TIRF
Total Internal Reflection Fluorescence Microscopy (TIRFm) illuminates only the portion of the sample close to the coverslip via the evanescent wave generated when the excitation laser is reflected off the sample/glass interface. The result is a very narrow band of excitation resulting in excellent image contrast. This also means that most of the excitation light is reflected back into the light path of the microscope. The emission filters must attenuate an enormous amount of laser illumination returning through a microscope objective lens, often at extreme angles-of-incidence. In some applications, such as single-molecule TIRF (including PALM and STORM imaging in TIRF mode), the ratio of fluorescence signal-to-illumination could be as low as 1 in 1015:
Three things are necessary to get the signal you need: an emission filter with superior blocking of the excitation light, an ultra-flat dichroic for distortion-free reflection of the excitation laser beam, and no-torque filter cube that can hold that dichroic stress-free.
Our complete TIRF sets remove the guesswork and provide you with the tools to obtain the highest possible signal/noise ratios by blocking the laser illumination at exceedingly high levels. Chroma’s UltraFlat dichroic mirrors mounted in our own metal TIRF cubes provide distortion-free reflection of lasers creating the perfect evanescent wave.
If you don’t see what you are looking for, please contact us. We have a large inventory of filters and a knowledgeable staff ready to help you design and build a filter to meet any specification.
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