Reliable and easy-to-use SERS spectroscopy probe using a tapered opto-fluidic photonic crystal fiber.

Surface enhanced Raman spectroscopy (SERS) is one of the most sensitive biosensing techniques that offers label free detection for a variety of applications. Generally, SERS spectroscopy is performed on nano-functionalized planar substrates with plasmonic structures or colloidal nanoparticles. Recently, photonic crystal fibers (PCFs) have gained great interest for SERS based bio sensing applications due to the immense advantages such as improved sensitivity, flexibility and remote sensing capability that it offers compared to the planar substrates. However, the use of PCF based biosensors demand the alignment of it under a microscope, which can affect the reliability of SERS measurements and could be restrictive for practical end use applications. Herein, we aim to develop a tapered suspended core PCF fiber (Tapered-SuC-PCF) that represents an improvement in coupling efficiency and measurement reliability as well as it opens the way to the development of an easy-to-use bio-sensing probes with a plug and play option with conventional Raman spectrometers. We have fabricated several samples of the optimized tapered-SuC-PCF and demonstrated its superior SERS performance compared to standard SuC-PCF fibers with 2 µm core diameter. An excellent SERS measurement reliability is demonstrated using such a fiber in a plug and play type system demonstrating its versatility for practical end use applications.

Optimization and performance analysis of SERS-active suspended core photonic crystal fibers.

Recently, surface enhanced Raman spectroscopy (SERS)-active photonic crystal fiber (PCFs) probes have gained great interest for biosensing applications due to the tremendous advantages it has over the conventional planar substrate based SERS measurements, with improvements on the detection sensitivity and reliability in measurements. So far, two main approaches were employed to get the analyte molecule in the vicinity of nanoparticles (NPs) inside PCFs in order to achieve the SERS effect. In the first case, analyte and NPs are pre-mixed and injected inside the holes of the PCF prior to the measurement. In the second approach, controlled anchoring of the NPs inside the inner walls of the PCF was achieved prior to the incorporation of the analyte. Although many studies have been conducted using one configuration or the other, no clear trend is emerging on which one would be the best suited for optimizing the biosensing properties offered by SERS active-PCF. In this paper, we investigate the performances of both configurations along with their interplays with the core size of the PCF probe. We have fabricated several samples of a standard PCF design with different core sizes, and SERS measurements of a standard Raman-active molecule are realized in the same conditions for enabling direct comparisons of the SERS intensity and measurement reliabilities between each configuration, yielding clear directions on the optimization of the SERS-active PCF probe. We envision that this study will pave the way for next-generation clinical biosensors for body fluid analysis, as it exhibits high sensitivity and excellent reliability.

Optofluidic photonic crystal fiber platform for sensitive and reliable fluorescence based biosensing.

Biosensing plays a pivotal role in various scientific domains, offering significant contributions to medical diagnostics, environmental monitoring, and biotechnology. Fluorescence biosensing relies on the fluorescence emission from labelled biomolecules to enable sensitive and selective identification and quantification of specific biological targets in various samples. Photonic crystal fibers (PCFs) have led to the development of optofluidic fibers enabling efficient light-liquid interaction within small liquid volume. Herein, we present the development of a user-friendly optofluidic-fiber platform with simple hardware requirements for sensitive and reliable fluorescence biosensing with high measurement repeatability. We demonstrate a sensitivity improvement of the fluorescence emission up to 17 times compared to standard cuvette measurement, with a limit of detection of Cy5 fluorophore as low as 100 pM. The improvement in measurement repeatability is exploited for detecting haptoglobin protein, a relevant biomarker to diagnose several diseases, by using commercially available Cy5 labelled antibodies. The study aims to showcase an optofluidic platform leveraging the benefits provided by optofluidic fibers, which encompass easy light injection, robustness, and high sensitivity.

Complete characterization of RNA biomarker fingerprints using a multi-modal ATR-FTIR and SERS approach for label-free early breast cancer diagnosis.

