RESUMO
The Nobel Prize in Chemistry was awarded in 2000 for the discovery of conductive organic polymers, which have subsequently been adapted for applications in ultrasensitive biological detection. Here, we report the first use of this new class of fluorescent probes in a diverse range of cytometric and imaging applications. We demonstrate that these "Brilliant Violet" reporters are dramatically brighter than other UV-violet excitable dyes, and are of similar utility to phycoerythrin (PE) and allophycocyanin (APC). They are thus ideally suited for cytometric assays requiring high sensitivity, such as MHC-multimer staining or detection of intracellular antigens. Furthermore, these reporters are sensitive and spectrally distinct options for fluorescence imaging, two-photon microscopy and imaging cytometry. These ultra-bright materials provide the first new high-sensitivity fluorescence probes in over 25 years and will have a dramatic impact on the design and implementation of multicolor panels for high-sensitivity immunofluorescence assays.
Assuntos
Imunofluorescência/métodos , Corantes Fluorescentes/análise , Polímeros/análise , Coloração e Rotulagem/métodos , Animais , Encéfalo/ultraestrutura , Condutividade Elétrica , Citometria de Fluxo , Corantes Fluorescentes/química , Humanos , Leucócitos Mononucleares/citologia , Camundongos , Microscopia de Fluorescência por Excitação Multifotônica , Imagem Molecular , Ficocianina/análise , Ficoeritrina/análise , Polímeros/química , Retina/ultraestrutura , Sensibilidade e EspecificidadeRESUMO
The growth of films of [H2B(pz)2]Fe(ii)(bpy) on Au(111) is characterized from the bilayer film to multilayer film regime. Scanning tunneling microscopy shows a transition from a well-ordered, uniform bilayer film to a poorly-ordered film at larger thicknesses. Previous local tunneling spectroscopy and conductance mapping in bilayer films permit the identification of coexisting molecular spin-states at all temperatures. New ultraviolet photoelectron spectroscopy is consistent with this picture and in agreement with the density of states calculated by density functional theory. In thicker films with a polycrystalline morphology, evidence for a more bulk-like change in spin composition as a function of temperature is obtained by observing the reduction in intensity of Fe 2p core level satellites in X-ray photoelectron spectra.