The attachment of mineralised tissues to coverslips for observing dynamic events by confocal microscopy.

Abstract

In confocal scanning optical fluorescence microscopy, using high-aperture oil immersion lenses, the best images are obtained for focus planes immediately under the coverslip, or under a continuation of the coverslip with a medium with the same, high refractive index. Therefore placing the coverslip with a layer of oil underneath it is an advantage if the sample will allow it. If experimental manipulations that can displace the coverslip are involved,--for example, attempts to make dynamic observation of fluid flow--the coverslip is best cemented to the sample with a high-index material. This report describes the use of adhesive systems developed for restorative dentistry to achieve a durable attachment of dental tissues to microscope coverslips. The technique described in this paper has been used for monitoring real-time fluid movement in dentine. The samples were examined with a high-frame-rate confocal microscope (a tandem scanning microscope). The adhesive technology also could be utilised in the microscopic preparation of other porous translucent materials.