Breast cancer is a prevalent form of cancer worldwide, and the current standard screening method, mammography, often requires invasive biopsy procedures for further assessment. Recent research has explored microRNAs (miRNAs) in circulating blood as potential biomarkers for early breast cancer diagnosis. In this study, we employed a multi-modal spectroscopy approach, combining attenuated total reflection Fourier transform infrared (ATR-FTIR) and surface-enhanced Raman scattering (SERS) to comprehensively characterize the full-spectrum fingerprints of RNA biomarkers in the blood serum of breast cancer patients. The sensitivity of conventional FTIR and Raman spectroscopy was enhanced by ATR-FTIR and SERS through the utilization of a diamond ATR crystal and silver-coated silicon nanopillars, respectively. Moreover, a wider measurement wavelength range was achieved with the multi-modal approach than with a single spectroscopic method alone. We have shown the results on 91 clinical samples, which comprised 44 malignant and 47 benign cases. Principal component analysis (PCA) was performed on the ATR-FTIR, SERS, and multi-modal data. From the peak analysis, we gained insights into biomolecular absorption and scattering-related features, which aid in the differentiation of malignant and benign samples. Applying 32 machine learning algorithms to the PCA results, we identified key molecular fingerprints and demonstrated that the multi-modal approach outperforms individual techniques, achieving higher average validation accuracy (95.1%), blind test accuracy (91.6%), specificity (94.7%), sensitivity (95.5%), and F-score (94.8%). The support vector machine (SVM) model showed the best area under the curve (AUC) characterization value of 0.9979, indicating excellent performance. These findings highlight the potential of the multi-modal spectroscopy approach as an accurate, reliable, and rapid method for distinguishing between malignant and benign breast tumors in women. Such a label-free approach holds promise for improving early breast cancer diagnosis and patient outcomes.

Structural and functional imaging of psoriasis for severity assessment and quantitative monitoring of treatment response using high-resolution optoacoustic imaging.

Psoriasis is a chronic inflammatory skin disease, characterized by thick scaly plaques. It imposes a notable disease burden with varying levels of severity affecting the quality of life significantly. Current disease severity assessment relies on semi-objective visual inspection based on the Psoriasis Area and Severity index (PASI) score that might not be sensitive to sub-clinical changes. Histology of psoriasis skin lesions necessitate invasive skin biopsies. This indicates an unmet need for a non-invasive, objective and quantitative approach to assess disease severity serially. Herein, we employ multispectral Raster-Scanning Optoacoustic Mesoscopy (ms-RSOM) derived structural and microvascular functional imaging metrics to examine the lesional and non-lesional skin in psoriasis subjects across different severities and also evaluate the treatment outcome in a subject with topical steroids and biologics, such as adalimumab. ms-RSOM derived structural metrics like epidermal thickness and total blood volume (TBV) and microvascular functional information such as oxygen saturation (sO2) are evaluated by spectrally resolving the endogenous chromophores like melanin, oxy-, and deoxy-hemoglobin. Initial findings reveal an elevated sO2 and TBV with severity in lesional and non-lesional psoriasis skin, thus representing increasing inflammation. An increase in epidermal thickness is also noted with the degree of severity, corresponding to the inflammation and increased abnormal cell growth. As a marker to evaluate the treatment response, we observed a decrease in epidermal thickness, sO2, and TBV in a psoriasis patient post-treatment, which is consistent with the decrease in the PASI score from 4.1 to 1.9. We envision that ms-RSOM has a huge potential to be translated into routine clinical setting for the diagnosis of severity and assessment of treatment monitoring in psoriasis subjects.

Diffuse reflectance spectroscopy and imaging for non-invasive objective assessment of genitourinary syndrome of menopause: a pilot study.

