Ray, Microscope optics usually come in cubes as paired sets . . . UV excitation is usually paired with something around 470 nm as emission (okay for DAPI, Hoescht, AMCA). I would go with what Olympus suggests, being very specific about the wavelength requirements. You might need to find a fluorimeter, and scan your dyes to get a better handle on what the emission wavelengths are. If you can't find a published spectrum, this may be the only way to proceed. You might also be able to modify filter blocks . . . perhaps swap the emission filter of a set. And, yes, they're pretty expensive. Luck. MAK. >>> "ray hester" <rhester@jaguar1.usouthal.edu> 11/01/01 19:49 PM >>> Hi, Has anyone looked at filipin staining on a fluorescence microscope using standard dichroics (filipin excites around 350 and emits at wavelengths similar to FITC) - specifically, if you use the UV dichroic (for 350 excitation) will you see any fluorescence in the 515 to 545 range? I was told by an investigator that Olympus (I believe that's the manufacturer they spoke with) quoted a price of around $800 for a dichroic that would allow you to visualize filipin fluorescence, but they were hopeful that they might see at least some of that fluorescence with the UV dichroic. I haven't been able to find an emission spectrum for filipin. Also, thioflavin? I believe it excites around 430 and emits similarly to FITC. Again, I haven't seen an excitation spectrum and wonder if it would excite using the FITC dichroic. They aren't staining for filipin and thioflavin simultaneously (I hope). Any information on seeing fluorescence from these two compounds on a standard fluorescnce microscope with standard dichroics will be greatly appreciated. Thanks. Ray Hester Univ. of South Alabama rhester@jaguar1.usouthal.edu
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