Hirotoshi Terada

Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera

We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (~0.2 μm) and temporal (~50 psec) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens including stained pollen grains and fibroblast cells expressing green fluorescent protein. The lifetime values obtained matched well with those reported earlier by other groups for these same specimens. Potential applications of the present system include the measurement of intracellular physiology and Fluorescence Resonance Energy Transfer (FRET) imaging which are discussed in the context of live cell imaging.We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (∼0.2 μm) and temporal (∼50 ps) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in the literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens including stained pollen grains and fibroblast cells expressing green florescent protein. The lifetime values obtained matched well with those reported earlier by other groups for these same specimens. Potential applications of the present system include the measurement of intracellular physiology and fluorescence resonance energy transfer imaging, which are discussed in the context of live cell imaging.

Automatic processing scheme for low laser invasiveness electro optical frequency mapping mode

Electro optical techniques are efficient backside contactless techniques usually used for design debug and defect location in modern VLSI. Unfortunately, the signal to noise ratio is quite low and depends on laser power with potential device stress due to long acquisition time or high laser power, especially in up to date technologies. Under these conditions, to maintain a good signal or image quality, specific signal or image processing techniques can be implemented. In this paper, we proposed a new spatial filtering by stationary wavelets and contrast enhancement which allows the use of low laser power and short acquisition time in image mode.

Multiphoton fluorescence lifetime imaging microscopy (FLIM) using a streak camera

We report the development, calibration and biomedical application of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (approximately 0.2 μm) and temporal (approximately 50 psec) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens in the context of quantitative measurement of fluorescence resonance energy transfer (FRET).

High spatial and temporal resolution thermal imaging for LSI circuits with phase microscopy

A new thermal imaging method that senses the change in Si index of refraction as a function of temperature to visualize temperature distributions of LSI circuits is described. The resolution is not limited by the heat radiation wavelength but by the sensing light source, which is usually around 1µm. The method further extends the application into the areas that require both high spatial and temporal resolution by utilizing a pulsed light source.