The genitourinary symptom of menopause (GSM) affects up to 65% of women, resulting in symptoms such as vulvovaginal dryness, discomfort, and dysuria, which significantly impacts quality of life. The current assessment methods rely on subjective questionnaires that can be influenced by individual differences, as well as invasive measurements that are time-consuming and not easily accessible. In this study, we explore the potential of a non-invasive and objective assessment tool called diffuse reflectance spectroscopy and imaging (DRSI) to evaluate tissue chromophores, including water, lipid, oxyhemoglobin, and deoxyhemoglobin. These measurements provide information about moisture content, lipid levels, oxygen saturation, and blood fraction, which can serve as surrogate markers for genital estrogen levels. Our findings reveal distinct differences in these chromophores among pre, peri, and postmenopausal subjects. By using lipid and blood fraction tissue chromophores in a K-Nearest Neighbour classifier model, we achieved a prediction accuracy of 65% compared to vaginal maturation index (VMI) that is clinically used to assess estrogen-related hormonal changes. When age was included as the third feature, the accuracy increased to 78%. We believe that by refining the study protocol and configuring the fiber probe to examine tissue chromophores both in the superficial vulva skin for epidermal water content and the deeper layers, DRSI has the potential to provide objective diagnosis and aid in monitoring the treatment outcome of GSM.

Tunable Tamm plasmon cavity as a scalable biosensing platform for surface enhanced resonance Raman spectroscopy.

Surface enhanced Resonance Raman spectroscopy (SERRS) is a powerful technique for enhancing Raman spectra by matching the laser excitation wavelength with the plasmonic resonance and the absorption peak of biomolecules. Here, we propose a tunable Tamm plasmon polariton (TPP) cavity based on a metal on distributed Bragg reflector (DBR) as a scalable sensing platform for SERRS. We develop a gold film-coated ultralow-loss phase change material (Sb2S3) based DBR, which exhibits continuously tunable TPP resonances in the optical wavelengths. We demonstrate SERRS by matching the TPP resonance with the absorption peak of the chromophore molecule at 785 nm wavelength. We use this platform to detect cardiac Troponin I protein (cTnI), a biomarker for early diagnosis of cardiovascular disease, achieving a detection limit of 380 fM. This scalable substrate shows great promise as a next-generation tunable biosensing platform for detecting disease biomarkers in body fluids for routine real-time clinical diagnosis.

Miniaturized Vis-NIR handheld spectrometer for non-invasive pigment quantification in agritech applications.

Advanced precision agriculture requires the objective measurement of the structural and functional properties of plants. Biochemical profiles in leaves can differ depending on plant growing conditions. By quantitatively detecting these changes, farm production processes can be optimized to achieve high-yield, high-quality, and nutrient dense agricultural products. To enable the rapid and non-destructive detection on site, this study demonstrates the development of a new custom-designed portable handheld Vis-NIR spectrometer that collects leaf reflectance spectra, wirelessly transfers the spectral data through Bluetooth, and provides both raw spectral data and processed information. The spectrometer has two preprogramed methods: anthocyanin and chlorophyll quantification. Anthocyanin content of red and green lettuce estimated with the new spectrometer showed an excellent correlation coefficient of 0.84 with those determined by a destructive gold standard biochemical method. The differences in chlorophyll content were measured using leaf senescence as a case study. Chlorophyll Index calculated with the handheld spectrometer gradually decreased with leaf age as chlorophyll degrades during the process of senescence. The estimated chlorophyll values were highly correlated with those obtained from a commercial fluorescence-based chlorophyll meter with a correlation coefficient of 0.77. The developed portable handheld Vis-NIR spectrometer could be a simple, cost-effective, and easy to operate tool that can be used to non-invasively monitor plant pigment and nutrient content efficiently.

Non-invasive biochemical analysis and comparison of atopic dermatitis and psoriasis skin using handheld confocal Raman spectroscopy.

A handheld non-invasive confocal Raman system (CRS) was used to evaluate the differences in skin biochemicals between atopic dermatitis (AD) and psoriasis, which are inflammatory skin conditions. Raman spectral measurements in the fingerprint and high wavenumber region were acquired using a portable in-house CRS system with excitation lasers operating at 671 and 785 nm. It was deduced that relative amount of water decreases in the following sequence of skin: healthy, psoriasis and AD. Moreover, differential trends were observed for the subclasses of ceramides such that ceramide 3 is lower in the lesional AD and psoriasis skin as compared to healthy, while ceramide 2 showed a contrasting trend of decrease in lesional AD and increase in lesional psoriasis as opposed to healthy skin. Amount of cholesterol was significantly higher in lesional psoriasis as compared to lesional AD and healthy skin. These differences can aid in an objective classification of the skin conditions and in the formulation of new disease-specific topical treatments.

A pilot study on non-invasive in situ detection of phytochemicals and plant endogenous status using fiber optic infrared spectroscopy.

Traditional methods for assessing plant health often lack the necessary attributes for continuous and non-destructive monitoring. In this pilot study, we present a novel technique utilizing a customized fiber optic probe based on attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) with a contact force control unit for non-invasive and continuous plant health monitoring. We also developed a normalized difference mid-infrared reflectance index through statistical analysis of spectral features, enabling differentiation of drought and age conditions in plants. Our research aims to characterize phytochemicals and plant endogenous status optically, addressing the need for improved analytical measurement methods for in situ plant health assessment. The probe configuration was optimized with a triple-loop tip and a 3 N contact force, allowing sensitive measurements while minimizing leaf damage. By combining polycrystalline and chalcogenide fiber probes, a comprehensive wavenumber range analysis (4000-900 cm-1) was achieved. Results revealed significant variations in phytochemical composition among plant species, for example, red spinach with the highest polyphenolic content and green kale with the highest lignin content. Petioles displayed higher lignin and cellulose absorbance values compared to veins. The technique effectively monitored drought stress on potted green bok choy plants in situ, facilitating the quantification of changes in water content, antioxidant activity, lignin, and cellulose levels. This research represents the first demonstration of the potential of fiber optic ATR-FTIR probes for non-invasive and rapid plant health measurements, providing insights into plant health and advancements in quantitative monitoring for indoor farming practices, bioanalytical chemistry, and environmental sciences.

Nanosphere templated metallic grating assisted enhanced fluorescence.

In this paper, enhanced fluorescence from a silver film coated nanosphere templated grating is presented. Initially, numerical simulation was performed to determine the plasmon resonance wavelength by varying the thickness of the silver film on top of a monolayer of 400 nm nanospheres. The simulation results are verified experimentally and tested for enhancing fluorescence from fluorescein isothiocyanate whose excitation wavelength closely matches with the plasmon resonance wavelength of the substrate with 100 nm silver film over nanosphere. The 12 times enhancement in the intensity is attributed to the local field enhancement in addition to the excitation of surface plasmon polaritons along the surface.

Machine Learning Assisted Real-Time Label-Free SERS Diagnoses of Malignant Pleural Effusion due to Lung Cancer.

More than half of all pleural effusions are due to malignancy of which lung cancer is the main cause. Pleural effusions can complicate the course of pneumonia, pulmonary tuberculosis, or underlying systemic disease. We explore the application of label-free surface-enhanced Raman spectroscopy (SERS) as a point of care (POC) diagnostic tool to identify if pleural effusions are due to lung cancer or to other causes (controls). Lung cancer samples showed specific SERS spectral signatures such as the position and intensity of the Raman band in different wave number region using a novel silver coated silicon nanopillar (SCSNP) as a SERS substrate. We report a classification accuracy of 85% along with a sensitivity and specificity of 87% and 83%, respectively, for the detection of lung cancer over control pleural fluid samples with a receiver operating characteristics (ROC) area under curve value of 0.93 using a PLS-DA binary classifier to distinguish between lung cancer over control subjects. We have also evaluated discriminative wavenumber bands responsible for the distinction between the two classes with the help of a variable importance in projection (VIP) score. We found that our label-free SERS platform was able to distinguish lung cancer from pleural effusions due to other causes (controls) with higher diagnostic accuracy.

Handheld Multifunctional Fluorescence Imager for Non-invasive Plant Phenotyping.

Fluorescence imaging has shown great potential in non-invasive plant monitoring and analysis. However, current systems have several limitations, such as bulky size, high cost, contact measurement, and lack of multifunctionality, which may hinder its applications in a wide range of settings including indoor vertical farming. Herein, we developed a compact handheld fluorescence imager enabling multipurpose plant phenotyping, such as continuous photosynthetic activity monitoring and non-destructive anthocyanin quantification. The compact imager comprises of pulse-amplitude-modulated multi-color light emitting diodes (LEDs), optimized light illumination and collection, dedicated driver circuit board, miniaturized charge-coupled device camera, and associated image analytics. Experiments conducted in drought stressed lettuce proved that the novel imager could quantitatively evaluate the plant stress by the non-invasive measurement of photosynthetic activity efficiency. Moreover, a non-invasive and fast quantification of anthocyanins in green and red Batavia lettuce leaves had excellent correlation (>84%) with conventional destructive biochemical analysis. Preliminary experimental results emphasize the high throughput monitoring capability and multifunctionality of our novel handheld fluorescence imager, indicating its tremendous potential in modern agriculture.

