Photon budget analysis for fluorescence lifetime imaging microscopy

We have constructed a mathematical model to analyze the photon efficiency of frequency-domain fluorescence lifetime imaging microscopy (FLIM). The power of the light source needed for illumination in a FLIM system and the signal-to-noise ratio of the detector have led us to a photon “budget.” These measures are relevant to many fluorescence...

Photonic calibration for fluorescence microscopy

Based upon a collection of compact LEDs (light-emitting diodes) and a compact photodiode, we have developed a calibration tool for fluorescence microscopes that are used as digital imaging devices. The entire device (excluding a USB connector) measures 25 mm x 80 mm x 12 mm. Virtually all commonly-used fluorophores can be simulated with one of...

Absolute fluorescence calibration

While fluorescence microscope systems remains an essential tool in modern biology and medical work, no compact instrumentation has been developed for the rapid calibration of such systems. Almost invariably results are presented in terms of the [AU], "arbitrary units". To remedy this situation we have developed a small, portable instrument - the...

Three-dimensional analysis tool for segmenting and measuring the structure of telomeres in mammalian nuclei

Quantitative analysis in combination with fluorescence microscopy calls for innovative digital image measurement tools. We have developed a three-dimensional tool for segmenting and analyzing FISH stained telomeres in interphase nuclei. After deconvolution of the images, we segment the individual telomeres and measure a distribution parameter we...

LEDs for fluorescence microscopy

Traditional light sources for fluorescence microscopy have been mercury lamps, xenon lamps, and lasers. These sources have been essential in the development of fluorescence microscopy but each can have serious disadvantages: lack of near monochromaticity, heat generation, cost, lifetime of the light source, and possible distortions due to...