title: Confocal fluorescence polarization microscopy for linear unmixing of spectrally similar labels.
abstract: Studies of biological samples often call for simultaneous identification of multiple molecular or structural components. Multiple labelling fluorescence techniques are a powerful way of achieving this. However, the ability to distinguish a number of fluorescent probes unambiguously can be restricted by the fact that fluorescence spectra are generally broad and overlapping. Recently a technique known as linear unmixing has been combined with spectral imaging to discriminate between multiple fluorophores. In this study a scheme is proposed whereby fluorescence polarization information is used to expand the capability of the linear unmixing technique to accommodate additional fluorescent probes. As a proof-of-concept, it is shown that this polarization-based technique can be used to divide the signals generated by two spectrally similar fluorescent probes into their separate components.
authors.affiliation: Center for Atom Optics and Ultrafast Spectroscopy, Swinburne University of Technology, P.O. Box 218, Hawthorn, Victoria 3122, Australia.
authors.forename: Kiang Wei
authors.initials: KW
authors.lastname: Kho
authors.forename: Paul R
authors.initials: PR
authors.lastname: Stoddart
authors.forename: Martin
authors.initials: M
authors.lastname: Harris
authors.forename: Alex P
authors.initials: AP
authors.lastname: Mazzolini