Multispectral raster-scanning optoacoustic mesoscopy differentiate lesional from non-lesional atopic dermatitis skin using structural and functional imaging markers.

Atopic dermatitis (AD) is a chronic and pruritic skin inflammatory disease causing a significant burden to health care management and patient's quality of life. Seemingly healthy skin or non-lesional sites on AD patients still presents skin barrier defects and immune response, which can develop to AD at a later stage. To investigate further the balance between the epidermal barrier impairment and intrinsic immune dysregulation in AD, we exploited multispectral Raster-Scanning Optoacoustic Mesoscopy (ms-RSOM) to image lesional and non-lesional skin areas on AD patients of different severities non-invasively to elucidate their structural features and functional information. Herein, we demonstrate the objective assessment of AD severity using relative changes in oxygen saturation (δsO2) levels in microvasculature along with other structural parameters such as relative changes in epidermis thickness (δET) and total blood volume (δTBV) between the lesional and non-lesional areas of the skin. We could observe an increasing trend for δsO2 and δTBV, which correlated well with the subjective clinical Scoring Atopic Dermatitis (SCORAD) for evaluating the severity. Notably, δET showed a decreasing trend with AD severity, indicating that the difference in epidermal thickness between lesional and non-lesional area of the skin decreases with AD severity. Our results also correlated well with conventional metrics such as trans-epidermal water loss (TEWL) and erythrosine sedimentation rate (ESR). We quantified the δsO2 and δET changes to objectively evaluate the treatment response before and four months after treatment using topical steroids and cyclosporine in one severe AD patient. We observed reduced δsO2 and δET post treatment. We envision that in future, functional and structural imaging metrics derived from ms-RSOM can be translated as objective markers to assess and stratify the severity of AD and understand the function of skin barrier dysfunctions and immune dysregulation. It could also be employed to monitor the treatment response of AD in regular clinical settings.

Towards a point-of-care SERS sensor for biomedical and agri-food analysis applications: a review of recent advancements.

The growing demand for reliable and robust methodology in bio-chemical sensing calls for the continuous advancement of sensor technologies. Over the last two decades, surface-enhanced Raman spectroscopy (SERS) has emerged as one of the most promising analytical techniques for sensitive and trace analysis or detection in biomedical and agri-food applications. SERS overcomes the inherent sensitivity limitation associated with Raman spectroscopy, which provides vibrational "fingerprint" spectra of molecules that makes it unique and versatile among other spectroscopy techniques. This paper comprehensively reviews the recent advancements of SERS for biomedical, food and agricultural applications over the last 6 years, and we envision that, in the near future, some of these platforms have the potential to be translated as a point-of-care and rapid sensor for real-life end-user applications. The merits and limitations of various SERS sensor designs are analysed and discussed based on critical features such as sensitivity, specificity, usability, repeatability and reproducibility. We conclude by highlighting the opportunities and challenges in the field while stressing the technological gaps to be addressed in realizing commercially viable point-of-care SERS sensors for practical biomedical and agri-food technological applications.

A Portable Ultrawideband Confocal Raman Spectroscopy System with a Handheld Probe for Skin Studies.

Confocal Raman spectroscopy (CRS) has shown potential in non-invasive skin analysis. However, current CRS systems have various limitations including a narrow detection band, large size, non-flexibility, slowness, and complexity, which hinder their clinical applications. Herein, we developed a portable ultrawideband CRS system with a fiber-based handheld probe to acquire the Raman spectra in both fingerprint and high wavenumber regions in a fast and quasi-simultaneous way. Dual-wavelength excitation with a dual-passband laser cleaning filter and high-speed fiber array multiplexer was adopted instead of a specialized grating and detector to achieve instant switching between the detection regions and improve system robustness. Preliminary in vivo results demonstrated its depth profiling capability in an ultrawide detection range for stratum corneum thickness, natural moisturizing factor, and water content quantification, indicating its great potential in a wide range of clinical and cosmeceutical applications.

