1. How do I use my Fluor-Ref Slides?
As with any other reference or calibration slide, replace your sample with a Fluor-Ref slide. While looking directly at the slide, bring the objective close to the surface (nearly touching) then, looking down the eyepiece, focus away from the slide.
You can find the surface easily. Typically, it will have a bit of dust and will be the brightest image plane. Focus back toward the slide slightly to acquire an image from below the surface.
2. How do I clean my Fluor-Ref slides?
Simply wipe the slide down with alcohol. MME recommends either wetting a piece of lens tissue or using an alcohol wipe. Other materials, such as KimWipe, may scratch the surface.
3. How resistant are Fluor-Ref slides to immersion oil?
Our tests indicate that the slides will soften if you leave immersion oil on the slide for extended periods of time (ex: a week or more). For general maintenance, we suggest wiping the slide down with alcohol after using immersion oil.
4. How stable are Fluor-Ref slides?
Dr. P. C. Cheng of SUNY Buffalo, inventor of these slides, subjected them to full intensity UV excitation for over an hour and found a very small amount of photobleaching, just at the surface. Based on these tests, the slides are not expected to show any significant amount of photobleaching under normal use.
5. Can I use Fluor-Ref slides to acquire a system response for background correction?
Absolutely. Just image the Fluor-Ref slide as describe in #1, above. Use the image from below the surface as the system response and follow the procedure outlined in your image analysis program for background correction.
6. Can I use Fluor-Ref slides with faint fluorescence?
If you are working with faint fluorescence, you may want to use some neutral density filtration or reduce the intensity using functions in your image analysis software