Raman spectroscopy assisted tear analysis: A label free, optical approach for noninvasive disease diagnostics.

In recent times, tear fluid analysis has garnered considerable attention in the field of biomarker-based diagnostics due to its noninvasive sample collection method. Tears encompass a reservoir of biomarkers that assist in diagnosing not only ocular disorders but also a diverse list of systemic diseases. This highlights the necessity for sensitive and dependable screening methods to employ tear fluid as a potential noninvasive diagnostic specimen in clinical environments. Considerable research has been conducted to investigate the potential of Raman spectroscopy-based investigations for tear analysis in various diagnostic applications. Raman Spectroscopy (RS) is a highly sensitive and label free spectroscopic technique which aids in investigating the molecular structure of samples by evaluating the vibrational frequencies of molecular bonds. Due to the distinct chemical compositions of different samples, it is possible to obtain a sample-specific spectral fingerprint. The distinctive spectral fingerprints obtained from Raman spectroscopy enable researchers to identify specific compounds or functional groups present in a sample, aiding in diverse biomedical applications. Its sensitivity to changes in molecular structure or environment provides invaluable insights into subtle alterations associated with various diseases. Thus, Raman Spectroscopy has the potential to assist in diagnosis and treatment as well as prognostic evaluation. Raman spectroscopy possesses several advantages, such as the non-destructive examination of samples, remarkable sensitivity to structural variations, minimal prerequisites for sample preparation, negligible interference from water, and the aptness for real-time investigation of tear samples. The purpose of this review is to highlight the potential of Raman spectroscopic technique in facilitating the clinical diagnosis of various ophthalmic and systemic disorders through non-invasive tear analysis. Additionally, the review delves into the advancements made in Raman spectroscopy with regards to paper-based sensing substrates and tear analysis methods integrated into contact lenses. Furthermore, the review also addresses the obstacles and future possibilities associated with implementing Raman spectroscopy as a routine diagnostic tool based on tear analysis in clinical settings.

Visualization and characterisation of microplastics in aquatic environment using a home-built micro-Raman spectroscopic set up.

Microplastics (MP) which are tiny plastic particles of sizes range from 1 µm (µm) to 5 mm (mm), have become a growing cause of concern due to their widespread presence in the environment and their potential impacts on ecosystems and human health. Marine organisms have the potential to consume microplastics, which could lead to physical injuries, blockages, or the transfer of harmful substances up the food chain. Humans may indirectly consume microplastics through contaminated seafood and water, although the complete scope of health risks is currently under investigation. An essential step in gaining a comprehensive understanding of microplastic pollution in waterbodies is the identification of microplastics, which is also crucial for further development of effective environmental regulations to address its adverse impacts. Majority of the researchers are accomplishing it globally using commercial platforms based on Raman spectroscopy. However, the development of indigenous Raman systems, which can enable microplastic identification, particularly in developing nations, is the need of the hour due to the outrageous cost of commercial platforms. In the current study, a custom-designed micro-Raman spectroscopy system was developed to detect and characterize microplastics from waterbodies. The developed system enabled visualization, size measurement and characterization of microplastics. Experimental parameters were fine-tuned, and a standardized Raman database was established for each type of plastic. This system exhibited high resolution which was capable of analysing microparticles of size up to 5 µm. Principal component analysis was carried out on the experimental Raman data, demonstrating good classification amongst different kinds of plastics. The performance of the developed system in analysing real samples was evaluated through experiments conducted on water samples obtained from the shore of Malpe Beach in Udupi district. The results revealed the presence of polyethylene and polyethylene terephthalate in the samples, along with the detection of pigments like copper phthalocyanine and indigo blue.

Micro-Raman spectroscopy study of blood samples from myocardial infarction patients.

Acute myocardial infarction (MI) is found to be a major causative factor for global mortality and morbidity. This situation demands necessity of developing efficient and rapid diagnostic tools to detect acute MI. Raman spectroscopy is a non-destructive optical diagnostic technique, which has high potential in probing biochemical changes in clinical samples during initiation and progress of diseases. In this work, blood was taken as the sample to examine inflammation in acute MI patients using Raman spectroscopy. Ratio of Raman peak intensities that corresponds to phenylalanine (1000 cm-1) and tyrosine (825 cm-1) can facilitate indirect information about tetrahydrobiopterin (BH4) availability, which can indicate inflammatory status in patients. This ratio obtained was higher for MI patients in comparison with control subjects. The decrease in phenylalanine and tyrosine ratio (Phe-Tyr ratio) is attributed to the prognosis of standard of care (medications like antiplatelets including aspirin, statin and revascularisation) leading to inflammation reduction. Phe-Tyr ratio estimated from the Raman spectra of blood can be exploited as a reliable method to probe inflammation due to MI. The method is highly objective, require only microliters of sample and minimal sample preparation, signifying its clinical utility.

A micro-Raman spectroscopy study of inflammatory condition of human cervix: Probing of tissues and blood plasma samples.

The present study explores the application of the micro-Raman spectroscopy technique to discriminate normal and cervicitis condition from cervical malignancy by analyzing the Raman signatures of tissues and plasma samples of the same subjects. The Raman peaks from tissue samples at 1026 cm-1,1298 cm-1 and 1243 cm-1 are attributed to glycogen, fatty acids and collagen and are found to be reliable signatures capable of identifying cervicitis and normal condition from cervical cancer. The Raman signatures from plasma samples belonging to carbohydrates (578 cm-1), lipids (1059 cm-1) and nucleic acids (1077 cm-1,1341 cm-1 and 1357 cm-1) are quite useful to classify various pathological conditions of cervix at par with tissue based diagnosis. The PCA-SVM based classification of the spectral data indicates the potential of Raman spectroscopy based liquid biopsy to rule out false diagnosis of cervicitis as cervical malignancy.

