Further developments in addressing depth-variant 3D fluorescence microscopy imaging

Sreya Ghosh; Lutz Schaefer; Dietwald Schuster; Chrysanthe Preza

doi:10.1117/12.2045856

12 March 2014 Further developments in addressing depth-variant 3D fluorescence microscopy imaging

Sreya Ghosh, Lutz Schaefer, Dietwald Schuster, Chrysanthe Preza

Proceedings Volume 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI; 89490Q (2014) https://doi.org/10.1117/12.2045856
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

Three-dimensional (3D) imaging with optical sectioning microscopy uses computational methods to obtain the true fluorescence distribution by ameliorating the effect of defocus, spherical aberration and noise. Inverse algorithms improve image quality at a fraction of the cost of implementing an optical system by accurate modeling of the imaging system. Good inverse imaging algorithms need to be accurate as well as fast. Better understanding of the image formation model is vital to obtain improved restoration through model-based algorithms. Forward imaging models based on a depth-varying point-spread function (DV-PSF) leads to a substantial improvement in the resulting images because it accounts for depth-induced aberrations present in the imaging system. PSFs at every layer can be represented using their principal components. Computation of the forward imaging model using a principal component analysis (PCA) representation of the DV-PSF requires fewer convolutions than a strata based approach investigated in the past. In this paper we present a new algorithm for maximum likelihood image restoration developed based on a PCA representation of the DV-PSF and an accelerated conjugate gradient (CG) iteration scheme. Results obtained with this PCA-CG algorithm from both simulated and experimental fluorescence microscope data are discussed and compared with results obtained from a CG iteration method based on the strata model and linear interpolation of the DV-PSF. The performance of the PCA-CG algorithms is shown to be promising for practical applications.

Citation Download Citation

Sreya Ghosh, Lutz Schaefer, Dietwald Schuster, and Chrysanthe Preza "Further developments in addressing depth-variant 3D fluorescence microscopy imaging", Proc. SPIE 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI, 89490Q (12 March 2014); https://doi.org/10.1117/12.2045856

ACCESS THE FULL ARTICLE

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