Probing protein association, or oligomerization, in cells is an important problem in life sciences. Determining the size of their oligomers, or the timing of the oligomerization onset pose substantial experimental challenges, with different studies often producing contradicting results. Existing technologies either are laborious and slow or lack the bandwidth needed to discriminate between different oligomeric sizes and expression levels (i.e., concentrations).
A molecular brightness-based determination algorithm is incorporated into a user-friendly computer program suite for extracting oligomer size and concentration from images of fluorescently tagged membrane proteins. This method, termed two-dimensional fluorescence intensity fluctuation (FIF) spectrometry, which can generate ‘spectrograms’ consisting of frequencies of occurrence of brightness-concentration pairs of values across pixels of fluorescence images of cells expressing molecules labeled with single fluorescent tags, and unmixes the spectrograms to determine the proportion of different oligomer sizes. The computer program suite is capable of producing a complete set of data per type of sample in less than one day using a standard personal computer.
Copyrighted Software available for no charge for first 15 days and at discounted rate for non-commercial use. Please contact us for commercial use. This technology – including experimental design, measurements, and data analysis – is also available as a service to commercial and non-commercial customers.
Dr. Valerica Raicu, Department of Physics, UW-Milwaukee
Please contact our office to share your business’ needs.