Photonics of human saliva: potential optical methods for the screening of abnormal health conditions and infections.

Human saliva can be treated as a pool of biological markers able to reflect on the state of personal health. Recent years have witnessed an increase in the use of optical devices for the analysis of body fluids. Several groups have carried out studies investigating the potential of saliva as a non-invasive and reliable clinical specimen for use in medical diagnostics. This brief review aims to highlight the optical technologies, mainly surface plasmon resonance (SPR), Raman, and Fourier transform infrared (FTIR) spectroscopy, which are being used for the probing of saliva for diverse biomedical applications. Advances in bio photonics offer the promise of unambiguous, objective and fast detection of abnormal health conditions and viral infections (such as COVID-19) from the analysis of saliva.

Single cell spectroscopy of red blood cells in intravenous crystalloid fluids.

Crystalloid fluids, a subset of intravenous (IV) fluid solutions are commonly used in clinical settings. The influence of these fluids on the functions of blood components are least explored. Raman spectroscopy combined with optical trapping has been widely used to evaluate the impact of external stress agents on red blood cells. The present study investigates the impact of commonly used crystalloid fluids on red blood cells in comparison with that of blood plasma using Raman Tweezers spectroscopy. The red blood cells suspended in crystalloid fluids undergo deoxygenation readily than that in blood plasma. In addition, cells in blood plasma were able to withstand laser induced deoxygenation comparatively better than that in crystalloid fluids at higher laser powers. Principle component analysis of the Raman spectral data has clearly demonstrated the discrimination of cells in plasma with that of crystalloid fluids demonstrating the effect of external induced stress on RBCs.

Human red blood cell behaviour in hydroxyethyl starch: probed by single cell spectroscopy.

Hydroxyethyl starch (HES) is a commonly used intravenous fluid in hospital settings. The merits and demerits of its application is still a debatable topic. Investigating the interaction of external agents like intravenous fluids with blood cells is of great significance in clinical environments. Micro-Raman spectroscopy combined with an optical tweezers technique has been utilized for conducting systematic investigations of single live red blood cells (RBCs) under the influence of external stress agents. The present work deals with a detailed biophysical study on the response of human live red blood cells in hydroxyethyl starch using optical techniques. Morphological changes in red blood cells were monitored using quantitate phase imaging techniques. Micro-Raman studies suggest that there is a significant reduction in the oxy-haemoglobin level in red blood cells suspended in HES. The spectra recorded by using different probe laser powers has shown that the cells are more vulnerable in HES under the influence of externally induced stress than in blood plasma. In addition, the spectral results support the possibility of heme aggregation and membrane damage for red blood cells in HES under externally induced stress. Principle component analysis performed on the Raman spectra were able to effectively discriminate between red blood cells in HES and in blood plasma. The use of Raman tweezers can be highly beneficial in elucidating biochemical alterations happening in live, human red blood cell.

Normal saline-induced deoxygenation of red blood cells probed by optical tweezers combined with the micro-Raman technique.

The use of normal saline for washing red blood cells and treating critically ill patients is a regular medical practice in hospital settings. An optical tweezer in combination with Raman spectroscopy is an analytical tool employed for the investigation of single cell dynamics, thus providing molecular fingerprint of the cell by optically trapping the cell at a laser focus. In this study, the impact of normal saline on individual human red blood cell was compared with that of blood plasma using Raman tweezers spectroscopy. Major spectral variations in the marker frequencies at 1209 cm-1, 1222 cm-1, 1544 cm-1, and 1561 cm-1 of the Raman spectrum of the treated cells imply that the transition of hemoglobin to the deoxygenated state occurs when 0.9% normal saline is used. This may result in serious implications in blood transfusion. The results obtained from the principal component analysis also displayed clear differentiation among the red blood cells diluted in normal saline and those diluted in plasma. In future studies, efforts will be made to correlate the deoxygenation status of red blood cells with various human disorders.

Laser Raman tweezer spectroscopy to explore the bisphenol A-induced changes in human erythrocytes.

The dermal penetration of bisphenol-A (BPA) from thermal papers into the human skin is a matter of major health concern due to its extensive use in developing countries like India, one of its largest users in the world. Bisphenol A is widely used in the manufacture of many consumer goods like polycarbonate water bottles, baby bottles, food containers, home appliances, thermal papers used in billing and tickets, the inner lining of food cans, etc. BPA can be easily adsorbed into the blood rapidly. The integration of optical tweezers with Raman spectroscopic techniques has realized avenues for interpreting single cell investigations. In the present work, the impact of BPA from thermal papers on individual human erythrocytes (red blood cells) has been investigated using micro-Raman spectroscopy. Significant intensity variations were noticed for hemoglobin oxygenation markers in the Raman spectra of red blood cells (RBCs). Raman spectral variations supporting RBC hemoglobin depletion were also found in the presence of BPA. Evident morphological changes are also observed in RBCs due to BPA in vitro exposures, which ultimately lead to cell bursting at higher concentrations.