Photoacoustic spectroscopy is a well-known spectroscopic method for gas detection. Exhaled breath contains volatile organic compounds (VOCs) as bio-markers of specific health condition. Detection and quantification of VOC bio-markers from exhaled breath can provide valuable information about the health status. We report the design and development of Photoacoustic spectroscopy setup and a preliminary study of the detection of certain standard VOCs.
Research in the field of laser plasma spectroscopy is gaining more importance due to its wide applications in various fields like astronomy, industry, environmental related studies, archeology and many more. The spectroscopic characterization of plasma generated in the meteorite samples during the Laser-induced Breakdown Spectroscopy (LIBS) shows the emission spectrum of elements present in it and also allows one to rapidly identify the elemental composition without any sample preparation and with good accuracy compared to some of the other methods adapted for the said purpose. Also, LIBS has other advantages like multi-elemental response, micro-nano gram level of destructiveness and portability of the instrument. The elemental analysis results for a few of the selected samples such as Iron meteorites, Lunar meteorites, Eucrites and impact glass are presented and discussed in this work. The results reveal that LIBS can be a potential alternative method for the meteorite fragment analysis.
Hyperspectral imaging (HSI), also called imaging spectrometer, originated from remote sensing. Hyperspectral imaging is an emerging imaging modality for medical applications, especially in disease diagnosis and image-guided surgery. HSI acquires a three-dimensional dataset called hypercube, with two spatial dimensions and one spectral dimension. Spatially resolved spectral imaging obtained by HSI provides diagnostic information about the objects physiology, morphology, and composition. The present work involves testing and evaluating the performance of the hyperspectral imaging system. The methodology involved manually taking reflectance of the object in many images or scan of the object. The object used for the evaluation of the system was cabbage and tomato. The data is further converted to the required format and the analysis is done using machine learning algorithm. The machine learning algorithms applied were able to distinguish between the object present in the hypercube obtain by the scan. It was concluded from the results that system was working as expected. This was observed by the different spectra obtained by using the machine-learning algorithm.
Urinary tract infection (UTI) is one of the major clinical problems known to mankind, especially among adult women.
Conventional methods for identification of UTI causing bacteria are time consuming and expensive. Therefore, a rapid
and cost-effective method is desired. In the present study, five bacteria (one Gram-positive and four Gram-negative),
most commonly known to cause UTI, have been identified and classified using micro-Raman spectroscopy combined
with principal component analysis (PCA).
Autofluorescence spectroscopy offer noninvasive and promising tools for the detection
of alternations biochemical compositions of tissues and cells, in presence of disease. They have
the added advantage of being highly objective due to the fact that diagnostic evaluation is by
statistical methods, eliminating errors from lack of experience, fatigue factor, and subjectivity of
visual perceptions. The present research work involves in designing and assembling of a low cost,
miniature oral cancer screening device with for routine clinical applications. A miniature system
was designed and assembled with much smaller and cost-effective components like compact light
source and miniature spectrometer, in a hand-held unit configuration. The performance of the
system was evaluated using animal -mouse- SCC model. The current system can be used in handheld
operation, which makes it very useful for many applications like, screening of squamous cell
carcinoma susceptible population.
Interaction of noble metal nanoparticles (NPs) with fluorophores has been an important research area in the field of material science and biomedical field. In the proximity of a metal nanoparticle, there is a quenching or enhancement in the intrinsic fluorescence of the fluorophore . The conditional quenching of the fluorescence can be used for negative sensing whereas enhancement in the fluorescence can be used to gain greater sensitivity and high signal to noise ratio in the molecular sensing/imaging. The current work deals with the systematic studies to understand the fluorescence quenching for few bio-fluorophores (NADH and FAD) when interacted with different sized silver nano-particles of (10nm, 40nm and 100nm). Home assembled Laser Induced Fluorescence (LIF) set-up was used to study the fluorescence quenching of NADH and FAD for different sized silver nanoparticles.
