KEYWORDS: Digital image correlation, Data modeling, Microscopy, Error analysis, Point spread functions, Data acquisition, Statistical modeling, Phase measurement, Image acquisition, 3D modeling
Knowledge of prior sample information, such as a refractive index (RI) map, can be used to improve image formation models enabling more accurate three-dimensional (3D) restoration in fluorescence microscopy. RI is an indicator of cell composition and structure that allows a more comprehensive representation of the 3D structure of a specimen than fluorescence alone. Due to the integral nature of sample phase, the challenge to compute the RI map is to decouple RI and thickness. Our work investigates the feasibility of determining RI of a specimen from differential interference contrast (DIC) microscopy data acquired by using different wavelengths in illumination. This spectral diversity in the data is exploited to determine sample thickness and RI. Results from simulated and experimental data of polystyrene bead samples are presented to analyze this approach. Phase images were estimated from the DIC data using an alternating minimization algorithm. This study shows that the maximum estimated phase delay is accurate within approximately 7 percent error relative to the 2D phase model. The sensitivity of this integrated approach allows RI to be computed within approximately 0.4 percent error relative to values from the literature.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
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.