Dye free automated cell counting and analysis

Authors

  • Dietrich Dehlinger,

    1. Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
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  • Lynn Suer,

    1. Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
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  • Maher Elsheikh,

    1. Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
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  • José Peña,

    1. Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
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  • Pejman Naraghi-Arani

    Corresponding author
    1. Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
    • Lawrence Livermore National Laboratory, P.O. Box 808 L-452, Livermore, California 94550; telephone: 925-422-5665; fax: 925-422-2282
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Abstract

We have developed an automated cell counting method that uses images obtained at multiple focal heights to enumerate cells in confluent culture. By taking the derivative of image intensity with respect to focal height using two complementary images, we are able to count high-density monolayers of cells over a large image area. Our method resists errors arising from variability in the focal plane caused by flatness or tilt non-uniformities with a minimal amount of focal plane alignment, allowing the automated collection of images across a large area. Biotechnol. Bioeng. © 2013 Wiley Periodicals, Inc.

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