Proceedings of SCANNING 2006 April 25-27, 2006 Washington, D.C., USA: Tuesday, April 25

摘要

The light microscope is arguably the most ubiquitous piece of equipment in the modern biological or physical science research laboratory. It has applications from archaeology to metrology to zoology. Imaging of fluorescent samples, especially fluorescently labeled biological samples, is one of the more powerful applications of this instrumentation. A significant limitation is that images produced in conventional transmitted light and epi-fluorescent modes are essentially 2-dimensional projections of a three dimensional volume, be it a section, whole tissue or a cell culture. In addition, and especially for thicker, light scattering samples, structures outside of the plane of focus can contribute noise to the image causing blurring. To acquire high resolution volume information from a sample with a conventional light microscope it is necessary for the specimen to be fixed (in the case of biological samples,) sectioned, stained, imaged, digitized and reconstructed. This is a tedious procedure which can introduce artifacts at every step. The introduction of the Laser Scanning Confocal Microscope revolutionized 3-dimensional imaging with light microscopy. It forms images ("optical sections") with an inherently narrow depth-of-field by detecting light only from the plane of focus. By using motorized focus controls, multiple sections can be acquired at regular intervals to build up a volume data set.