TY - JOUR A1 - Hoffmann, Katrin A1 - Behnke, Thomas A1 - Drescher, Daniela A1 - Kneipp, Janina A1 - Resch-Genger, Ute T1 - Near-infrared-emitting nanoparticles for lifetime-based multiplexed analysis and imaging of living cells N2 - The increase in information content from bioassays and bioimaging requires robust and efficient strategies for the detection of multiple analytes or targets in a single measurement, thereby addressing current health and security concerns. For fluorescence techniques, an attractive alternative to commonly performed spectral or color multiplexing presents lifetime multiplexing and the discrimination between different fluorophores based on their fluorescence decay kinetics. This strategy relies on fluorescent labels with sufficiently different lifetimes that are excitable at the same wavelength and detectable within the same spectral window. Here, we report on lifetime multiplexing and discrimination with a set of nanometer-sized particles loaded with near-infrared emissive organic fluorophores chosen to display very similar absorption and emission spectra, yet different fluorescence decay kinetics in suspension. Furthermore, as a first proof-of-concept, we describe bioimaging studies with 3T3 fibroblasts and J774 macrophages, incubated with mixtures of these reporters employing fluorescence lifetime imaging microscopy. These proof-of-concept measurements underline the potential of fluorescent nanoparticle reporters in fluorescence lifetime multiplexing, barcoding, and imaging for cellular studies, cell-based assays, and molecular imaging. KW - Fluorescence lifetime imaging microscopy KW - FLIM KW - Lifetime multiplexing KW - Near infrared KW - NIR KW - Cell imaging KW - Nanoparticles PY - 2013 U6 - https://doi.org/10.1021/nn4029458 SN - 1936-0851 VL - 7 IS - 8 SP - 6674 EP - 6684 PB - ACS Publ. CY - Washington, DC, USA AN - OPUS4-29031 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hoffmann, Katrin A1 - Behnke, Thomas A1 - Drescher, Daniela A1 - Kneipp, Janina A1 - Resch-Genger, Ute T1 - Lifetime-based discrimination between spectrally matching vis and NIR emitting particle labels and probes N2 - Increasing the information content from bioassays which requires robust and efficient strategies for the detection of multiple analytes or targets in a single measurement is an important field of research, especially in the context of meeting current security and health concerns. An attractive alternative to spectral multiplexing, which relies on fluorescent labels excitable at the same wavelength, yet sufficiently differing in their emission spectra or color presents lifetime multiplexing. For this purpose, we recently introduced a new strategy based on 'pattern-matching' in the lifetime domain, which was exemplary exploited for the discrimination between organic dyes and quantum dot labels revealing multi-exponential decay kinetics and allowed quantification of these labels. Meanwhile, we have succeeded in extending this lifetime multiplexing approach to nanometer-sized particle labels and probes absorbing and emitting in the visible (vis) and near-infrared (NIR) spectral region. Here, we present a first proof-of-principle of this approach for a pair of NIR-fluorescent particles. Each particle is loaded with a single organic dye chosen to display very similar absorption and emission spectra, yet different fluorescence decay kinetics. Examples for the lifetime-based distinction between pairs of these fluorescent nanoparticles in solution and in cells are presented. The results underline the potential of fluorescenc lifetime multiplexing in life science and bioanalysis. KW - Fluorescence KW - Fluorescence lifetime imaging microscopy KW - FLIM KW - Lifetime Multiplexing KW - Particle Label KW - Near-infrared KW - NIR KW - Cell imaging KW - Nanoparticles PY - 2011 U6 - https://doi.org/10.1117/12.881442 SN - 1605-7422 VL - 7905 SP - 79051F-1 EP - 79051F-9 PB - SPIE, The International Society for Optical Engineering CY - Bellingham, Wash. AN - OPUS4-23637 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -