Chlorophyll fluorescence imaging by FLUORCAM-in allows you to present results in the form of an image, but also in the form of numbers. Analysing the data they are often divergent. Moreover, very often the measurement error appears "overflow pixel" - as it avoided lest they destroy plants?
Hi Katarzyna,
the pixel overflow usually means that you exceeded the detector sensitivity. Normally the plants are not destroyed by saturating flashes (except some very sensitive photosynthesis mutants - maybe). In order to get good results and reliable data, the sensitivity of the camera and the measuring light intensity (not the actinic light!) should be adjusted in a right way before the actual experiment takes place. A good start is to reach 10-15% of max. detector value with F0 measurement - so you end up with ~75% of max. detector limit with a saturating flash.
Make sure that your measuring light does not induce too much photosynthesis ( in an ideal world by switching on the measuring light the actual fluorescence yield shouldn't change).
I hope I could help you. If you have some more questions don't hesitate to contact me.
otherwise have fun!
Best wishes
Lars
If you mean what optics are used, their web site indicates a 7-filter wheel is an option, so you either collect all light to a bare grayscale CCD camera, or pinch of a band of color to send to the camera, for GFP v. chlorophyll for instance. I don't work with this instrument but it looks like "spectral" could refer to the color of illumination and/or the color of detection. You can flash certain colors with certain LED panels, and detect everything or just one color at a time through the filter wheel.
Overflow pixel usually means saturation. Try to lover the exposure time of your camera/detector.
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