Four-focus single-particle position determination in a confocal microscope

Lloyd M. Davis; Brian K. Canfield; James A. Germann; Jason K. King; William N. Robinson; Albert D. Dukes III; Sandra J. Rosenthal; Philip C. Samson; John P. Wikswo

doi:10.1117/12.842572

25 February 2010 Four-focus single-particle position determination in a confocal microscope

Lloyd M. Davis, Brian K. Canfield, James A. Germann, Jason K. King, William N. Robinson, Albert D. Dukes III, Sandra J. Rosenthal, Philip C. Samson, John P. Wikswo

Author Affiliations +

Proceedings Volume 7571, Single Molecule Spectroscopy and Imaging III; 757112 (2010) https://doi.org/10.1117/12.842572
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

We discuss the capabilities for sub-diffraction, single-nanoparticle position determination in a confocal one- or twophoton microscope with four-focus pulse-interleaved excitation and time-gated single-photon counting. As the technique is scalable to multiple detectors for multi-color observations, it can be used to find the separations of differently colored molecules over a distance range that is complementary to that achievable by FRET. Also, there is a possibility for improved spatial localization by using the nonlinearity of saturation of the excitation or by using the technique together with imaging of the point spread function. Applications of two experimental set-ups for four-focus fluorescence excitation for studies of quantum dots and single-particle manipulation and trapping are also discussed.

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Lloyd M. Davis, Brian K. Canfield, James A. Germann, Jason K. King, William N. Robinson, Albert D. Dukes III, Sandra J. Rosenthal, Philip C. Samson, and John P. Wikswo "Four-focus single-particle position determination in a confocal microscope", Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 757112 (25 February 2010); https://doi.org/10.1117/12.842572

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