Single MoleculeFluorescence Microscopy

Single Molecule Microscopy

Classical biological techniques focus on describing the behavior of large ensembles of molecules. However, this doesn’t allow for the identification of subpopulations within the ensemble or show how these subpopulations behave under differing experimental conditions. The only way to investigate the behavior of single molecules or subpopulations is under strict experimental conditions, guaranteeing that each molecule is in the same state, one at a time.

Single-molecule fluorescence microscopy represents a subset of fluorescence microscopy that uses fluorescent tags to detect and analyze individual single molecules. This allows the activity of single molecules to be visualized with high signal-to-noise without disturbing the physiological conditions of the biological system.

Single molecule imaging 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.

Prime 95B

Extremely sensitive, 95% quantum efficient sCMOS camera with 11 µm pixels and EMCCD level detection.

Go beyond EMCCD for single molecule imaging 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.

Single Molecule Microscopy samples
Prime BSI photo

Prime BSI

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 single molecule imaging.

The Prime BSI offers an alternative to the Prime 95B when resolution is more important than extreme sensitivity.

Single Molecule Microscopy samples