A spatial resolution evaluation method of endoscopic optical coherence tomography system using the annular phantom.

As an important biomedical imaging method, endoscopic optical coherence tomography (OCT) is necessary to check its performance regularly. The ordinary plane phantoms are only able to evaluate part of image tangent to the probe. In this research, a spatial resolution estimate method of the endoscope OCT system is proposed. The annular phantom, made by uniformly distributing golden scattered microparticles in polydimethylsiloxane (PDMS), can provide dynamic scanning imaging evaluation of endoscopic OCT system, closer to its actual working status. The point spread function analysis method is used to analyze the imaging results of the annular phantom with the endoscopic OCT system. And many scattered particles are statistically analyzed to determine the spatial resolution of the endoscope OCT system. The method is low in cost, simple and convenient. It is valuable for the development of test standards for endoscope OCT systems.

Measurement of the refractive index of whole blood and its components for a continuous spectral region.

The refractive index of blood is a key biophysical parameter, which can reflect the physiological state. We measured the refractive index of whole blood and other components, such as serum, plasma, and hemoglobin, based on internal reflection by using a homemade apparatus in the spectral range of 400 to 750 nm. In addition to the hemoglobin solution, which has a Soret band about 420 nm and two Q-bands between 500 and 600 nm, the measurements of other samples are the normal dispersion curve. The results are approximated by the Cauchy equation and Sellmeier equation, and the correlation coefficients are more than 0.997.

Complex Refractive Index Dispersion of Strong Absorbing Material Determined Using Internal Reflectance Spectra Measurement.

Complex refractive index dispersion (CRID) of offset inks is an important spectral property that affects the quality of printing. Due to the strong absorption of offset inks, great difficulty exists when measuring their CRID. In this study, a recently proposed apparatus that can detect the internal reflectance spectra was used to measure the CRID of three strong absorbing offset inks (magenta, yellow, and cyan). Both anomalous dispersion curve and extinction coefficient curve were well determined over the spectral range of 400-750 nm. This study experimentally proves that the apparatus and related method are feasible for the CRID measurement of strong absorbing materials and could serve as a powerful measuring tool for optical parameters.

Study of the inhibition effect of thiazone on muscle optical clearing.

We investigated the effect of thiazone, a widely used penetration enhancer, on in vitro porcine skin and muscle tissue by single-integrating sphere technique during optical clearing (OC) treatment. The results showed that thiazone induced an increase on the total transmittance of skin which led to a reduction in that of muscle in the spectral range from 400 to 800 nm. Small particles crystalized out from the thiazone-treated muscle were observed by microscopy imaging. With the help of x-ray diffraction measurement, we ascertained that the crystal was a single-crystal of thiazone, which mainly induced an increase of the scattering. Contrast transmittance measurements carried on the mixture of water, thizaone­propylene glycol solution showed that the free water in muscle could be the main reason for the thiazone crystallization. Therefore, during OC treatment of thiazone, the remarkable effect on skin and the noticeable inhibition effect on subcutaneous muscle tissue after penetrating into the skin should be considered. The experimental results provide such a reference for the choice of penetration enhancer.

Quantitative index imaging of coculture cells by scanning focused refractive index microscopy.

We report the quantitative refractive index (RI) imaging of cocultured cells in their living environment by scanning focused refractive index microscopy (SFRIM). Mouse microglial cells and synovial cells are cocultured on the top surface of a trapezoid prism. The RI imaging of living cells is obtained in a reflection-type method. The RI information is deduced with the simple derivative total internal reflection method, where a complex retrieval algorithm or reconstruction process is unnecessary. The outline of each cell is determined according to the RI value compared with that of the immersion liquid. The cocultured cells can be discriminated in the RI image. The measurement is nondestructive and label-free. The experimental results prove that SFRIM is a promising tool in the field of biological optics.

Measurement of complex refractive index of turbid media by scanning focused refractive index.

We present the application of scanning focused refractive index microscopy in the complex refractive index measurement of turbid media. An extra standard scattering layer is placed in front of the detector to perform scattering transformation on the reflected light. The principle of the scattering transformation is elaborated theoretically. The influence of the sample scattering is deeply and effectively suppressed experimentally. As a proof of the feasibility and accuracy of the proposed method, we demonstrate experimental data of 20% and 30% Intralipid solutions that are commonly used as phantom media for light propagation studies.

Analysis of reflectance curve of turbid media and determination of the non-surface complex refractive index.

Based on the theory of Goos-Hänchen shift and its continuity near the critical angle, we introduce the concept of penetration depth below the critical angle, and obtain the general formula of reflectance using the gradient complex refractive index multilayered model. Compared with the fitting curve with Fresnel's Formula, our calculated results are more consistent with experimental results of Intralipid solution and the suspension of rutile TiO(2) powder. Combining the change of penetration depth near the critical angle with our model, we also reveal the essence of a simple method used to obtain the non-surface complex refractive index of turbid media.

Measurement of the refractive index of hemoglobin solutions for a continuous spectral region.

Determination of the refractive index of hemoglobin solutions over a wide wavelength range remains challenging. A famous detour approach is the Kramers-Kronig (KK) analysis which can resolve the real part of complex refractive index from the imaginary part. However, KK analysis is limited by the contradiction between the requirement of semi-infinite frequency range and limited measured range. In this paper, based on the Multi-curve fitting method (MFM), continuous refractive index dispersion (CRID) of oxygenated and deoxygenated hemoglobin solutions are measured using a homemade symmetrical arm-linked apparatus in the continuous wavelength range with spectral resolution of about 0.259nm. A novel method to obtain the CRID is proposed.

