Yang-Hsiang Chan

Ratiometric Temperature Sensing with Semiconducting Polymer Dots

This communication describes ultrabright single-nanoparticle ratiometric temperature sensors based on semiconducting polymer dots (Pdots). We attached the temperature sensitive dye-Rhodamine B (RhB), whose emission intensity decreases with increasing temperature-within the matrix of Pdots. The as-prepared Pdot-RhB nanoparticle showed excellent temperature sensitivity and high brightness because it took advantage of the light harvesting and amplified energy transfer capability of Pdots. More importantly, the Pdot-RhB nanoparticle showed ratiometric temperature sensing under a single wavelength excitation and has a linear temperature sensing range that matches well with the physiologically relevant temperatures. We employed Pdot-RhB for measuring intracellular temperatures in a live-cell imaging mode. The exceptional brightness of Pdot-RhB allows this nanoscale temperature sensor to be used also as a fluorescent probe for cellular imaging.

Ultrasensitive Copper(II) Detection Using Plasmon-Enhanced and Photo-Brightened Luminescence of CdSe Quantum Dots

Here, we present a simple platform for the use of the enhanced emission of 16-mercaptohexadecanoic acid (16-MHA) capped CdSe quantum dots (QDs) as a probe for ultrasensitive copper(II) detection. In this study, the photoluminescence (PL) of the QDs was first enhanced by Ag nanoprisms which were self-assembled on Si surfaces and then further increased by photobrightening. Using this approach, the control and different analytes could be readily probed all on a single platform using fluorescence microscopy. The enhanced PL intensity of CdSe QDs was selectively quenched in the presence of Cu(2+), accompanied by the emergence of a new red-shifted luminescence band. The quenching mechanism was found to be due to a cation exchange mechanism as confirmed by X-ray photoelectron spectroscopy (XPS) measurements. Herein, we have demonstrated that this simple methodology can offer a rapid and reliable detection of Cu(2+) with a detection limit as low as 5 nM and a dynamic range up to 100 muM in a fixed fast reaction time of 5 min. The potential applications of this technique were tested in two ways, for mixed-ion solutions and in physiological fluids, and both experiments exhibited good selectivity toward Cu(2+).

Copper(II) and iron(II) ion sensing with semiconducting polymer dots

This communication describes a simple platform that employs carboxyl functionalized semiconducting polymer dots as a fluorescent probe for sensitive ratiometric Cu(2+) and Fe(2+) detection, in which the sensing mechanism is based on aggregation-induced fluorescence quenching.

Multicolor Fluorescent Semiconducting Polymer Dots with Narrow Emissions and High Brightness

Fluorescent semiconducting polymer dots (Pdots) have attracted great interest because of their superior characteristics as fluorescent probes, such as high fluorescence brightness, fast radiative rates, and excellent photostability. However, currently available Pdots generally exhibit broad emission spectra, which significantly limit their usefulness in many biological applications involving multiplex detections. Here, we describe the design and development of multicolor narrow emissive Pdots based on different boron dipyrromethene (BODIPY) units. BODIPY-containing semiconducting polymers emitting at multiple wavelengths were synthesized and used as precursors for preparing the Pdots, where intraparticle energy transfer led to highly bright, narrow emissions. The emission full width at half-maximum of the resulting Pdots varies from 40 to 55 nm, which is 1.5-2 times narrower than those of conventional semiconducting polymer dots. BODIPY 520 Pdots were about an order of magnitude brighter than commercial Qdot 525 under identical laser excitation conditions. Fluorescence imaging and flow cytometry experiments indicate that the narrow emissions from these bright Pdots are promising for multiplexed biological detections.

Hybrid Semiconducting Polymer Dot–Quantum Dot with Narrow-Band Emission, Near-Infrared Fluorescence, and High Brightness

This communication describes a new class of semiconducting polymer nanoparticle-quantum dot hybrid with high brightness, narrow emission, near-IR fluorescence, and excellent cellular targeting capability. Using this approach, we circumvented the current difficulty with obtaining narrow-band-emitting and near-IR-fluorescing semiconducting polymer nanoparticles while combining the advantages of both semiconducting polymer nanoparticles and quantum dots. We further demonstrated the use of this new class of hybrid nanomaterial for effective and specific cellular and subcellular labeling without any noticeable nonspecific binding. This hybrid nanomaterial is anticipated to find use in a variety of in vitro and in vivo biological applications.