With a penchant for integrated photonics and miniaturization, the fabrication of micron sized optical elements using precision laser pulse management is drawing attention due to the possibility of minimizing tolerances for collateral material damage. The work presented here deals with the design, fabrication and characterization of a range of diffractive optics - gratings, grids and Fresnel zone plates – on transparent and metallic samples. Their low volume, light weight, transmission bandwidth, high damage threshold and flexible design make them suited for replacing conventional refractive optical elements. Our one-step, mask-less, 3-D laser direct writing process is a green fabrication technique which is in stark contrast to currently popular Photo-lithography based micro-structuring. Our method provides scope for modifications on the surface as well as within the bulk of the material. The mechanism involved in the fabrication of these optics on transparent and thin metallic substrates differ from each other. Our studies show that both amplitude and phase versions of micro-structures were achieved successfully with performances bearing ~98% accuracy vis-a-vis theoretical expectations.
Interest in the use of laser-based sensors operating in stand-off mode is increasing due to the wide range of options offered in the evaluation of distant targets. In this work, a stand-off laser induced breakdown spectroscopy (ST-LIBS) system has been developed and demonstrated to determine the heavy elements in soil samples. Initially, different parameters of the device have been optimized in ambient atmosphere and tested for quantitative analysis of Nickel and Chromium at two different stand-off distances (1 m and 6 m). The prepared concentrations were in the range of 100 ppm to 600 ppm. The laser energy of 150 mJ was maintained through out the experiment and to obtain a real time situation the target was in static state.
The ripening of fruits is associated with changes, in some cases subtle, in the color of the fruit. Traditionally spectroscopy used to measure these subtle changes and infer the ripeness of fruits. Spectrometers provides high-resolution but only measure a small area of the fruit. That might not be a good indicator of the overall ripeness. In this paper, we propose a compact tunable LED based hyper spectral imaging system that scans through a set of wavelengths and images, the reflectance from the whole fruit. Based on the type of fruit, only specific wavelengths need to be scanned. Following a validation using a Rubik’s cube, an example banana going through its ripening cycles is used to demonstrate the system.
Raman tweezers spectroscopy study of effect of free radicals was carried out on erythrocytes. We prepared hydroxyl radicals using Fenton reaction (which yields hydroxyl radicals). Raman spectra were acquired from single, trapped erythrocytes after supplementing with these free radicals. The changes in the Raman bands such as 1211 cm-1, 1224 cm-1, 1375 cm-1 indicate deoxygenation of red blood cells (RBCs). Our study shows that free radicals can induce oxidative stress on erythrocytes. The changes in the Raman spectra as well as shape of erythrocytes indicate that oxidative stress can trigger eryptosis in erythrocytes.
Laser Induced Breakdown Spectroscopy (LIBS) is a multi-elemental analysis technique with various advantages and has the ability to detect any element in real time. This technique holds a potential for environmental monitoring and various such analysis has been done in soil, glass, paint, water, plastic etc confirms the robustness of this technique for such applications. Compared to the currently available water quality monitoring methods and techniques, LIBS has several advantages, viz. no need for sample preparation, fast and easy operation, and chemical free during the process. In LIBS, powerful pulsed laser generates plasma which is then analyzed to get quantitative and qualitative details of the elements present in the sample. Another main advantage of LIBS technique is that it can perform in standoff mode for real time analysis. Water samples from industries and agricultural strata tend to have a lot of pollutants making it harmful for consumption. The emphasis of this project is to determine such harmful pollutants present in trace amounts in industrial and agricultural wastewater. When high intensity laser is made incident on the sample, a plasma is generated which gives a multielemental emission spectra. LIBS analysis has shown outstanding success for solids samples. For liquid samples, the analysis is challenging as the liquid sample has the chances of splashing due to the high energy of laser and thus making it difficult to generate plasma. This project also deals with determining the most efficient method for testing of water sample for qualitative as well as quantitative analysis using LIBS.
