title: Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging.
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.
authors: Delft University of Technology, Lorentzweg 1, 2628 CJ, Delft, The Netherlands.
authors: Qiaole
authors: Q
authors: Zhao
authors: Ben
authors: B
authors: Schelen
authors: Raymond
authors: R
authors: Schouten
authors: Rein
authors: R
authors: van den Oever
authors: René
authors: R
authors: Leenen
authors: Harry
authors: H
authors: van Kuijk
authors: Inge
authors: I
authors: Peters
authors: Frank
authors: F
authors: Polderdijk
authors: Jan
authors: J
authors: Bosiers
authors: Marcel
authors: M
authors: Raspe
authors: Kees
authors: K
authors: Jalink
authors: Jan
authors: J
authors: Geert Sander de Jong
authors: Bert
authors: B
authors: van Geest
authors: Karel
authors: K
authors: Stoop
authors: Ian Ted
authors: IT
authors: Young
mesh_terms: D000669:Amplifiers, Electronic; D004583:Electrons; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012666:Semiconductors; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation*