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Research Papers

Dynamic autofocus for continuous-scanning time-delay-and-integration image acquisition in automated microscopy

[+] Author Affiliations
Miguel E. Bravo-Zanoguera

University of California, San Diego, Department of Bioengineering, La Jolla, California 92093 and Autonomous University of Baja California, Faculty of Engineering, Mexicali, Baja California, 21280 México

Casey A. Laris

University of California, San Diego, Department of Bioengineering, La Jolla, California 92093 and Dipath Inc., Solana Beach, California 92075

Lam K. Nguyen

University of California, San Diego Department of Bioengineering, La Jolla, California 92093 and Burnham Institute, 10901 North Torrey Pines Road, La Jolla, California 92037

Mike Oliva

University of California, San Diego, Department of Bioengineering, La Jolla, California 92093 and Jtec Surgical, Inc., San Diego, California 92107

Jeffrey H. Price

Burnham Institute for Medical Research, 10901 North Torrey Pines Road, La Jolla, California 92037 and University of California, San Diego, Department of Bioengineering, La Jolla, California 92093 and Vala Sciences, Inc., San Diego, California 92121

J. Biomed. Opt. 12(3), 034011 (June 11, 2007). doi:10.1117/1.2743078
History: Received February 15, 2006; Revised December 28, 2006; Accepted February 27, 2007; Published June 11, 2007; June 28, 2007
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Efficient image cytometry of a conventional microscope slide means rapid acquisition and analysis of 20 gigapixels of image data (at 0.3-μm sampling). The voluminous data motivate increased acquisition speed to enable many biomedical applications. Continuous-motion time-delay-and-integrate (TDI) scanning has the potential to speed image acquisition while retaining sensitivity, but the challenge of implementing high-resolution autofocus operating simultaneously with acquisition has limited its adoption. We develop a dynamic autofocus system for this need using: 1. a “volume camera,” consisting of nine fiber optic imaging conduits to charge-coupled device (CCD) sensors, that acquires images in parallel from different focal planes, 2. an array of mixed analog-digital processing circuits that measure the high spatial frequencies of the multiple image streams to create focus indices, and 3. a software system that reads and analyzes the focus data streams and calculates best focus for closed feedback loop control. Our system updates autofocus at 56Hz (or once every 21μm of stage travel) to collect sharply focused images sampled at 0.3×0.3μm2/pixel at a stage speed of 2.3mms. The system, tested by focusing in phase contrast and imaging long fluorescence strips, achieves high-performance closed-loop image-content-based autofocus in continuous scanning for the first time.

Figures in this Article
© 2007 Society of Photo-Optical Instrumentation Engineers

Citation

Miguel E. Bravo-Zanoguera ; Casey A. Laris ; Lam K. Nguyen ; Mike Oliva and Jeffrey H. Price
"Dynamic autofocus for continuous-scanning time-delay-and-integration image acquisition in automated microscopy", J. Biomed. Opt. 12(3), 034011 (June 11, 2007). ; http://dx.doi.org/10.1117/1.2743078


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