Practical Aspects of Fluorescence Filter Combinations
Section Overview:
Practical Aspects of Fluorescence Filter Combinations
Brief Overview of Fluorescence Filters
A wide spectrum of filter cubes is available from most major manufacturers, which now produce filter sets capable of imaging most of the common chromophores in use today.
Basic Aspects of Light Filters
Filters work by absorbing light, while others reflect unwanted light, but pass a selected region of wavelengths. Some filters operate by interference and can be adjusted to pass narrow bands of light while reflecting all others.
Introduction to Fluorophores
Laser scanning confocal microscopy relies on secondary fluorescence emission as an imaging mode, due to the high degree of sensitivity afforded by the technique coupled with the ability to specifically target structural components.
Acousto-Optic Tunable Filters (AOTFs)
Several benefits of the AOTF combine to greatly enhance the versatility of the latest generation of confocal instruments, and these devices are becoming popular for control of excitation wavelength ranges and intensity.
Interference Filters for Fluorescence Microscopy
The utilization of highly specialized thin filminterference filtershave enhanced the versatility and scope of fluorescence techniques, far beyond the capabilities afforded by the earlier use of gelatin and glass filters.
Spectral Bleed-Through Artifacts in Confocal Microscopy
Spectral bleed-through of fluorescence emission, which occurs due to the very broad bandwidths and asymmetrical spectral profiles exhibited by many of the fluorophores, is a problem that must be addressed in microscopy.