A simple inexpensive light-emitting diode (LED)-based fluorescence detector for detection in capillary separations is described. The modular design includes a separate high power LED source, detector head, designed in the epifluorescence arrangement, and capillary detection cells. The detector head and detection cells were printed using a 3D printer and assembled with commercially available optical components. Optical fibers were used for connecting the detector head to the LED excitation source and the photodetector module. Microscope objective or high numerical aperture optical fiber were used for collection of the fluorescence emission from the fused silica separation capillary. As an example, mixture of oligosaccharides labeled by 8-aminopyrene-1,3,6-trisulfonate (APTS) was separated by capillary zone electrophoresis and detected by the described detector. The performance of the detector was compared with both a semiconductor photodiode and photomultiplier as light sensing elements. The main advantages of the 3D printed parts, compared to the more expensive alternatives from the optic component suppliers, include not only cost reduction, but also easy customization of the spatial arrangement, modularity, miniaturization, and sharing of information between laboratories for easy replication or further modifications of the detector. All information and files necessary for printing the presented detector are enclosed in the Supporting Information.
The use of high quality fused silica capillary nanospray tips is critical for obtaining reliable and reproducible electrospray/MS data; however, reproducible laboratory preparation of such tips is a challenging task. In this work, we report on the design and construction of low-cost grinding device assembled from 3D printed and commercially easily available components. Detailed description and characterization of the grinding device is complemented by freely accessible files in stl and skp format allowing easy laboratory replication of the device. The process of sharpening is aimed at achieving maximal symmetricity, surface smoothness and repeatability of the conus shape. Moreover, the presented grinding device brings possibility to fabricate the nanospray tips of desired dimensions regardless of the commercial availability. On several samples of biological nature (reserpine, rabbit plasma, and the mixture of three aminoacids), performance of fabricated tips is shown on CE coupled to MS analysis. The special interest is paid to the effect of tip sharpness.
Water-soluble CdTe quantum dots (QDs) and their conjugates with antibodies and antigenes were prepared by optimized procedures for applications in CE immunoassays. The QD size of 3.5 nm, excitation spectrum in the range of 300-500 nm, the maximum wavelength of the emission spectrum at 610 nm, quantum yield of 0.25 and luminescence lifetimes in the range of 3.6-43 ns were determined. The 0.1 M solution of TRIS/TAPS (pH 8.3) was found to be the optimum buffer for the separation of the antiovalbumin-ovalbumin immunocomplex from the free conjugates of QDs.
Surface-enhanced Raman spectroscopy (SERS) is an extremely powerful analytical tool, which not only yields information about the molecular structure of the analyte in the form of characteristic vibrational spectrum but also gives sensitivities approaching those in fluorescence spectroscopy. The SERS measurement on the microfluidic platform provides possibility to manufacture the device with design perfectly fulfilling the needs of the application with minimal sample consumption. This review aims at describing basic strategies for SERS measurement in microfluidic devices published in the last decade and covers current trends in microfluidics with SERS detection in the field of bioanalysis and approaches toward on-line coupling of liquid-based separation techniques with SERS detection.
Resistive pulse sensing is a well‐known and established method for counting and sizing particles in ionic solutions. Throughout its development the technique has been expanded from detection of biological cells to counting nanoparticles and viruses, and even registering individual molecules, e.g., nucleotides in nucleic acids. This technique combined with microfluidic or nanofluidic systems shows great potential for various bioanalytical applications, which were hardly possible before microfabrication gained the present broad adoption. In this review, we provide a comprehensive overview of microfluidic designs along with electrode arrangements with emphasis on applications focusing on bioanalysis and analysis of single cells that were reported within the past five years.
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