Identification and characterization of kidney stone remains one of the important analytical tasks in the medical field. Kidney stone is a common health complication throughout the world, which may cause severe pain, obstruction and infection of urinary tract, and can lead to complete renal damage. It commonly occurs in both sexes regardless of age. Kidney stones have different composition, although each stones have a major single characteristic component. A complete understanding of a sample properties and their function can only be feasible by utilizing elemental and molecular information simultaneously. Two laser based analytical techniques; Laser Induced Breakdown spectroscopy (LIBS) and Raman spectroscopy have been used to study different types of kidney stones from different patients. LIBS and Raman spectroscopy are highly complementary spectroscopic techniques, which provide elemental and molecular information of a sample. Q-switched Nd:YAG laser at 355 nm laser having energy 17mJ per pulse at 10 Hz repetition rate was used for getting LIBS spectra. Raman measurements were carried out using a home assembled micro-Raman spectrometer. Using the recorded Raman spectra of kidney stones, we were able to differentiate different kinds of kidney stones. LIBS spectra of the same stones are showing the evidence of C, Ca, H, and O and also suggest the presence of certain pigments.
We present an Opto-mechanical Door Locking System which is an optical system that combines a simple combination of a coherent light source (Laser) and a photodiode based sensor with focus toward security applications. The basic construct of the KEY comprises a Laser source in a cylindrical enclosure that slides perfectly into the LOCK. The Laser is pulsed at a fixed encrypted frequency unique to that locking system. Transistor-transistor logic (TTL) circuitry is used to achieve encryption. The casing of the key is designed in such a way that it will power the pulsing laser only when the key is inserted in the slot provided for it. The Lock includes a photo-sensor that will convert the detected light intensity to a corresponding electrical signal by decrypting the frequency. The lock also consists of a circuit with a feedback system that will carry the digital information regarding the encryption frequency code. The information received from the sensor is matched with the stored code; if found a perfect match, a signal will be sent to the servo to unlock the mechanical lock or to carry out any other operation. This technique can be incorporated in security systems for residences and safe houses, and can easily replace all conventional locks which formerly used fixed patterns to unlock. The major advantage of this proposed optomechanical system over conventional ones is that it no longer relies on a solid/imprinted pattern to perform its task and hence makes it almost impossible to tamper with.
Single-cell micro-Raman spectroscopy has been applied to explore cell differentiation in single, live, and malignant cells from two tumor cell lines. The spectra of differentiated cells exhibit substantial enhancement primarily in the intensities of protein peaks with concomitant decrease in intensities of O─P─O asymmetric stretching peaks in DNA/RNA. Principal component analyses show that the spectral score of differentiated cells tends to asymptotically approach that of spectra obtained from normal neural stem cells/progenitors. This lends credence to the notion that the observed spectral changes are specific to differentiation, since upon differentiation, malignant cells become less malignant and tend toward benignity.
