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Abstract:

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.

Registro:

Documento: Artículo
Título:X-Y sample scanning stage and calibration method suitable for single-molecule detection
Autor:Treegate, K.; Rasamessard, A.; Osotchan, T.; Hodak, J.H.
Filiación:Department of Physics, Faculty of Science, Mahidol University, Rama 6 Rd, Phayathai, Bangkok 10400, Thailand
INQUIMAE-Departamento de Quimica Inorganica, Analitica y Quimica Fisca, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria, AR-1428 Buenos Aires, Argentina
Palabras clave: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
Año:2010
Volumen:150
Número:1
Página de inicio:239
Página de fin:246
DOI: http://dx.doi.org/10.1016/j.snb.2010.07.008
Título revista:Sensors and Actuators, B: Chemical
Título revista abreviado:Sens Actuators, B Chem
ISSN:09254005
CODEN:SABCE
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09254005_v150_n1_p239_Treegate

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Citas:

---------- APA ----------
Treegate, K., Rasamessard, A., Osotchan, T. & Hodak, J.H. (2010) . X-Y sample scanning stage and calibration method suitable for single-molecule detection. Sensors and Actuators, B: Chemical, 150(1), 239-246.
http://dx.doi.org/10.1016/j.snb.2010.07.008
---------- CHICAGO ----------
Treegate, K., Rasamessard, A., Osotchan, T., Hodak, J.H. "X-Y sample scanning stage and calibration method suitable for single-molecule detection" . Sensors and Actuators, B: Chemical 150, no. 1 (2010) : 239-246.
http://dx.doi.org/10.1016/j.snb.2010.07.008
---------- MLA ----------
Treegate, K., Rasamessard, A., Osotchan, T., Hodak, J.H. "X-Y sample scanning stage and calibration method suitable for single-molecule detection" . Sensors and Actuators, B: Chemical, vol. 150, no. 1, 2010, pp. 239-246.
http://dx.doi.org/10.1016/j.snb.2010.07.008
---------- VANCOUVER ----------
Treegate, K., Rasamessard, A., Osotchan, T., Hodak, J.H. X-Y sample scanning stage and calibration method suitable for single-molecule detection. Sens Actuators, B Chem. 2010;150(1):239-246.
http://dx.doi.org/10.1016/j.snb.2010.07.008