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A possible pathogenetic factor of sickle-cell disease based on fluorescent analysis via an optofluidic resonator.

ABSTRACT: Waveguide based optofluidic resonator features high precision and high sensitivity in real-time fluorescent analysis. We present a novel optofluidic resonator following the hollow-core metal-cladding waveguide structure, which is then used to record the real-time binding process of Fe²⁺ and Fe³⁺ with protoporphyrin IX (PpIX) in PBS solution, respectively. The central fluorescent wavelength of compound with Fe²⁺ is in good accordance with that of the normal hemoglobin, whilst the peaks of the Fe³⁺ compound match the hemoglobin specimen from sickle-cell disease (SCD) patients. Similar statement holds when we monitor the real-time oxidation processes of these products by injecting oxygen into the optofluidic chip. These observations lead to the speculation that the SCD is caused by replacing the Fe²⁺ in hemoglobin with Fe³⁺, which may be insightful in the discovery of new clinical routes to cure this disease.

SUBMITTER: Dai H

PROVIDER: S-EPMC5466609 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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A possible pathogenetic factor of sickle-cell disease based on fluorescent analysis via an optofluidic resonator.

Dai Hailang H Yin Cheng C Ye Xiaona X Jiang Bei B Ran Maowu M Cao Zhuangqi Z Chen Xianfeng X

Scientific reports 20170609 1

Waveguide based optofluidic resonator features high precision and high sensitivity in real-time fluorescent analysis. We present a novel optofluidic resonator following the hollow-core metal-cladding waveguide structure, which is then used to record the real-time binding process of Fe2+ and Fe3+ with protoporphyrin IX (PpIX) in PBS solution, respectively. The central fluorescent wavelength of compound with Fe2+ is in good accordance with that of the normal hemogl ...[more]

PMID: 28600527

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Project description:Fluorescent microscopy and flow cytometry are widely used tools in biomedical sciences. Cost-effective translation of these technologies to remote and resource-limited environments could create new opportunities especially for telemedicine applications. Toward this direction, here we demonstrate the integration of imaging cytometry and fluorescent microscopy on a cell phone using a compact, lightweight, and cost-effective optofluidic attachment. In this cell-phone-based optofluidic imaging cytometry platform, fluorescently labeled particles or cells of interest are continuously delivered to our imaging volume through a disposable microfluidic channel that is positioned above the existing camera unit of the cell phone. The same microfluidic device also acts as a multilayered optofluidic waveguide and efficiently guides our excitation light, which is butt-coupled from the side facets of our microfluidic channel using inexpensive light-emitting diodes. Since the excitation of the sample volume occurs through guided waves that propagate perpendicular to the detection path, our cell-phone camera can record fluorescent movies of the specimens as they are flowing through the microchannel. The digital frames of these fluorescent movies are then rapidly processed to quantify the count and the density of the labeled particles/cells within the target solution of interest. We tested the performance of our cell-phone-based imaging cytometer by measuring the density of white blood cells in human blood samples, which provided a decent match to a commercially available hematology analyzer. We further characterized the imaging quality of the same platform to demonstrate a spatial resolution of ~2 ?m. This cell-phone-enabled optofluidic imaging flow cytometer could especially be useful for rapid and sensitive imaging of bodily fluids for conducting various cell counts (e.g., toward monitoring of HIV+ patients) or rare cell analysis as well as for screening of water quality in remote and resource-poor settings.

Project description:Wolman disease in an inherited metabolic disease, characterized by a severe deficiency of the acid lipase and a massive lysosomal storage of triacylglycerols and cholesteryl esters, associated with hepatosplenomegaly, adrenal calcification and nearly always fatal in the first year of life. Cultured human lymphoblastoid cells and human adrenal cells are able to promote the formation of mildly oxidized low-density lipoproteins (LDL), which in turn exhibit a non-negligible cytotoxic effect on these cells. In contrast, fibroblasts induce only very low levels of LDL oxidation. Comparative experiments have shown that the cytotoxic effect of oxidized LDL was higher to Wolman-disease cells than to controls. The oxidative ability of Wolman cells was similar to that of normal ones. The over-cytotoxicity of mildly oxidized LDL to Wolman cells resulted from the higher uptake of mildly oxidized LDL through the LDL-receptor pathway, which is only poorly down-regulated in Wolman cells subsequently to the block of the lysosomal degradation of LDL-cholesteryl esters. In cultured adrenal cells, oxidized LDL induced a sustained rise in intracellular [Ca2+] which is directly involved in the cellular damage and cell death induced by oxidized LDL [Nègre-Salvayre and Salvayre (1992) Biochim. Biophys. Acta 1123, 207-215]. This Ca2+ peak is followed by a dramatic deposition of calcium in damaged or/and dead cultured adrenal cells, quite similar to that observed in Wolman-disease adrenal cortex. The cell-induced LDL oxidation and the subsequent cytotoxic effect can be prevented, at least in part, by antioxidants such as alpha-tocopherol and nordihydroguaiaretic acid. These findings support the hypothesis that the Wolman-disease adrenal damage (necrosis and calcification) could result from the association of the following events: mild oxidation of LDL by adrenal cells, over-uptake of mildly oxidized LDL by Wolman cells (resulting from the block of the lysosomal degradation of cholesteryl esters in Wolman cells), and cytotoxicity related to the amount of mildly oxidized LDL internalized by cells. The reported data also suggest that LDL oxidation induced by adrenal cells and their subsequent cytotoxicity can be prevented (in part) by antioxidants, and the potential therapeutic use of antioxidants in Wolman disease is discussed.

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A possible pathogenetic factor of sickle-cell disease based on fluorescent analysis via an optofluidic resonator.

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A possible pathogenetic factor of sickle-cell disease based on fluorescent analysis via an optofluidic resonator.

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