Using Patterned Arrays of Metal Nanoparticles to Probe Plasmon Enhanced Luminescence of CdSe Quantum Dots

Here we present a simple platform for probing plasmon enhanced photoluminescence (PL) of quantum dots by confocal microscopy. In this study, self-assembled monolayers of silane-derivative molecules were patterned onto the oxidized GaAs surfaces to direct the attachment of Au or Ag nanoparticles onto the surface. Following the directed binding of metal nanoparticles (MNPs), a layer-by-layer deposition of oppositely charged polymers was used to create films with varying thickness by controlling the numbers of deposited layers. CdSe quantum dots (QDs) of ∼4 and 5.5 nm in diameter with 16-mercaptohexadecanoic acid as a surfactant were then adsorbed onto the outermost polymer layer via electrostatic interactions. Using confocal fluorescence microscopy, the enhanced PL from the CdSe over the Au or Ag nanoparticle patterns could be imaged directly and scaled against the regions with no Au or Ag nanoparticles, and the luminescence of the GaAs (as an internal standard) for different CdSe-metal separations. By using a pattern, PL enhancement as a function of particle-CdSe spacing can be readily probed all on a single platform, where the QDs over MNPs and not over MNPs can be directly compared in the same dielectric environment. The observed luminescence as a function of metal-QD separation can be readily fit to a combined model of metal-fluorophore fluorescence quenching and local electric field enhancement.

A Compact and Highly Fluorescent Orange-Emitting Polymer Dot for Specific Subcellular Imaging

We demonstrate a new compact CN-PPV dot, which emits in the orange wavelength range with high brightness. The small particle size, high brightness, and the ability to highly specifically target subcellular structures make the CN-PPV dots promising probes for biological imaging and bioanalytical applications.

Squaraine-Based Polymer Dots with Narrow, Bright Near-Infrared Fluorescence for Biological Applications

This article describes the design and development of squaraine-based semiconducting polymer dots (Pdots) that show large Stokes shifts and narrow-band emissions in the near-infrared (NIR) region. Fluorescent copolymers containing fluorene and squaraine units were synthesized and used as precursors for preparing the Pdots, where exciton diffusion and likely through-bond energy transfer led to highly bright and narrow-band NIR emissions. The resulting Pdots exhibit the emission full width at half-maximum of ∼36 nm, which is ∼2 times narrower than those of inorganic quantum dots in the same wavelength region (∼66 nm for Qdot705). The squaraine-based Pdots show a high fluorescence quantum yield (QY) of 0.30 and a large Stokes shift of ∼340 nm. Single-particle analysis indicates that the average per-particle brightness of the Pdots is ∼6 times higher than that of Qdot705. We demonstrate bioconjugation of the squaraine Pdots and employ the Pdot bioconjugates in flow cytometry and cellular imaging applications. Our results suggest that the narrow bandwidth, high QY, and large Stokes shift are promising for multiplexed biological detections.

Quinoxaline-Based Polymer Dots with Ultrabright Red to Near-Infrared Fluorescence for In Vivo Biological Imaging

This article describes the design and synthesis of quinoxaline-based semiconducting polymer dots (Pdots) that exhibit near-infrared fluorescence, ultrahigh brightness, large Stokes shifts, and excellent cellular targeting capability. We also introduced fluorine atoms and long alkyl chains into polymer backbones and systematically investigated their effect on the fluorescence quantum yields of Pdots. These new series of quinoxaline-based Pdots have a fluorescence quantum yield as high as 47% with a Stokes shift larger than 150 nm. Single-particle analysis reveals that the average per-particle brightness of the Pdots is at least 6 times higher than that of the commercially available quantum dots. We further demonstrated the use of this new class of quinoxaline-based Pdots for effective and specific cellular and subcellular labeling without any noticeable nonspecific binding. Moreover, the cytotoxicity of Pdots were evaluated on HeLa cells and zebrafish embryos to demonstrate their great biocompatibility. By taking advantage of their extreme brightness and minimal cytotoxicity, we performed, for the first time, in vivo microangiography imaging on living zebrafish embryos using Pdots. These quinoxaline-based NIR-fluorescent Pdots are anticipated to find broad use in a variety of in vitro and in vivo biological research.

Stable Functionalization of Small Semiconducting Polymer Dots via Covalent Cross‐Linking and Their Application for Specific Cellular Imaging

A facile cross-linking strategy covalently links functional molecules to semiconducting polymer dots (Pdots) while simultaneously providing functional groups for biomolecular conjugation. In addition to greatly enhanced stability, the formed Pdots are small (<10 nm), which can be difficult to achieve with current methods but is highly desirable for most biological applications. These characteristics are significant for improving labeling efficiency and sensitivity in cellular assays that employ Pdots.