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The samples were cast in mini-cubes of polyester resin (Excel Technologies, Inc., Enfield, CT). The resin was allowed to cure for 24 hours at room temperature and under ambient light. The cubes were then ground to expose the cross-sections, and dry polished with 400 and 600 grit wet-dry papers and Micro-Mesh polishing cloths, with grits from 1500 to 12,000.
The cast samples were analyzed and photographed using a Nikon Eclipse 80i epifluorescence microscope equipped with an EXFO X-Cite 120 Fluorescence Illumination System fiberoptic halogen light source and a polarizing light base using SPOT Advanced software (v. 4.6) for digital image capture and Adobe Photoshop CS for digital image management. The samples were photographed in reflected visible and ultraviolet light using a UV-2A filter with 330-380 nm excitation, 400 nm dichroic mirror and a 420 nm barrier filter and a BV-2A filter with 400-450 nm excitation, 455 nm dichroic mirror and a 470 nm barrier filter. Photographs were taken at 200X and 400X magnifications.
The samples were also stained with three fluorescent stains to characterize the binding media in the various layers and to provide a better comparison between the different materials present in the layers.
The following fluorescent stains were used for examination of the samples:  
  • Triphenyl tetrazolium chloride (TTC) 4.0% in ethanol to identify the presence of carbohydrates (starches, gums, sugars). Positive reaction color is dark red or brown.
  • 2, 7 Dichlorofluorescein (DCF) 0.2% in ethanol to identify the presence of saturated and unsaturated lipids (oils). Positive reaction for saturated lipids is pink and unsaturated lipids is yellow.
  • Rhodamine B (RHOB) 0.06% in ethanol to identify the presence of oils. Positive reaction color is bright orange.

Information Provided by Ultraviolet Light Microscopy

When viewed under visible light, cross-sections which contain ground, paint and varnish may often be difficult to interpret, particularly because clear finish layers look uniformly brown or tan. It may be impossible using only visible light to distinguish between multiple varnish layers. Illumination with ultraviolet light provides considerably more information about the layers present in a sample because different organic, and some inorganic, materials autofluoresce (or glow) with characteristic colors.  
There are certain fluorescence colors which indicate the presence of specific types of materials. For example: shellac fluoresces orange (or yellow-orange) when exposed to ultraviolet light, while plant resin varnishes (typically amber, copal, sandarac and mastic) fluoresce bright white. Wax does not usually fluoresce; in fact, in the ultraviolet it tends to appear almost the same color as the polyester casting resin. In visible light wax appears as a somewhat translucent white layer. Paints and glaze layers which contain resins as part of the binding medium will also fluoresce under ultraviolet light at high magnifications. Other materials such as lead white, titanium white and hide glue also have a whitish autofluorescence.  
There are other indicators which show that a surface has aged, such as cracks which extend through finish layers, accumulations of dirt between layers, and sometimes diminished fluorescence intensity, especially along the top edge of a surface which has been exposed to light and air for a long period of time.