TIRF & TRF89900 Series
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.
We offer two versions of each our TRF89900 dual band TIRF filter sets, each providing greater levels of blocking of the TIRF lasers than our 89900 series filter sets. One version (“-EM”) utilizes additional, single band emission filters which match the quad band emission filter in the filter cube. This option requires an emission filter wheel to house the additional emission filters. The other option relies on one quad band emission filter which provides extra blocking of the TIRF laser, and is housed in the filter cube. The highest signal/noise levels are obtained with the “-EM” versions, however for workers who need very fast switching or who don’t have a filter wheel, the alternate version is an excellent option. The Ultra-Flat Dichroics in our TIRF sets are 2mm thick for distortion-free reflection of TIRF lasers with surface flatness =/> 0.5 waves/inch P-V, RWD. All catalog TIRF sets are mounted in Chroma’s metal TIRF cubes. For more information, please see the supporting materials on our TIRF filter set webpages.
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