Oral cancer together with pharyngeal cancer is the sixth most common malignancy reported worldwide and one with high mortality ratio among all malignancies [1]. Worldwide 450,000 new cases are estimated in 2014[2]. About 90% are a type of cancer called squamous cell carcinoma (SCC). SCC of the tongue is the most common oral malignancy accounting for approximately 40% of all oral carcinomas. One of the important factors for successful therapy of any malignancy is early diagnosis. Although considerable progress has been made in understanding the cellular and molecular mechanisms of tumorigenesis, lack of reliable diagnostic methods for early detection leading to delay in therapy is an important factor responsible for the increase in the mortality rate in various types of cancers. Spectroscopy techniques are extremely sensitive for the analysis of biochemical changes in cellular systems. These techniques can provide a valuable information on alterations that occur during the development of cancer. This is especially important in oral cancer, where "tumor detection is complicated by a tendency towards field cancerization, leading to multi-centric lesions" and "current techniques detect malignant change too late" [3], and "biopsies are not representative of the whole premalignant lesion". [4]
LIBS has been proven to be a robust elemental analysis tool attracting interest because of the wide applications. LIBS can be used for analysis of any type of samples i.e. environmental/physiological, regardless of its state of matter. Conventional spectroscopy techniques are good in analytical performance, but their sample preparation method is mostly destructive and time consuming. Also, almost all these methods are incapable of analysing multi elements simaltaneously. On the other hand, LIBS has many potential advantages such as simplicity in the experimental setup, less sample preparation, less destructive analysis of sample etc. In this paper, we report some of the biomedical applications of LIBS. From the experiments carried out on clinical samples (calcified tissues or teeth and gall stones) for trace elemental mapping and detection, it was found that LIBS is a robust tool for such applications. It is seen that the presence and relative concentrations of major elements (calcium, phosphorus and magnesium) in human calcified tissue (tooth) can be easily determined using LIBS technique. The importance of this study comes in anthropology where tooth and bone are main samples from which reliable data can be easily retrieved. Similarly, elemental composition of bile juice and gall stone collected from the same subject using LIBS was found to be similar. The results show interesting prospects for LIBS to study cholelithiasis (the presence of stones in the gall bladder, is a common disease of the gastrointestinal tract) better.
An ultra-sensitive hyphenated technique, high-performance liquid chromatography-laser-induced fluorescence detection protein profiling of saliva, is evaluated for early detection and diagnosis of oral premalignancy and malignancy. Calibration sets of protein profiles of unstimulated whole saliva are collected from clinically/pathologically normal, premalignant, and malignant subjects and used as standards. Three parameters—scores of factors, sum of squared residuals, and Mahalanobis distance—derived from principal component analysis of protein profiles of the standard calibration sets, and blind samples are used for “match/no-match” diagnosis of the blind samples. Analyses of the results show that the method is capable of differentiating normal, premalignant, and malignant conditions with the sensitivity and specificity of 79% and 78%, respectively. The technique provides a fast, highly objective (free from personal judgment and statistically defined), and noninvasive diagnostic method for screening and early detection of oral cancer.
The present work deals with the evaluation of a high-performance liquid chromatography laser-induced fluorescence (HPLC-LIF) technique developed in our laboratory for early detection of oral cancer from protein profiles of body fluids. The results show that protein profiles of serum samples from a given class of samples, say, normal, premalignant, or malignant, are statistically very close to each other, while profiles of members of any class are significantly different from other classes. The performance of the technique is evaluated by the use of sensitivity and specificity pairs, receiver operating characteristic (ROC) analysis, and Youden's Index. The technique uses protein profile differences in serum samples, registered by the HPLC-LIF technique. The study is carried out using serum samples from volunteers diagnosed as normal or premalignant clinically, and as malignant by histopathology. The specificities and sensitivities of the HPLC-LIF method at an ideal threshold (M-distance = 2) for normal, malignant, and premalignant classes are 100, 69.5, and 61.5%, and 86.5, 87.5, and 87.5% respectively.
The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded
using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF).
Serum protein profiles of the above three classes were tested for establishing the ability of HPLC-LIF protein profiling
technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite
successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions.
Protein profiles of tissue homogenates were recorded using HPLC separation and LIF detection method. The samples
were collected from volunteers with clinically normal or cervical cancer conditions. It is shown that the protein profile
can be classified as belonging to malignant or normal state by using hard and Fuzzy clustering methods. The study was
performed to test the utility of the HPLC-LIF protein profiling method for classification of tissue samples as well as to
establish a complementary method for histopathology for clinical diagnosis of the tissue as normal or malignant.
