( dual-light transilluminator...)Detect Released Oligosaccharides Using the Eagle Eye II,,,Still Video Imaging System

dual-light transilluminator was used. The bands in the profiling gel (Figure 1, Panel A) were excited using the overhead 312-nm light that comes with the Eagle Eye II unit. The UV lights from the transilluminator were not used. The lens aperture was set at an f-stop of 1.2 and was equipped with Stratagenes SYBR Green band-pass filter. A negative image was created using dynamic integration at 25 frames/0.8 seconds, with incremental counts of 25 frames/0.8 seconds for 9.6 seconds.

Next, the gel was imaged using Glykos SE2000 gel imager (Figure 1, Panel B), which is similar to Bio-Rads Glyco Doc Imager. An image rectangle was used to match the outline of the gel that was imaged using the Eagle Eye II still video system. Saturation was set at 0.0%.

Conclusions

Using Stratagenes Eagle Eye II still video system, it takes less than 5 minutes to image a FACE gel and produces comparable results to those obtained with the Glyko SE2000 gel imager.

We discovered that the SYBR Green filter and the overhead 312-nm UV bulb must be attached for oligosaccharide profiling gels because the dual-light UV transilluminator (312 nm) was too intense for the gels fluorophore-labeled oligosaccharides: it becomes quickly bleached from the multiple bulbs in the UV lightbox. We also found that the gel must be imaged within 15 minutes of the gel run for optimum band sharpness; after the same amount of time at room temperature, the bands spread significantly.

While the contrast in banding patterns appears higher in the gel image taken using Glykos gel imager, this is probably due to the Glyko cameras shorter focal distance. However, the same banding numbers and definitions are seen in both gels. Additionally, Stratagenes gel analytical software program can be customized to analyze and quanti
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