TY - GEN
T1 - Single-molecule orientation measurements with a quadrated pupil
AU - Backer, Adam S.
AU - Backlund, Mikael P.
AU - Lew, Matthew D.
AU - Diezmann, Alexander R.
AU - Sahl, Steffen J.
AU - Moerner, W. E.
PY - 2014
Y1 - 2014
N2 - We present a means of measuring the dipole orientation of a fluorescent, rotationally fixed single molecule (SM), using a specially designed phase mask, termed a "quadrated pupil," conjugate to the back focal plane of a conventional widefield microscope. In comparison to image-fitting techniques that infer orientation by matching simulations to defocused or excessively magnified images, the quadrated pupil approach is more robust to minor modeling discrepancies, defocus, and optical aberrations. Precision on the order of 1°-5° is achieved in proofof- concept experiments for both azimuthal (φ) and polar (θ) angles. Since the phase mask is implemented on a liquid-crystal spatial light modulator (SLM) that may be deactivated without any mechanical perturbation of the sample or imaging system, the technique may be readily integrated into conventional imaging studies.
AB - We present a means of measuring the dipole orientation of a fluorescent, rotationally fixed single molecule (SM), using a specially designed phase mask, termed a "quadrated pupil," conjugate to the back focal plane of a conventional widefield microscope. In comparison to image-fitting techniques that infer orientation by matching simulations to defocused or excessively magnified images, the quadrated pupil approach is more robust to minor modeling discrepancies, defocus, and optical aberrations. Precision on the order of 1°-5° is achieved in proofof- concept experiments for both azimuthal (φ) and polar (θ) angles. Since the phase mask is implemented on a liquid-crystal spatial light modulator (SLM) that may be deactivated without any mechanical perturbation of the sample or imaging system, the technique may be readily integrated into conventional imaging studies.
KW - Fluorescence microscopy
KW - Optical Fourier processing
KW - Single-molecule microscopy
KW - Super-resolution
UR - http://www.scopus.com/inward/record.url?scp=84901754166&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84901754166&partnerID=8YFLogxK
U2 - 10.1117/12.2042097
DO - 10.1117/12.2042097
M3 - Conference contribution
AN - SCOPUS:84901754166
SN - 9780819498632
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Single Molecule Spectroscopy and Superresolution Imaging VII
PB - SPIE
T2 - Single Molecule Spectroscopy and Superresolution Imaging VII
Y2 - 1 February 2014 through 2 February 2014
ER -