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Arichard:Protocols/Microscope bulb change
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=== Alignment === * Find a sample that will fluoresce evenly. ** Cardstock works well (cellulose is autofluorescent). * The sample illumination comes from two sources: ** The bulb. ** The mirror image from the back of the bulb. * Alignment involves putting these two sources (bright dots) on top of each other as much as possible. ** They cannot align perfectly, but you can get close. * There are three knobs used for alignment: ** The collector focus knob, located on the cylindrical housing. ** The horizontal alignment knob, located on the bulb housing, parallel to the bench. ** The vertical alignment knob, located on the bulb housing, diagonal to the bench. * Alignment should be checked periodically. ==== Protocol ==== # Power up, select 4X objective and GFP. Position sample under illumination. # Adjust the collector focus until the two illumination sources are resolved as much as possible. # Adjust vertical and horizontal alignment knobs until the two sources are on top of each other as much as possible. # Back off on the collector focus to spread the illumination over the whole sample. ## The sample should fluoresce evenly if alignment was done properly. # Remember to burn in the bulb, 2-4 hours. Overnight is also OK.
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