Measurement steps  

Place the focused detection spot into the sample: Measurement near the glass bottom allows miniaturized sample volumes.

Start the fluctuation analysis by recording the photon count trace: Data acquisition times as short as one second are sufficient to record large datasets allowing reliable correlation analysis.

Determine the correlation: By applying the correlation function to the photon count trace the signal is scanned for self-similar patterns, giving rise to a characteristic correlation function.

Fitting to appropriate model: By defining a biophysical diffusion model the number of molecules, size of particles and other parameters can be calculated from the correlation curve.

Assessing fit quality: How well the end predictions describe the system examined can be assessed from the deviations between correlated and theoretically derived models.

We recently published with our customer Merck KGaA the use of FCCS to support the identification of Methionine Aminopeptidase-2 (MetAP-2) inhibitor M8891.

See our new publication on the quantifiaction of target occupancy in tissue:

Check out our new GPCR-ligand interaction assay!