Liqing Sun

Quantitative analysis of collagen change between normal and cancerous thyroid tissues based on SHG method

Second-harmonic generation (SHG) is proved to be a high spatial resolution, large penetration depth and non-photobleaching method. In our study, SHG method was used to investigate the normal and cancerous thyroid tissue. For SHG imaging performance, system parameters were adjusted for high-contrast images acquisition. Each x-y image was recorded in pseudo-color, which matches the wavelength range in the visible spectrum. The acquisition time for a 512×512-pixels image was 1.57 sec; each acquired image was averaged four frames to improve the signal-to-noise ratio. Our results indicated that collagen presence as determined by counting the ratio of the SHG pixels over the whole pixels for normal and cancerous thyroid tissues were 0.48±0.05, 0.33±0.06 respectively. In addition, to quantitatively assess collagen-related changes, we employed GLCM texture analysis to the SHG images. Corresponding results showed that the correlation both fell off with distance in normal and cancerous group. Calculated value of Corr50 (the distance where the correlation crossed 50% of the initial correlation) indicated significant difference. This study demonstrates that SHG method can be used as a complementary tool in thyroid histopathology.

Nasopharyngeal carcinoma detection by tissue smears using surface-enhanced Raman spectroscopy

ABSTRACT Nasopharyngeal carcinoma is one of the most common malignant neoplasms in head and neck. In this work, surface-enhanced Raman spectroscopy technique was used to study the molecular differences between cancerous and noncancerous smear samples which were obtained after clinical biopsy by smearing the tissue on the slides. Principal component analysis combined with linear discriminant analysis provided a sensitivity of 79.4% and a specificity of 81.8% for differentiation between cancerous and noncancerous nasopharyngeal tissue smears. This work provides a good basis for the methodology of nasopharyngeal tissue smear based on surface-enhanced Raman spectroscopy technique and is worth further studying.

Discrimination of Nasopharyngeal Carcinoma from Noncancerous Ex Vivo Tissue Using Reflectance Spectroscopy

Reflectance spectroscopy is a low-cost, nondestructive, and noninvasive method for detection of neoplastic lesions of mucosal tissue. This study aims to evaluate the capability of reflectance spectroscopy system under white light (400–700 nm) with a multivariate statistical analysis for distinguishing nasopharyngeal carcinoma (NPC) from nasopharyngeal benign ex vivo tissues. High quality reflectance spectra were acquired from nasopharyngeal ex vivo tissues belonging to 18 noncancerous and 19 cancerous subjects, and the combination of principal component analysis-linear discriminant analysis (PCA-LDA) along with leave-one-spectrum-out cross-validation (LOOCV) diagnostic algorithm was subsequently employed to classify different types of tissue group, achieving a diagnostic sensitivity of 73.7% and a specificity of 72.2%. Furthermore, in order to distinguish NPC from nasopharyngeal benign ex vivo tissues based on reflectance spectra simply, spectral intensity ratios of oxyhemoglobin (540/576) were used as an indicator of the carcinogenesis associated transformation in the hemoglobin oxygenation. This tentative work demonstrated the potential of reflectance spectroscopy for NPC detection using ex vivo tissue and has significant experimental and clinical value for further in vivo NPC detection in the future.

Surface-enhanced Raman spectroscopy of creatinine in silver colloid

Surface enhanced Raman spectroscopy (SERS) technology has already made great progress in bio-molecule detection. It can make the target molecules strongly absorbed onto the surface of metal nanoparticles, and then the Raman signal of its own has been greatly enhanced through physical and chemical enhancement mechanisms. We report the SERS spectra of creatinine in silver colloid, and study the silver colloid enhanced effects on the Raman scattering of creatinine. We can also find that creatinine concentration is linearly related to its SERS peak intensity and the detection limit of creatinine silver sol is found to be 10 mg/dl. In conclusion, we can observe that the silver colloid has very good enhanced effects for the creatinine. The potential applications of SERS in quantitative measurement of the creatinine liquor are demonstrated. The result shows that the SERS approach would provide a unique and fast test method for creatinine detection.

Diagnostic potential for gold nanoparticle-based surface-enhanced Raman spectroscopy to provide colorectal cancer screening using blood serum sample

Surface-enhanced Raman spectroscopy (SERS) is a vibrational spectroscopic technique that is capable of probing the biomolecular changes associated with diseased transformation. The objective of our study was to explore gold nanoparticle based SERS to obtain blood serum biochemical information for non-invasive colorectal cancer detection. SERS measurements were performed on two groups of blood serum samples: one group from patients (n = 38) with pathologically confirmed colorectal cancer and the other group from healthy volunteers (control subjects, n = 45). Tentative assignments of the Raman bands in the measured SERS spectra suggested interesting cancer specific biomolecular changes, including an increase in the relative amounts of nucleic acid, a decrease in the percentage of saccharide and proteins contents in the blood serum of colorectal cancer patients as compared to that of healthy subjects. Principal component analysis (PCA) of the measured SERS spectra separated the spectral features of the two groups into two distinct clusters with little overlaps. Linear discriminate analysis (LDA) based on the PCA generated features differentiated the nasopharyngeal cancer SERS spectra from normal SERS spectra with high sensitivity (97.4%) and specificity (100%). The results from this exploratory study demonstrated that gold nanoparticle based SERS serum analysis combined with PCA-LDA has tremendous potential for the non-invasive detection of colorectal cancers.

