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In recent years, the total internal reflection fluorescence microscopy (TIRFM) has become popular due to the fact the TIRFM has an ability of high sensitive fluorescence observation, especially for single moleculeimaging and analysis for biological functions around cell membrane such as exocytosis. However, optical alignment of incident laserhad been so complicated until now that we could notset up illumination for total internal reflection correctly in some cases. This time, we developed an equipment for TIRFM with easy handling. Ultra high NA objective lenses (Apo 100X NA 1.65 and Plan Apo 60X NA 1.45) enable high contrast and bright images of fluorescence because of high numerical aperture over 1.4. In TIRFM illuminator, the laser and illuminator are connected with the fiber, so almost no additional optical alignment is necessary.

Ultra High Numerical Aperture Objective Lenses and Optical System Improved Objective Type Total Internal Reflection Fluorescence Microscopy.