresis of the stage. The target molecules are then located within the focus of the laser beam by using its corrected position. The performance of this scanning device and correction technique are demonstrated for the acquisition of fluorescence trajectories of individual rhodamine molecules. © 2010 Published by Elsevier B.V.
format	JOUR
author	Treegate, K. Rasamessard, A. Osotchan, T. Hodak, J.H.
author_facet	Treegate, K. Rasamessard, A. Osotchan, T. Hodak, J.H.
author_sort	Treegate, K.
title	X-Y sample scanning stage and calibration method suitable for single-molecule detection
title_short	X-Y sample scanning stage and calibration method suitable for single-molecule detection
title_full	X-Y sample scanning stage and calibration method suitable for single-molecule detection
title_fullStr	X-Y sample scanning stage and calibration method suitable for single-molecule detection
title_full_unstemmed	X-Y sample scanning stage and calibration method suitable for single-molecule detection
title_sort	x-y sample scanning stage and calibration method suitable for single-molecule detection
url	http://hdl.handle.net/20.500.12110/paper_09254005_v150_n1_p239_Treegate
work_keys_str_mv	AT treegatek xysamplescanningstageandcalibrationmethodsuitableforsinglemoleculedetection AT rasamessarda xysamplescanningstageandcalibrationmethodsuitableforsinglemoleculedetection AT osotchant xysamplescanningstageandcalibrationmethodsuitableforsinglemoleculedetection AT hodakjh xysamplescanningstageandcalibrationmethodsuitableforsinglemoleculedetection
_version_	1782028146389811200

X-Y sample scanning stage and calibration method suitable for single-molecule detection

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