High performance liquid chromatography with high sensitivity laser-induced fluorescence detection is used to study the protein profiles of serum samples from healthy volunteers and cervical cancer subjects. The protein profiles are subjected to principal component analysis (PCA). PCA shows that the large number of chromatograms of a given class of serum samples—say normal/malignant—can be expressed in terms of a small number of factors (principal components). Three parameters—scores of the factors, squared residuals, and Mahalanobis distance—are derived from PCA. The parameters are observed to have a narrow range for protein profiles of standard calibration sets formed from groups of clinically confirmed normal/malignant classes. Limit tests using match/no match of the parameters of any test sample with parameters derived for the standard calibration sets give very good discrimination between malignant and normal samples with high sensitivity (~100%) and specificity (~94%).
The first studies of the propagation of ultrafast (<45 fs) pulses of intense infrared light through protein media reveal that supercontinuum (white light) generation is severely suppressed in the presence of the protein -amylase, a potential stress marker in human saliva. The continuum suppression capacity is attributed to the electron scavenging property of the protein.
HPLC combined with laser induced fluorescence provides a very sensitive method for the separation and identification
of the many proteins present in clinical samples. Protein profiles of clinical samples like Pap smear and tissue samples,
from subjects with cervical cancer and normal volunteers, were recorded using HPLC-LIF. The protein profiles were
analyzed by Principal Component Analysis (PCA). The profiles were characterized by parameters like scores of the
factors, sum of squared residuals, and Mahalanobis Distance, derived from PCA. Parameters of each sample were
compared with those of a standard set and Match/ No Match results were generated. Good discrimination between
normal and malignant samples was achieved with high sensitivity and specificity.
Propagation of ultrashort pulses of intense, infrared light through transparent medium gives rise to a visually spectacular phenomenon known as supercontinuum (white light) generation wherein the spectrum of transmitted light is very considerably broader than that of the incident light. We have studied the propagation of ultrafast (<45 fs) pulses of intense infrared light through biological media (water, and water doped with salivary proteins) which reveal that white light generation is severely suppressed in the presence of a major salivary protein, &agr;-amylase.
Proteomics based techniques are rapidly emerging as alternative techniques to conventional histo-pathological methods
for detection and diagnosis of cancers. Tumor markers are of considerable importance in the study of various cancers. A
study of various changes in the protein profile associated with breast cancer will facilitate a better understanding of the
various dynamic changes associated with the disease. In our study we have used High Performance Liquid
Chromatography coupled with highly sensitive Laser Induced Fluorescence for recording the protein profiles of breast
tissue homogenates. The protein profiles were recorded from pathologically certified normal as well as malignant breast
tissue samples. The recorded protein profiles were studied by using Principal Component Analysis. Good
discrimination of normal, benign and malignant samples was achieved in this pilot study.
Optical methods and proteomics investigations are becoming promising approaches for early detection of many diseases, which remain clinically silent for long periods. We have used efficient High Performance Liquid Chromatography (HPLC) separation combined with highly sensitive laser induced fluorescence detection of proteins present in clinical samples for diagnostic applications in cervical cancer. The protein profile and the fluorescence of individual proteins were simultaneously recorded using our HPLC-LIF system. Protein profiles (Chromatogram) of serum from normal male and female volunteers with and without tobacco habits, and malignant serum samples were studied. Protein profiles were also recorded for lysates of exfoliated cells collected from Pap smear of normal and cancer patients. The protein profile patterns were subjected to Principal component Analysis. Discrimination of normal and malignant samples were achieved with very high sensitivity and specificity.
New techniques for the early detection of cancer are fast emerging. This is essential for more effective diagnosis and control of the disease. We have used a High Performance Liquid Chromatography-Laser Induced Fluorescence (HPLCLIF) technique to record chromatograms of proteins in serum and ovarian tissue samples. The recorded chromatograms of normal, benign and malignant samples were analyzed using statistical (Principal Component Analysis) methods. It is shown that chromatograms of the samples can be classified into sets, and a model based on such a classification can be used to analyze protein profiles of test samples of serum and ovarian tissue for the detection of malignancies.
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