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Gradient shadow pattern reveals refractive index of liquid.

ABSTRACT: We propose a simple method that uses a gradient shadow pattern (GSP) to measure the refractive index nL of liquids. A light source generates a "dark-bright-dark" GSP when it is projected through through the back of a transparent, rectangular block with a cylindrical chamber that is filled with a liquid sample. We found that there is a linear relationship between nL and the proportion of the bright region in a GSP, which provides the basic principle of the proposed method. A wide range 1.33???nL???1.46 of liquids was measured in the single measurement setup with error <0.01. The proposed method is simple but robust to illuminating conditions, and does not require for any expensive or precise optical components, so we expect that it will be useful in many portable measurement systems that use nL to estimate attributes of liquid samples.

SUBMITTER: Kim W

PROVIDER: S-EPMC4908432 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Gradient shadow pattern reveals refractive index of liquid.

Kim Wonkyoung W Kim Dong Sung DS

Scientific reports 20160615

We propose a simple method that uses a gradient shadow pattern (GSP) to measure the refractive index nL of liquids. A light source generates a "dark-bright-dark" GSP when it is projected through through the back of a transparent, rectangular block with a cylindrical chamber that is filled with a liquid sample. We found that there is a linear relationship between nL and the proportion of the bright region in a GSP, which provides the basic principle of the proposed method. A wide range 1.33 ≤ nL ...[more]

PMID: 27302603

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Project description:Refractive index-based sensors offer attractive characteristics as nondestructive and universal detectors for liquid chromatographic separations, but a small dynamic range and sensitivity to minor thermal perturbations limit the utility of commercial RI detectors for many potential applications, especially those requiring the use of gradient elutions. As such, RI detectors find use almost exclusively in sample abundant, isocratic separations when interfaced with high-performance liquid chromatography. Silicon photonic microring resonators are refractive index-sensitive optical devices that feature good sensitivity and tremendous dynamic range. The large dynamic range of microring resonators allows the sensors to function across a wide spectrum of refractive indices, such as that encountered when moving from an aqueous to organic mobile phase during a gradient elution, a key analytical advantage not supported in commercial RI detectors. Microrings are easily configured into sensor arrays, and chip-integrated control microrings enable real-time corrections of thermal drift. Thermal controls allow for analyses at any temperature and, in the absence of rigorous temperature control, obviates extended detector equilibration wait times. Herein, proof of concept isocratic and gradient elution separations were performed using well-characterized model analytes (e.g., caffeine, ibuprofen) in both neat buffer and more complex sample matrices. These experiments demonstrate the ability of microring arrays to perform isocratic and gradient elutions under ambient conditions, avoiding two major limitations of commercial RI-based detectors and maintaining comparable bulk RI sensitivity. Further benefit may be realized in the future through selective surface functionalization to impart degrees of postcolumn (bio)molecular specificity at the detection phase of a separation. The chip-based and microscale nature of microring resonators also make it an attractive potential detection technology that could be integrated within lab-on-a-chip and microfluidic separation devices.

Project description:IntroductionOptical techniques are routinely used to size and count extracellular vesicles (EV). For comparison of data from different methods and laboratories, suitable calibrators are essential. A suitable calibrator must have a refractive index (RI) as close to that of EV as possible but the RI of EV is currently unknown. To measure EV, RI requires accurate knowledge of size and light scattering. These are difficult to measure as most EVs cannot be resolved by light microscopy and their diameter is smaller than the wavelength of visible light. However, nanoparticle tracking analysis (NTA) provides both size and relative light scattering intensity (rLSI) values. We therefore sought to determine whether it was possible to use NTA to measure the RI of individual EVs.MethodsNTA was used to measure the rLSI and size of polystyrene and silica microspheres of known size and RI (1.470 and 1.633, respectively) and of EV isolated from a wide range of cells. We developed software, based on Mie scattering code, to calculate particle RI from the rLSI data. This modelled theoretical scattering intensities for polystyrene and silica microspheres of known size (100 and 200 nm) and RI. The model was verified using data from the polystyrene and silica microspheres. Size and rLSI data for each vesicle were processed by the software to generate RI values.ResultsThe following modal RI measurements were obtained: fresh urinary EV 1.374, lyophilised urinary EV 1.367, neuroblastoma EV 1.393, blood EV 1.398, EV from activated platelets 1.390, small placental EV 1.364-1.375 and 1.398-1.414 for large placental EV (>200 nm). Large placental EV had a significantly higher RI than small placental EV (p<0.0001). The spread of RI values was narrower for small EV than for the more heterogeneous large EV.DiscussionUsing NTA and Mie scattering theory, we have demonstrated that it is possible to estimate the RI of sub-micron EV using NTA data. EV typically had a modal RI of 1.37-1.39, whereas values of >1.40 were observed for some large (>200 nm) microvesicles.ConclusionThis method for measuring EV RI will be useful for developing appropriate calibrators for EV measurement.

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Gradient shadow pattern reveals refractive index of liquid.

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Gradient shadow pattern reveals refractive index of liquid.

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