A High Resolution CCD Intravital Fluorescent Microscope for Observation of Coronary Smaller Microvessels in a Beating Heart

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We developed a needle-probe intravital microscope with CCD (charge coupled device), by which we can observe coronary microvessels (D>50 mum) in a beating heart with and without fluorescent dye (Circ.Res., 72 : 939-946,1993, Circ.Res.75 : 393-399,1994). To analyze smaller pre- and post-capillary microvessels, we developed a new high resolution CCD system, which consists of a needle probe, a camera body, a lens, a light guide and source, a high definition monitor and a high definition videocassette recorder. The camera has a high resolution CCD image sensor with 1940 x 1040 pixels. The needle-probe contains a relay lens (diameter : 4 mm ; length : 140 mm) to increase resolution. Images focused on the CCD through a relay lens are converted to high definition video signals every 33 ms. To examine the accuracy of this system, a micro scale pattern was observed. The spatial resolution was about 2.5 mum for the x200objective. We could clearly observe small pre- and post-capillary microvessels in subepicardium in a beating heart, which was difficult by our preview system. We also apllied our system to visualize renal microcirculation and found that the images of afferent and efferent arterioles were clealy observed by our system. In conclusion, our new high resolution needle-probe CCD intravital microscope is a powerful tool to observe small pre- and post-capillary microvessels (D<50 mum) in a beating heart.