Work

Optimizing the performance of dual-axis confocal microscopes via Monte-Carlo scattering simulations and diffraction theory

Year: 2013

Type: article

Abstract: Dual-axis confocal (DAC) microscopy has been found to exhibit superior rejection of out-of-focus and multiply scattered background light compared to conventional single-axis confocal microscopy. DAC microscopes rely on the use of separated illumination and collection beam paths that focus and intersect at a single focal volume (voxel) within tissue. While it is generally recognized that the resolution and contrast of a DAC microscope depends on both the crossing angle of the DAC beams, 2θ, and the focusing numerical aperture of the individual beams, α, a detailed study to investigate these dependencies has not been performed. Contrast and resolution are considered as two main criteria to assess the performance of a point-scanned DAC microscope (DAC-PS) and a line-scanned DAC microscope (DAC-LS) as a function of θ and α. The contrast and resolution of these designs are evaluated by Monte-Carlo scattering simulations and diffraction theory calculations, respectively. These results can be used for guiding the optimal designs of DAC-PS and DAC-LS microscopes.

Source: Journal of Biomedical Optics

DOI: 10.1117/1.jbo.18.6.066006

Authors: Ye Chen, Jonathan T. Liu

Institution: Stony Brook University

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $1,675

Funders: Stony Brook University, National Institute of Biomedical Imaging and Bioengineering, National Institute of Dental and Craniofacial Research