TY - GEN
T1 - In vivo three-dimensional superresolution fluorescence tracking using a double-helix point spread function
AU - Lew, Matthew D.
AU - Thompson, Michael A.
AU - Badieirostami, Majid
AU - Moerner, W. E.
PY - 2010
Y1 - 2010
N2 - The point spread function (PSF) of a widefield fluorescence microscope is not suitable for three-dimensional superresolution imaging. We characterize the localization precision of a unique method for 3D superresolution imaging featuring a double-helix point spread function (DH-PSF). The DH-PSF is designed to have two lobes that rotate about their midpoint in any transverse plane as a function of the axial position of the emitter. In effect, the PSF appears as a double helix in three dimensions. By comparing the Cramer-Rao bound of the DH-PSF with the standard PSF as a function of the axial position, we show that the DH-PSF has a higher and more uniform localization precision than the standard PSF throughout a 2 μm depth of field. Comparisons between the DH-PSF and other methods for 3D superresolution are briefly discussed. We also illustrate the applicability of the DH-PSF for imaging weak emitters in biological systems by tracking the movement of quantum dots in glycerol and in live cells.
AB - The point spread function (PSF) of a widefield fluorescence microscope is not suitable for three-dimensional superresolution imaging. We characterize the localization precision of a unique method for 3D superresolution imaging featuring a double-helix point spread function (DH-PSF). The DH-PSF is designed to have two lobes that rotate about their midpoint in any transverse plane as a function of the axial position of the emitter. In effect, the PSF appears as a double helix in three dimensions. By comparing the Cramer-Rao bound of the DH-PSF with the standard PSF as a function of the axial position, we show that the DH-PSF has a higher and more uniform localization precision than the standard PSF throughout a 2 μm depth of field. Comparisons between the DH-PSF and other methods for 3D superresolution are briefly discussed. We also illustrate the applicability of the DH-PSF for imaging weak emitters in biological systems by tracking the movement of quantum dots in glycerol and in live cells.
KW - Fluorescence microscopy
KW - Quantum dot
KW - Single-particle tracking
KW - Superresolution
KW - Three-dimensional microscopy
UR - http://www.scopus.com/inward/record.url?scp=77951668923&partnerID=8YFLogxK
U2 - 10.1117/12.842608
DO - 10.1117/12.842608
M3 - Conference contribution
AN - SCOPUS:77951668923
SN - 9780819479679
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Single Molecule Spectroscopy and Imaging III
T2 - Single Molecule Spectroscopy and Imaging III
Y2 - 23 January 2010 through 25 January 2010
ER -