Comparative analysis of diagnostic efficiency of confocal micro-Raman spectroscopy and surface-enhanced Raman spectroscopy for nasopharyngeal carcinoma detection based on tissue smears method

Raman spectroscopy is a practice tool for tissue detection. This work demonstrated that confocal micro-Raman spectroscopy (CMRS) and surface-enhanced Raman spectroscopy (SERS) have promising exploratory potential for the detection of nasopharyngeal carcinoma (NPC) based on nasopharyngeal swab smear.

Using micro-Raman spectroscopy for nasopharyngeal cancerous tissue detection

Micro-Raman spectroscopy is widely used for non-invasive tissue diagnosis and detection, as it provides detailed information about biomolecular composition, structure, and interaction of tissue. In this work, micro-Raman spectroscopy was used to investigate non-cancerous and cancerous nasopharyngeal tissues. The obtained nasopharyngeal tissue samples in vitro are divided into two groups: cancerous (n=12, undifferentiated non-keratinizing carcinomas) and non-cancerous (n=10, 7 chronic inflammations, 2 lymphomas and 1 lymphocytosis). Firstly, we analyzed the Raman spectra in the fingerprint (FP, 400-1800cm-1) region acquired. Preliminary results showed that there are some spectral differences in different pathological conditions. Furthermore, Raman spectra from cancerous and non-cancerous nasopharyngeal tissue in the high wavenumber region (HW, 2800-3100cm-1) were also reported for the first time. After detailed analysis, we achieved significant differences in Raman bands at 2854, 2874, 2934, and 3067cm-1 between cancerous and non-cancerous nasopharyngeal tissues. This study demonstrates that both fingerprint and high wavenumber regions of micro-Raman spectroscopy have the potential for the early detection of nasopharyngeal carcinomas.

Immunoassay for CEA using the novel probe-labeled Ag nanoparticles based on surface-enhanced Raman scattering

In this study, we report the Ag nanoparticles aggregated in the process of the labeling that use the crystal violet as the Raman probe. After this process- immune nanoparticles aggregate with high SERS sensitivity and biological specificity are created. We track and characterize the preparation by employing UV-Vi absorption spectra, Transmission Electron Microscope (TEM) and SERS spectra. With the increase of crystal violet, the aggregate of the Ag nanoparticles also increase, while the intensity of absorption peaks decrease. When the concentration of crystal violet reaches 1.0×10-4 mol/L, new peaks were found in the long wavelengths, and with the increase of the crystal violet, the intensity of the new peaks increase as well. We observe from the TEM, that with the increase of crystal violet, the aggregation degree of the Ag nanoparticles also increase and then they unite together. The SERS activity of the aggregates was directly related to the aggregation degree. Detecting the SERS activity of Ag NPs aggregates labeled with different amount of crystal violet, we found that with the increase of the crystal violet, the SERS signals of NPs aggregates enhanced. However, when the amount of crystal violet exceeded 25μL in 1mL colloidal silver, the Ag NPs occurred agglomeration and thereafter the next preparation of immune-label aggregates was hindered. Whereas, the probe labelled with 12μL crystal violet exhibited a better stability, stronger SERS activity and higher biological specificity, and it may accomplish a highly efficient SERS-based immunoassay. This immune probe was applied for detecting the expression of carcinoembryonic antigen (CEA) in the colon cancer tissue slice. Results show that appropriate immune aggregates labelled with optimum quantity of crystal violet present high stability, strong SERS activity and good immune specificity, which are expected to be applied in the analysis of the protein expression in the tissue section and promising for developing into a clinical tool for diagnosis.

Study of ABO blood types by combining membrane electrophoresis with surface-enhanced Raman spectroscopy

The molecular characterization of ABO blood types, which is clinically significant in blood transfusion, has clinical and anthropological importance. Polymerase chain reaction sequence-based typing (PCR-SBT) is one of the most commonly used methods for the analysis of genetic bases of ABO blood types. However, such methods as PCR-SBT are time-consuming and are high in demand of equipments and manipulative skill. Here we showed that membrane electrophoresis based SERS method employed for studying the molecular bases of ABO blood types can provide rapidand easy-operation with high sensitivity and specificity. The plasma proteins were firstly purified by membrane electrophoresis and then mixed with silver nanoparticles to perform SERS detection. We use this method to classify different blood types, including blood type A (n=13), blood type B (n=9) and blood type O (n=10). Combination of principal component analysis (PCA) and liner discriminant analysis (LDA) was then performed on the SERS spectra of purified albumin, showing good classification results among different blood types. Our experimental outcomes represent a critical step towards the rapid, convenient and accurate identification of ABO blood types.

Microwave synthesis of Au nanoparticles as promising SERS substrates

A novel method for rapidly synthesized Au colloidal under microwave irradiation was present in this paper. Size of the Au nanoparticles varied from 10 nm to 60 nm along with varying mol fractions by chloroauric acid solution reduced with sodium citrate. The prepared Au nanoparticles were characterized by transmission electron microscope (TEM) and ultraviolet-visible (UV-Vis) spectrophotometer. It is found that the nanoparticle size and shape are highly dependent on the reaction time and the molar ratios of the reducing agent. By the SERS measurements of R6G, 4-MBA and Crystal violet, this Au colloid is shown to be an excellent SERS substrate with good stability. As the fabrication process of this SERS substrate is simple and inexpensive, this method may be used in large-scale preparation of substrates that can serve as an ideal SERS substrate in biomedical application.