Multi-colour TIRF

STFC has developed a fluorescence measurement technique that provides simultaneous profiling of the sample surface thereby increasing accuracy of fluorescence measurements and advanced real time monitoring of bimolecular activities. The technique would be an excellent addition to the capabilities of existing TIRF microscopes and STFC therefore invites partners in this field to exploit this innovative technique.





Fluorescence is the emission of electromagnetic radiation from a material following excitation of the material by electromagnetic radiation, or the impact of electron.  Fluorescence microscopy is a form of microscopy in which the specimen is irradiated at wavelengths which will excite fluorophores (or fluorochromes), present naturally or artificially introduced, within the specimen.

TIRF (Total Internal Reflection Fluorescence) microscopy is a specialised form of fluorescence microscopy, which uses an evanescent wave to selectively illuminate and excite fluorophores in a restricted region of the specimen.

STFC’s technology, through the use of two radiation beams, excites the fluorophores on the specimen (TIRF microscopy) as well as profiles the surface of the specimen (Profilometry) simultaneously. The radiation beams for excitation may also be provided in different polarization states in order to be able to measure changes in orientation of the dipoles of the fluorophores. Among its various applications, the technology through quick refreshing of captured images and the inherent profiling ability, would allow observation of conformation changes in tagged surface proteins or receptors in cell membranes in real time.



The radiation beam consists of two components – an excitation beam and a profiling beam. The latter measures the profile of a portion of the sample surface.



The provision of the two beams allows the normalization of measured fluorescence to account for irregularities in profile as well as the observation of conformational changes in proteins and other biomolecules on the surface of sample in real time.



Fluorescent Microscopy



Patent Information:
Country Serial No.
United States 12/595094
United Kingdom 08719001.3
Japan JP2010-503572
India 1857/MUMNP/2009
Germany 8719001.3
France 8719001.3
China 200880012484.4
For Information, Contact:
Elizabeth Bain
IP Manager
STFC Innovations
+44 (0) 1925 60 3680