Using out-of-focus light to improve image acquisition time in confocal microscopy

Brynmor J. Davis; William C. Karl; Bennett B. Goldberg; Anna K. Swan; M. Selim Unlu

doi:10.1117/12.590640

24 March 2005 Using out-of-focus light to improve image acquisition time in confocal microscopy

Brynmor J. Davis, William C. Karl, Bennett B. Goldberg, Anna K. Swan, M. Selim Unlu

Proceedings Volume 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII; (2005) https://doi.org/10.1117/12.590640
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

Light that would typically be discarded at a confocal microscope's detector pinhole will be collected and processed to allow a reduced spatial sampling rate and thus an improved image acquisition time. It is shown that collecting and appropriately processing the out-of-focus light will allow an axial sampling rate below that specified by the Nyquist criterion. To achieve this, a central detector pinhole and a number of out-of-focus regions are collected concurrently. This corresponds to imaging through several different channels, with differing point spread functions, in parallel. Since the spatial sampling rate is below the Nyquist frequency, aliasing occurs in the data from each of the channels. However, since the point spread functions are different, the aliasing effects are different in each channel. This allows the ensemble of aliased images to be processed into a single dealiased and deconvolved image. This potential utility of out-of-focus light is demonstrated through simulated examples for differing collection schemes and scanning rates. Results are shown for under-sampling by up to a factor of four. Collecting the out-of-focus light also improves instrument collection efficiency.

Citation Download Citation

Brynmor J. Davis, William C. Karl, Bennett B. Goldberg, Anna K. Swan, and M. Selim Unlu "Using out-of-focus light to improve image acquisition time in confocal microscopy", Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005); https://doi.org/10.1117/12.590640

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