论文信息 - Resolution enhancement in a light-sheet-based microscope (SPIM).

Resolution enhancement in a light-sheet-based microscope (SPIM).

Light-sheet-based microscopy [single-plane illumination microscope (SPIM)] performs very well at low numerical apertures. It complements conventional (FM), confocal (CFM), and two-photon fluorescence microscopy (2hnu-FM) currently used in modern life sciences. Lateral and axial SPIM point spread function (PSF) extents are measured by using fluorescent beads to determine the 3D resolution. The results are compared with values derived from an analytical theory and numerical simulations. The discrepancies are found to be less than 5%. The axial extent of a SPIM-PSF (10x/0.3 W) is approximately 5.7 microm. This value is almost a factor of 2 smaller than in CFM, more than 2.5 times smaller than in FM, and more than three times smaller than in 2hnu-FM. SPIM outperforms 2hnu-FM and FM, while CFM has a better axial resolution at NAs above 0.8.

Christoph J. Engelbrecht | E. Stelzer | Christoph J Engelbrecht | Ernst H Stelzer

[1] C. McCutchen. Generalized Aperture and the Three-Dimensional Diffraction Image , 1964 .

[2] Jan Huisken,et al. Multiple imaging axis microscopy improves resolution for thick-sample applications. , 2003, Optics letters.

[3] F. Del Bene,et al. Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[4] Steffen Lindek,et al. Fundamental reduction of the observation volume in far-field light microscopy by detection orthogonal to the illumination axis: confocal theta microscopy , 1994 .

[5] Farooq Azam,et al. Thin laser light sheet microscope for microbial oceanography. , 2002, Optics express.

[6] D H Burns,et al. Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens , 1993, Journal of microscopy.

[7] Masud Mansuripur,et al. Distribution of light at and near the focus of high-numerical-aperture objectives , 1986 .

[8] Ernst H. K. Stelzer,et al. Method to calculate lateral and axial gain factors of optical setups with a large solid angle , 1999 .

[9] E. Stelzer,et al. Optical trapping of dielectric particles in arbitrary fields. , 2001, Journal of the Optical Society of America. A, Optics, image science, and vision.