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Image Analysis Program for Education Design and construction of a confocal microscope for teaching and training Peng Xi, Bartelomiej Rajwa, Jim Jones, and J. Paul Robinson The confocal scanning laser microscope (CSLM) has shown its superior
abilities in lateral and axial resolution[1-3] and has been employed as a
routine tool for various microscopy purposes. However, commercially
available confocal systems are usually too expensive for most individual
researchers. Moreover, the commercial confocal system are expensive and very
difficult for users to modify it based on their own research applications [4]. For these reasons, Parker et al. built up their own
confocal setup for the measurement of Ca2+ concentration at the cellular
level [5,6]. The setup demonstrates its ability in real-time imaging of the
fluorescence of Xenopus oocytes. However, the fast imaging requirement makes
the setup complicated, and further image correction is a must. This causes
unnecessary system complication in the electronic controlling and image
reconstruction. In this work, a simplified small CSLM is constructed for diffused reflection
imaging and 3-D projection. Using the optical dissection ability of the confocal
setup, depth-resolved images are collected from a fern spore specimen, and a 3-D
reconstruction model is demonstrated. This system shows great capability in
depth-resolved scanning and 3-D reconstruction comparing with commercial CSLM.
It simplifies the confocal setup by requiring only essential confocal parts,
thus greatly reducing the difficulties for researchers building their own. As
the system is cost-efficient, easily aligned and robust, it can be applied in
experimental education for students. Suggestions are also given for modification
considerations. Therefore, it should be highly valuable for researchers
interested in having a CSLM with great flexibility on a modest budget, as well
as for educators demonstrating the principles of
confocal microscopy. Follow the navigation links to see how to achieve this -- build it and enjoy! © 2006 Purdue University Cytometry Laboratories. |
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