Researchers have found that decreased levels of circulating citrulline could be an indicator of intestinal failure. Typically, this amino acid, which is produced by the intestinal mucosa cells, circulates in the blood at a physiological level of ∼40 μM. The current methodology for measuring this level involves the use of bulky equipment, such as mass spectroscopy and analysis at a central laboratory, which can delay diagnosis. Therefore, the current detection method is unsuited for routine monitoring at a doctor’s office. Our research group proposes the development of a point-of-care (POC) device to overcome this issue. The proposed device utilizes surface-enhanced Raman spectroscopy (SERS) coupled with a specifically designed aptamer, capable of binding to citrulline, conjugated to colloidal gold nanoparticles. The assay is then embedded within a vertical flow paper-fluidic platform as a deliverable at the POC, and a handheld Raman spectrometer (638-nm excitation) was used to interrogate the sample. Results showed good dynamic range and specificity with an average 73% decrease in SERS signal intensity with increasing concentrations of citrulline (0 to 50 μM) in phosphate-buffered saline compared to its controls: glycine, glutamine, histidine, and valine, which showed less than 10% average decrease in the presence of 200 μM of each analyte. Further, the limit of detection (LOD) within a chip was determined to be 0.56 μM, whereas the LOD across chips was below 10 μM.
Emerging technologies are enabling the feasibility of new types of point-of-care diagnostic devices. A portable, multimodal microscopy platform intended for use in remote diagnostic applications is presented. Use of such a system could bring high-quality microscopy to field use for diseases such as malaria, allowing better diagnostic and surveillance information to be gathered. The microscope was designed using off-the-shelf components and a manual filter selection to generate bright-field, fluorescent, and cross-polarized images of samples mounted to microscopy slides. Design parameters for the system are discussed, and characterization is performed using standardized imaging targets, multimodal phantoms, and blood smears simulating those used in malaria diagnosis. The microscope is shown to be able to image below element 9-3 of a 1951 U.S. Air Force target, indicating that the system is capable of resolving features <775 nm. Morphological indicators of
As a first step toward development of this system four colloidal nanoparticles, spherical gold, silver cubes, silica-gold nanoshells, and silver-gold nanocages have been analyzed for use in the POC system. Transmission electron microscopy (TEM) images have been taken of each nanoparticle to visualize the morphology of the nanoparticles, which is vital for SERS. Ultraviolet-visible (UV/Vis) spectroscopy was also collected to verify the extinction spectra for each nanoparticle was in resonance with the excitation wavelength. The nanoparticles were functionalized with mercaptobenzoic acid (MBA), a Raman reporter molecule, and SERS spectra were collected to determine which has better utility in a novel micro-to-nanochannel. The data showed that the silver nanocubes have a larger enhancement factor than the gold nanospheres, nanoshells, or nanocages. Currently, these nanocubes are being functionalized with the citulline for assessing the concentration sensitivity and dynamic range for ultimate use as a marker for radiation.
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