To make a calibration slide:

  1. Dissolve the dye in a solution with a refractive index that is similar to your specimen. The dye solution should be completely dissolved and should contain no aggregates.

  2. Place 4 microliters of the appropriate dye solution on the center of a coverslip.

  3. Pick up the drop (and coverslip) with a clean slide.

  4. Quickly invert the slide and seal the coverslip before the dye solution spreads to the edges. Be careful to not contaminate the microscope objectives with dye.

Note:  Calibration tables are generated from a uniformly fluorescent slide. You can use the fluorescent plastic calibration slides available from GE Healthcare or make a calibration slide of your own.