Insti-Fluor Combo2 test slides
These slides have two sample wells that have been treated to be "Insti-Spore" type test slides with all three DNA dyes. The well closest to the label is a typical Insti-Spore well using SYTO as the fluorophore. The sample well furthest from the label is treated with both DAPI and PI. Thus, both wells will reveal Bacillus endospores.
Here's how to use Combo2 test slides
Set aside the 40mm coverslip
Use the color tab to handle the coverslip.
Remove the seal top
This reveals the adhesive layer.
Add 10µl sample suspension (in water).
Add 10µl sample suspension to both sample wells.
Same sample to both wells
This will allow you to see endospores regardless of possible conflicting autofluorescence colors.
Stir
It's best to distribute the sample throughout the well.
Dry at 42C for about 5min
This gives time for the reaction and staining to occur.
Please do not subject the slide to a temperature higher than 42C.
While the slide is drying set your pipetter to 7µl
Also, find your immersion oil.
Add 7µl immersion oil to both wells
Immersion oil increases resolution and the length of time a sample will fluoresce before photobleaching.
Use the color tab to apply the 40mm coverslip.
Slide is ready to be imaged
Use Blue excitation (WB) for the well closest to the label. Use either UV (WU) or Green (WG) excitation to examine the well furthesty from the label
Bacillus endospores (E) can be seen in all three probe colors. Bacillus vegetative cells (V) are also visible.