Overview of CircuSoft PFRET

Fluorescence resonance energy transfer (FRET) microscopy is an ideal technique for detecting proteins in their natural states in cells and tissues. Various advanced FRET microscopy techniques (wide-field, Confocal, and Multiphoton) exist for localizing the individual proteins, but they all suffer from various drawbacks including autofluorescence, detector noise, optical noise, photobleaching and spectral bleed-through. CircuSoft PFRET corrects FRET images and performs distance and efficiency analyses to give you quantitative FRET results.

Spectral bleed-through (SBT) is the single largest complication to FRET experiments, an effect due to contributions of donor and acceptor fluorescence emission into the FRET channel. SBT makes the observed FRET signal greater than the real signal. Researchers at the W. M. Keck Center for Cellular Imaging at the University of Virginia have developed a new algorithm to remove SBT and correct for the variations in fluorophore expression levels (FEL) for all intensity-based FRET techniques [references]. CircuSoft PFRET incorporates this algorithm in a user-friendly software package that includes functions for calculation of energy transfer efficiency and the distance between donor and acceptor molecules.