CONFERENCE PROCEEDINGS
Advanced Search
Home > Proceedings > Volume 10069 > Article
Paper
21 February 2017 Enhancing resolution and contrast in second-harmonic generation microscopy using an advanced maximum likelihood estimation restoration method
Mayandi Sivaguru, Mohammad M. Kabir, Manas Ranjan Gartia, David S. C. Biggs, Barghav S. Sivaguru, Vignesh A. Sivaguru, Zachary T. Berent, Amy J. Wagoner Johnson, Glenn A. Fried, Gang Logan Liu, Sakthivel Sadayappan, Kimani C. Toussaint Jr.
Author Affiliations +
Proceedings Volume 10069, Multiphoton Microscopy in the Biomedical Sciences XVII; 1006908 (2017) https://doi.org/10.1117/12.2256534
Event: SPIE BiOS, 2017, San Francisco, California, United States
Abstract
Second-harmonic generation (SHG) microscopy is a label-free imaging technique to study collagenous materials in extracellular matrix environment with high resolution and contrast. However, like many other microscopy techniques, the actual spatial resolution achievable by SHG microscopy is reduced by out-of-focus blur and optical aberrations that degrade particularly the amplitude of the detectable higher spatial frequencies. Being a two-photon scattering process, it is challenging to define a point spread function (PSF) for the SHG imaging modality. As a result, in comparison with other two-photon imaging systems like two-photon fluorescence, it is difficult to apply any PSF-engineering techniques to enhance the experimental spatial resolution closer to the diffraction limit. Here, we present a method to improve the spatial resolution in SHG microscopy using an advanced maximum likelihood estimation (AdvMLE) algorithm to recover the otherwise degraded higher spatial frequencies in an SHG image. Through adaptation and iteration, the AdvMLE algorithm calculates an improved PSF for an SHG image and enhances the spatial resolution by decreasing the full-width-at-halfmaximum (FWHM) by ~20%. Similar results are consistently observed for biological tissues with varying SHG sources, such as gold nanoparticles and collagen in porcine feet tendons. By obtaining an experimental transverse spatial resolution of ~400 nm, we show that the AdvMLE algorithm brings the practical spatial resolution closer to the theoretical diffraction limit. Our approach is suitable for adaptation in micro-nano CT and MRI imaging, which has the potential to impact diagnosis and treatment of human diseases.
© (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation Download Citation
Mayandi Sivaguru, Mohammad M. Kabir, Manas Ranjan Gartia, David S. C. Biggs, Barghav S. Sivaguru, Vignesh A. Sivaguru, Zachary T. Berent, Amy J. Wagoner Johnson, Glenn A. Fried, Gang Logan Liu, Sakthivel Sadayappan, and Kimani C. Toussaint Jr. "Enhancing resolution and contrast in second-harmonic generation microscopy using an advanced maximum likelihood estimation restoration method", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006908 (21 February 2017); https://doi.org/10.1117/12.2256534
ACCESS THE FULL ARTICLE
ORGANIZATIONAL
Sign in with credentials provided by your organization.
INSTITUTIONAL
Select your institution to access the SPIE Digital Library.
SELECT YOUR INSTITUTION
PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.
PERSONAL SIGN IN
No SPIE Account? Create one
PURCHASE THIS CONTENT
SUBSCRIBE TO DIGITAL LIBRARY
50 downloads per 1-year subscription
Members: $195
Non-members: $335 ADD TO CART
25 downloads per 1 - year subscription
Members: $145
Non-members: $250 ADD TO CART
PURCHASE SINGLE ARTICLE
Includes PDF, HTML & Video, when available
Members: $17.00
Non-members: $21.00 ADD TO CART
PROCEEDINGS
 8 PAGES
DOWNLOAD PAPER SAVE TO MY LIBRARY
GET CITATION
Advertisement
Advertisement
RIGHTS & PERMISSIONS
Get copyright permission  Get copyright permission on Copyright Marketplace
KEYWORDS
Second-harmonic generation
Microscopy
Image processing
Point spread functions
Gold
Nanoparticles
Image resolution
RELATED CONTENT
Subscribe to Digital Library
Receive Erratum Email Alert
Back to Top