Z-polarized confocal microscopy
- 1 January 2001
- journal article
- Published by SPIE-Intl Soc Optical Eng in Journal of Biomedical Optics
- Vol. 6 (3) , 273-6
- https://doi.org/10.1117/1.1382610
Abstract
In light microscopy the transverse nature of the electromagnetic field precludes a strongly focused longitudinal field component, thus confining polarization spectroscopy and imaging to two dimensions (x,y). Here we describe a simple confocal microscopy arrangement that optimizes for signal from molecules with transition dipoles oriented parallel to the optic axis. In the proposed arrangement, we not only generate a predominant longitudinally (z) polarized focal field, but also engineer the detection scheme in such a way that in a bulk of randomly oriented molecules, the microscope's effective point-spread function is dominated by the contribution of those molecules that are oriented along the optic axis. Our arrangement not only implicitly allows for the determination of the orientation of transition dipoles of single molecules in three dimensions, but also highlights the contribution of z-oriented molecules in three-dimensional imaging.Keywords
This publication has 18 references indexed in Scilit:
- Three-dimensional orientation measurements of symmetric single chromophores using polarization microscopyNature, 1999
- Simultaneous Imaging of Individual Molecules Aligned Both Parallel and Perpendicular to the Optic AxisPhysical Review Letters, 1998
- Single-molecule spectroscopy with 27 fs pulses: Time-resolved experiments and direct imaging of orientational distributionsApplied Physics Letters, 1998
- An electromagnetic theory of imaging in fluorescence microscopy, and imaging in polarization fluorescence microscopyBioimaging, 1997
- Single molecules observed by immersion mirror objective. The orientation of terrylene molecules via the direction of its transition dipole momentChemical Physics Letters, 1997
- Fundamental reduction of the observation volume in far-field light microscopy by detection orthogonal to the illumination axis: Confocal theta microscopy (Optics Comm. 111 (1994) 536)Optics Communications, 1995
- Fundamental reduction of the observation volume in far-field light microscopy by detection orthogonal to the illumination axis: confocal theta microscopyOptics Communications, 1994
- Single Molecules Observed by Near-Field Scanning Optical MicroscopyScience, 1993
- 3‐D image formation in high‐aperture fluorescence confocal microscopy: a numerical analysisJournal of Microscopy, 1990
- Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic systemProceedings of the Royal Society of London. Series A. Mathematical and Physical Sciences, 1959