UNIT 2.22 How to Build a Time-Gated Luminescence Microscope

  1. Dayong Jin1,
  2. Yiqing Lu1,
  3. Robert C. Leif2,
  4. Sean Yang2,
  5. Megha Rajendran3,
  6. Lawrence W. Miller3

Published Online: 2 JAN 2014

DOI: 10.1002/0471142956.cy0222s67

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Jin, D., Lu, Y., Leif, R. C., Yang, S., Rajendran, M. and Miller, L. W. 2014. How to Build a Time-Gated Luminescence Microscope. Current Protocols in Cytometry. 67:2.22:2.22.1–2.22.36.

Author Information

  1. 1

    Advanced Cytometry Laboratories, MQ BioFocus Research Centre & Photonics Research Centre, Macquarie University, New South Wales, Australia

  2. 2

    Newport Instruments, San Diego, California

  3. 3

    Department of Chemistry, University of Illinois at Chicago, Chicago, Illinois

Publication History

  1. Published Online: 2 JAN 2014


The sensitivity of filter-based fluorescence microscopy techniques is limited by autofluorescence background. Time-gated detection is a practical way to suppress autofluorescence, enabling higher contrast and improved sensitivity. In the past few years, three groups of authors have demonstrated independent approaches to build robust versions of time-gated luminescence microscopes. Three detailed, step-by-step protocols are provided here for modifying standard fluorescent microscopes to permit imaging time-gated luminescence. Curr. Protoc. Cytom. 67:2.22.1-2.22.36. © 2014 by John Wiley & Sons, Inc.


  • lanthanide;
  • time-gated luminescence;
  • microscopy;
  • autofluorescence