Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging

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

doi:10.1117/1.JBO.17.12.126020

18 December 2012 Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging

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

Author Affiliations +

Journal of Biomedical Optics, Vol. 17, Issue 12, 126020 (December 2012). https://doi.org/10.1117/1.JBO.17.12.126020

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.

Citation Download Citation

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, and Ian T. Young "Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging," Journal of Biomedical Optics 17(12), 126020 (18 December 2012). https://doi.org/10.1117/1.JBO.17.12.126020

Published: 18 December 2012

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
14 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY