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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2010
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09254005_v150_n1_p239_Treegate http://hdl.handle.net/20.500.12110/paper_09254005_v150_n1_p239_Treegate |
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paper:paper_09254005_v150_n1_p239_Treegate2023-06-08T15:51:16Z X-Y sample scanning stage and calibration method suitable for single-molecule detection Hodak, Jose Hector 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. 2010 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09254005_v150_n1_p239_Treegate 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 Hodak, Jose Hector 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. |
| author |
Hodak, Jose Hector |
| author_facet |
Hodak, Jose Hector |
| author_sort |
Hodak, Jose Hector |
| 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 |
| publishDate |
2010 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09254005_v150_n1_p239_Treegate http://hdl.handle.net/20.500.12110/paper_09254005_v150_n1_p239_Treegate |
| work_keys_str_mv |
AT hodakjosehector xysamplescanningstageandcalibrationmethodsuitableforsinglemoleculedetection |
| _version_ |
1768541618481135616 |