Hi, I'm trying to determine if my protein spherulites are positive or negative by polarized light microscopy. This involves the use of a 530 or 550nm retardation plate (RP) inserted before the analyzer of a cross-polarization setup (i.e from the bottom: polarizer>sample>RP>analyzer).
Analyzer and polarizer have to be perpendicular each other, let's say one north-south, the other east-west, while the retardation plate has to be set at 45°, so either direction southwest-northeast or southeast-northwest.
In my microscope (Olympus BX61 equipped with a U-TP530 at 45°) the fast axis of the RP is southwest-northeast while in most references I found this is either not specified or the opposite (southeast-northwest).
So my question is, since I have an "opposite" set up from the ones in the reference, is also the coloring of my spherulite opposite (i.e. if positive, radial spherulites observed with a retardation plate oriented southeast-northwest show their northeast quadrant blue, should my spherulites, assuming they are also positive, show the same quadrant yellow instead)?
References:
Article The formation of spherulites by amyloid fibrils of bovine insulin
http://dx.doi.org/10.1016/j.polymer.2013.11.038
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