TIRF MicroscopyFluorescence Microscopy
The resolution at which fluorescent molecules can be visualized depends greatly on the preparation of the sample and the objectives used, but this can be limited by out of focus light being collected in a focal plane.
Total internal reflection fluorescence microscopy (TIRF) makes use of specific optics to produce illumination light only at the 50-100 nm range at the interface of the slide, massively reducing out of focus light and improving the ability to detect fluorescent molecules. Because of its low light intensity and high spatial resolution, it is a key technique in live-cell imaging.
TIRF is typically a very low-light technique so one of the main challenges is to collect as many of the emitted photons as possible to maximize signal to noise ratio so a highly sensitive camera is desired.
Extremely sensitive, 95% quantum efficient sCMOS camera with 11 µm pixels and EMCCD level detection.
Go beyond EMCCD for TIRF with the back-illuminated Prime 95B, which features an equivalent level of detection but with a faster speed, larger field of view and no EM-gain aging or excess noise.
The Prime 95B allows exposure times to be lowered significantly to increase acquisition speed and reduce photobleaching and photodamage to the lowest levels possible on an sCMOS camera.
High sensitivity, 95% quantum efficient, sCMOS camera with 6.5 µm pixels and 1.0 e– read noise.
The high quantum efficiency and low read noise combined with the balanced 6.5 µm pixel size offers high sensitivity imaging whilst achieving Nyquist sampling with the most popular objective magnifications used for TIRF.
The Prime BSI offers an alternative to the Prime 95B when resolution is more important than extreme sensitivity.
Single Molecule TIRF Imaging
Cees Dekker Group
Delft University of Technology
“The Prime BSI offers several advantages such as speed, field of view and price when compared to the EMCCD. The service at Teledyne Photometrics is also very good, I was always updated, and my questions were answered quickly, Teledyne Photometrics were really willing to help out.”
Simon Watkins, Ph.D.
University of Pittsburgh
“I think the Prime 95B [Scientific CMOS camera] is a very innovative product. Partly because of the back thinning but also because the pixel size is more appropriate than conventional Scientific CMOS cameras.”
Dr. Kristina Ganzinger
Physics of Cellular Interactions Group at AMOLF
“It used to be that people would go for EMCCDs for their high quantum efficiency. However, back-illuminated sCMOS cameras now offer the same quantum efficiency but with the larger field of view which, for us, was essential for our multicolor acquisitions”