Abstract
Cellular states are strongly influenced by cell-cell and cell-matrix interactions. Traditional cytometry has limited ability to quantify structure and biochemistry of intact tissues in three dimensions (3D). We extend the technique of image cytometry to quantify 3D tissue physiology and pathology states with subcellular resolution. Based on a high-speed multiphoton microscope for 3D imaging and an automated computercontrolled specimen stage, we can quantify cellular and tissue morphology and biochemistry in a high throughput manner. With the excellent penetration depth of multiphoton microscopy, it can assay tissue structures with subcellular resolution down to a few hundred micrometers in most tissues. The throughput of this instrument can be best quantified by the rate that tissue volume can be assayed. For typical tissue specimen, we can assay a tissue volume of 3 mm × 3 mm × 100 µm (width, height and depth respectively) with 1 µm3 resolution within 2 hours. If the tissue volume contains tightly packed, stratified cellular layers, this instrument samples up to approximate 200 cells per second.
© 2005 Optical Society of America
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