Continuous refractive index dispersion measurement based on derivative total reflection method.

Traditionally, continuous refractive index dispersion (CRID) measurement of materials with scattering is hard to realize. In this paper, CRID measurement based on the derivative total reflection method (CRIDM-DTRM) is proposed to measure the CRID of both absorption and scattering materials. It effectively determined the CRID of K9 glass, concentrated milk, and 0.5% methyl red solution in the 400-750 nm range with the spectral resolution of about 0.259 nm. For the first time, CRID of a scattering material is measured. CRIDM-DTRM is a useful technique in the field of RID measurement, especially for biotissues and anomalous dispersion materials.

Research on the change of complex refractive index of porcine muscle during natural dehydration.

The physical changes of tissue are complicated to evaluate during optical clearing (OC) treatment. Monitoring the changes of optical parameters, including the complex refractive index (CRI), helps people better understand the OC process. From the imaginary part of CRI, we can deduce the extinction coefficient of tissue. Based on the total internal reflection method, the time-dependent CRI of porcine muscle during natural dehydration is well determined. Results show that the real RI increases continuously with the increase of dehydration time, whereas the extinction coefficient initially increases and then decreases. Finally, the extinction coefficient becomes much smaller than the initial value, which demonstrates that better tissue optical clarity is obtained. The change tendency of the extinction coefficient of tissue is used to qualitatively explain the dynamic change of transmittance of a natural dehydrated tissue. Consequently, CRI, especially its imaginary part, is a very useful optical parameter by which to evaluate the OC effect.

Effect of the gradient of complex refractive index at boundary of turbid media on total internal reflection.

We analyze the existence of non-uniformity at the boundary of turbid media, and develop a gradient complex refractive index multilayered model in terms of this fact. Our model reveals the physics mechanism of the discrepancies between experimental data above the critical angle and the fitting curve with Fresnel's Formula. Also, from the perspective of the energy flow, reflectance R is obtained by the simplified models. We get complex refractive indexes and reflectance curves by fitting experimental data of 20% and 30% Intralipid solutions and rutile TiO2 powder suspension with two different methods. Compared with Fresnel's Formula, our model can fit experimental data better.

Two-dimensional scanning focused refractive-index microscopy and applications to refractive-index profiling of optical fibers.

The refractive-index profile (RIP) of optical fibers is of fundamental significance in determining critical fiber properties. Here, we present the application of a two-dimensional (2-D) scanning focused refractive-index microscopy (SFRIM) to accurately obtain the 2-D RIP of a graded-index optical fiber. Some modifications are made to SFRIM for better 2-D measurement. Quantitative RIP of the fiber is obtained with derivative total reflection method. The refractive-index accuracy is 0.002. The measured result is in good agreement with theoretical expectation. This method is straightforward, simple, repeatable, and free from signal distortion. This technique is suitable for symmetric and asymmetric optical fibers. The results indicate that this technique can be applied to obtain the RIPs of a wide range of materials and has broad application prospect in many fields.

Study on the refractive index matching effect of ultrasound on optical clearing of bio-tissues based on the derivative total reflection method.

In recent years, the tissue optical clearing (OC) technique in the biomedicine field has drawn lots of attention. Various physical and chemical methods have been introduced to improve the efficacy of OC. In this study, the effect of the combination of glycerol and ultrasound treatment on OC of in vitro porcine muscle tissues has been investigated. The refractive index (RI) matching mechanism of OC was directly observed based on the derivative total reflection method. A theoretical model was used to simulate the proportion of tissue fluid in the illuminated area. Moreover, the total transmittance spectra have been obtained by a spectrometer over the range from 450 nm to 700 nm. The administration of glycerol and ultrasound has led to an increase of the RI of background medium and a more RI matching environment was achieved. The experimental results support the validity of the ultrasound treatment for OC. The RI matching mechanism has been firstly quantitatively analyzed based on the derivative total reflection method.

Scanning focused refractive-index microscopy.

We present a novel scanning focused refractive-index microscopy (SFRIM) technique to obtain the refractive index (RI) profiles of objects. The method uses a focused laser as the light source, and combines the derivative total reflection method (DTRM), projection magnification, and scanning technique together. SFRIM is able to determine RIs with an accuracy of 0.002, and the central spatial resolution achieved is 1 µm, which is smaller than the size of the focal spot. The results of measurements carried out on cedar oil and a gradient-refractive-index (GRIN) lens agree well with theoretical expectations, verifying the accuracy of SFRIM. Furthermore, using SFRIM, to the best of our knowledge we have extracted for the first time the RI profile of a periodically modulated photosensitive gelatin sample. SFRIM is the first RI profile-resolved reflected light microscopy technique that can be applied to scattering and absorbing samples. SFRIM enables the possibility of performing RI profile measurements in a variety of applications, including optical waveguides, photosensitive materials and devices, photorefractive effect studies, and RI imaging in biomedical fields.

Study of dynamic pressure-induced refractive index change using derivative total reflection method.

We report the dynamic refractive index (RI) change of tissue under a stepped compression load using a custom-built pressure apparatus. Angle-dependent reflectance profiles of biotissue samples are recorded, and the RI values are resolved using the derivative total reflection method. These results are relevant for understanding the mechanism of mechanical optical clearing, for investigating tissue dynamics under mechanical stimuli, and for other biomedical applications.