Bio-Rad Components for Older Model Spot Cutter - Manual (Page 15)

Section 4
Fluorescent Operation in Basic Excision Tool
The PDQuest version 6.2.1 is very similar to version 6.2.0 in operating the spot cutter. There are
some new function buttons and workflow items based on using the fluorescent enclosure. For a
more complete description of the general function of the spot cutter in Basic Excision Tool mode,
see the ProteomeWorks Spot Cutter manual. The following sections describe the workflow of the
Fluorescent Enclosure items that differ from the white light operation in Basic Excision Tool.
4.1 Calibration and the UV Light Option
The software records the filter set-up with a confirmation button in the calibration. If the "Filter is
not Installed" button is clicked, because the filter has not been installed, then it indicates to the
software that the fluorescent enclosure is not being used. In this case, the UV light option in the
software will be grayed out and is not available. To activate the UV lights, the filter should be
added to the lens and the "Filter is installed" message is clicked at the beginning of the
calibration. This provides a record of how the installation was performed and the hardware that
was in place during calibration.
4.2 Acquire Fluorescent Image in Basic Excision Tool
To acquire an image in the Basic Excision Tool screen, select the “UV on (Fluor)” lights setting,
and manually enter an exposure time in seconds. Typical exposure times for SYPRO Ruby stained
2D gels are from 1 to 40 seconds. Start with short exposure times and increase the time as
needed for optimum imaging.
Fig. 18. Example PDQuest version 6.2.1 Basic Excision Tool software with SYPRO Ruby stained 2-D gel.
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