Characterization of optical-aberration-induced lateral and axial image inhomogeneity in multiphoton microscopy
Journal
Journal of Biomedical Optics
Journal Volume
13
Journal Issue
4
Date Issued
2008
Author(s)
Advisor
WOS:000259901000047
Abstract
The effects of off-axis optical aberration in multiphoton microscopy and the resulting lateral and axial image inhomogeneity are investigated. The lateral inhomogeneity of the scanning field is demonstrated by second harmonic generation (SHG) imaging of fasciae and two-photon fluorescence (TPF) microscopy of thin fluorescent samples. Furthermore, refractive index mismatch-caused intensity attenuation of the TPF signal at central and peripheral regions of the scanning frame is measured using homogeneous 10-μM sulforhodamine B samples with refractive indexes of 1.33 and around 1.465. In addition to characterizing image field convexity, we also found that image resolution degrades away from the optical axis. These effects need to be accounted for in both qualitative and quantitative multphoton imaging applications. © 2008 Society of Photo-Optical Instrumentation Engineers.
Subjects
fluorescence; multiphoton; point spread function; second harmonic generation; spherical aberration
Other Subjects
Image fields; Imaging applications; Inhomogeneities; Lateral inhomogeneity; Multi-photon microscopy; Multiphotons; Off-axis; Optical aberration; Optical axis; Point-Spread function; Spherical aberrations; Sulforhodamine B; Two photon fluorescence; Aberrations; Fluorescence; Image resolution; Nonlinear optics; Optical transfer function; Refractive index; Harmonic generation; animal; anisotropy; article; artifact; chicken; computer assisted diagnosis; cytology; evaluation; fascia; image enhancement; methodology; multiphoton microscopy; reproducibility; sensitivity and specificity; Animals; Anisotropy; Artifacts; Chickens; Fascia; Image Enhancement; Image Interpretation, Computer-Assisted; Microscopy, Fluorescence, Multiphoton; Reproducibility of Results; Sensitivity and Specificity
Type
journal article