Related Experiment Video
Updated: May 15, 2026
06:01
Fluorescence Lifetime Macro Imager for Biomedical Applications
Published on: April 7, 2023
Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence
Qiaole Zhao1, Ben Schelen, Raymond Schouten
1Delft University of Technology, Lorentzweg 1, 2628 CJ, Delft, The Netherlands.
Journal of Biomedical Optics
|January 18, 2013
Summary
Researchers developed a novel all-solid-state camera for frequency-domain fluorescence lifetime imaging microscopy (FLIM). This modulated, electron-multiplied FLIM (MEM-FLIM) camera offers higher resolution and improved image quality compared to traditional systems.
Area of Science:
- Microscopy
- Biophotonics
- Imaging Technology
Background:
- Frequency-domain fluorescence lifetime imaging microscopy (FLIM) traditionally requires image intensifiers.
- Existing FLIM systems face limitations in resolution and image quality.
Purpose of the Study:
- To develop and evaluate a novel all-solid-state camera for frequency-domain FLIM.
- To eliminate the need for image intensifiers in FLIM systems.
- To assess the performance characteristics of the new camera.
Main Methods:
- Development of a pixel-level modulated, application-specific charge-coupled device camera.
- Characterization of camera parameters including noise, dark current, gain, sensitivity, and linearity.
- Lifetime measurements on various samples: fluorescein solution, fixed GFP cells, and live GFP-actin stained cells.
- Comparison with a conventional microchannel plate (MCP)-based FLIM system.
Main Results:
- The novel modulated, electron-multiplied FLIM (MEM-FLIM) camera was successfully built and evaluated.
- Experimental characterization confirmed key camera performance metrics.
- MEM-FLIM demonstrated superior resolution and image quality compared to MCP-based FLIM.
- Successful lifetime measurements were achieved on diverse biological samples.
Conclusions:
- The developed MEM-FLIM camera is a viable alternative to conventional FLIM systems.
- This technology offers enhanced resolution and image quality for frequency-domain FLIM.
- The MEM-FLIM camera presents a new avenue for advanced FLIM applications.
Related Concept Videos
01:37Super-resolution Fluorescence Microscopy
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
01:16Confocal Fluorescence Microscopy
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...