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Journal of Biomedical Optics • Open Access

Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging
Author(s): Qiaole Zhao; Ben Schelen; Raymond Schouten; Rein van den Oever; Rene Leenen; Harry van Kuijk; Inge M. Peters; Frank Polderdijk; Jan T. Bosiers; Marcel Raspe; Kees Jalink; Jan Geert Sander de Jong; Bert van Geest; Karel Stoop; Ian T. Young

Paper Abstract

We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.

Paper Details

Date Published: 18 December 2012
PDF: 14 pages
J. Biomed. Opt. 17(12) 126020 doi: 10.1117/1.JBO.17.12.126020
Published in: Journal of Biomedical Optics Volume 17, Issue 12
Show Author Affiliations
Qiaole Zhao, Technische Univ. Delft (Netherlands)
Ben Schelen, Technische Univ. Delft (Netherlands)
Raymond Schouten, Technische Univ. Delft (Netherlands)
Rein van den Oever, Technische Univ. Delft (Netherlands)
Rene Leenen, Teledyne DALSA (Netherlands)
Harry van Kuijk, Teledyne DALSA (Netherlands)
Inge M. Peters, Teledyne DALSA (Netherlands)
Frank Polderdijk, Teledyne DALSA (Netherlands)
Jan T. Bosiers, Teledyne DALSA (Netherlands)
Marcel Raspe, The Netherlands Cancer Institute (Netherlands)
Kees Jalink, The Netherlands Cancer Institute (Netherlands)
Jan Geert Sander de Jong, Lambert Instruments BV (Netherlands)
Bert van Geest, Lambert Instruments BV (Netherlands)
Karel Stoop, Lambert Instruments BV (Netherlands)
Ian T. Young, Technische Univ. Delft (Netherlands)

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