High brightness, chemical and photostability, tunable characteristics, and spectral and surface properties are important attributes for nanoparticle probes designed for live cell imaging. We describe a class of nanoparticles for high-resolution imaging of O2 that consists of a substituted conjugated polymer (polyfluorene or poly(fluorene-alt-benzothiadiazole)) acting as a FRET antenna and a fluorescent reference with covalently bound phosphorescent metalloporphyrin (PtTFPP, PtTPTBPF). The nanoparticles prepared from such copolymers by precipitation method display stability, enhanced (>5-10 times) brightness under one- and two-photon excitation, compatibility with ratiometric and lifetime-based imaging modes, and low toxicity for cells. Their cell-staining properties can be modulated with positively and negatively charged groups grafted to the backbone. The "zwitter-ionic" nanoparticles show high cell-staining efficiency, while their cell entry mechanisms differ for the different 3D models. When injected in the bloodstream, the cationic and anionic nanoparticles show similar distribution in vivo. These features and tunable properties make the conjugated polymer based phosphorescent nanoparticles a versatile tool for quantitative O2 imaging with a broad range of cell and 3D tissue models.
A method to monitor the level of oxygen in microdroplets is presented. Optical sensor nanoparticles are dispersed in the aqueous phase of the microfluidic droplets for culturing bacteria. The oxygen sensor nanoparticles consist of phosphorescent indicator dye embedded in poly(styrene-block-vinylpyrrolidone) nanobeads. The nanoparticles are excitable by red light and emit in the near-infrared spectra region which minimizes background fluorescence from biological matter. The biocompatibility of the nanoparticles was proven. Nanoparticles sensors were read out by adapted miniaturized oxygen meters. The instruments can be easily integrated into the microfluidic system by placing it next to the tubing and measuring through the tubing wall. The phosphorescence lifetime-based measurement circumvents the drawbacks of intensity-based measurements and enables the determination of the absolute oxygen concentration in individual moving droplets. The technique can also be used for monitoring the growth of bacteria in microdroplets. We demonstrate simultaneous measurement of concentration of oxygen and optical density (OD) from micro cultures of E. coli and M. smegmatis.
This review gives an overview on the state-of-the-art of oxygen imaging in microfluidics. Oxygen imaging using optical oxygen sensors based on luminescence is a versatile and powerful tool for obtaining profoundly space-resolved information of oxygen in microreactors and microfluidic systems. We briefly introduce the principle of oxygen imaging and present techniques of oxygen imaging applied in microreactors and microfluidic devices, including selection criteria and demands of sensing material and basic set-up for a 2D oxygen sensing system. A detailed review of oxygen imaging in microreactors and microfluidic systems is given on different applications in oxygen gradient monitoring, cell culturing, single-cell analysis and chemical reactions. Finally, we discuss challenges and trends of oxygen imaging in microfluidic systems.
The crystal structure of cadmium mercury thiocyanate [CdHg(SCN)4] is determined by means of a four circle diffractometer. The structural features and properties of the crystal are described. Blue-violet light output by frequency doubling of a 809 nm GaAlAs laser diode using a cadmium mercury thiocyanate crystal device that is 3 mm thick at the phase matching angle: θ=47.7°, φ=0°, is realized at room temperature with input power lower than 200 mW. A 404.5 nm light output power of 1.8 mW is also measured for a 576 mW input power of the GaAlAs laser diode.
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