Microvascular imaging and monitoring of hemodynamic changes in the skin during arterial-venous occlusion using multispectral raster-scanning optoacoustic mesoscopy.

The ability to monitor oxygen delivery in microvasculature plays a vital role in measuring the viability of skin tissue and the probability of recovery. Using currently available clinical imaging tools, it is difficult to observe non-invasive hemodynamic regulation in the peripheral vessels. Here we propose the use of a novel multispectral raster-scanning optoacoustic mesoscopy (RSOM) system for noninvasive clinical monitoring of hemodynamic changes in the skin microvasculature's oxy- (HbO2) and deoxy-hemoglobin (Hb), total hemoglobin (HbT) and oxygen saturation (rsO2). High resolution images of hemoglobin distribution in the skin microvasculature from six healthy volunteers during venous and arterial occlusion, simulating systemic vascular diseases are presented. During venous occlusion, Hb and HbO2 optoacoustic signals showed an increasing trend with time, followed by a drop in the values after cuff deflation. During arterial occlusion, an increase in Hb value and decrease in HbO2 values was observed, followed by a drop in Hb and jump in HbO2 values after the cuff deflation. A decrease in rsO2 values during both venous and arterial occlusion was observed with an increase in value after occlusion release. Using this proof of concept study, hereby we propose multispectral RSOM as a novel tool to measure high resolution hemodynamic changes in microvasculature for investigating systemic vascular diseases on peripheral tissues and also for monitoring inflammatory skin diseases, and its therapeutic interventions.

Novel Cellulose Fibre-Based Flexible Plasmonic Membrane for Point-of-Care SERS Biomarker Detection in Chronic Wound Healing.

BACKGROUND: Wound management is stretching the limits of health systems globally, challenging clinicians to evaluate the effectiveness of their treatments and deliver appropriate care to their patients. Visual inspection and manual measurement of wound size are subjective, often inaccurate and inconsistent. Growth factors, such as pro-inflammatory cytokines and proteases, play important roles in cutaneous wound healing. However, little is known about the point-of-care monitoring of the changes in such markers during the healing process. Here, we explore the capability of surface-enhanced Raman spectroscopy (SERS) as a viable point-of-care platform to monitor the changes of these surrogate indicators of healing status in chronic wounds. METHODS: We developed a biofunctionalized flexible, cost-effective, scalable and easy-to-fabricate plasmonic SERS substrate using cellulose fibre (CF), which is used for sensing of wound markers based on a modified immunoassay method. RESULTS: We evaluated and selected the reliable silver nano-island thickness that will be sputtered onto the CF-based substrate for the highest SERS enhancement. Using this biofunctionalized SERS substrate, we detected varying concentrations of MMP-9 (10-5000 ng/mL) and TNF-α (5-100 ng/mL) proteins to model the wound exudates. This SERS detection method demonstrates a linear response within biologically relevant concentrations, ranging from 10 to 500 ng/mL for MMP-9 and 5 to 25 ng/mL for TNF-α for these surrogate indicators. CONCLUSION: Our SERS sensing platform achieved detection limits in the µM to sub-nM range and displayed high sensitivity and selectivity. This could result in a cheap, point-of-care device that provides a non-invasive measure of cutaneous wound healing in real time. We envision that these flexible substrates after activation may be incorporated into wound dressings in future for routine monitoring of wound healing status.

Model learning analysis of 3D optoacoustic mesoscopy images for the classification of atopic dermatitis.

Atopic dermatitis (AD) is a skin inflammatory disease affecting 10% of the population worldwide. Raster-scanning optoacoustic mesoscopy (RSOM) has recently shown promise in dermatological imaging. We conducted a comprehensive analysis using three machine-learning models, random forest (RF), support vector machine (SVM), and convolutional neural network (CNN) for classifying healthy versus AD conditions, and sub-classifying different AD severities using RSOM images and clinical information. CNN model successfully differentiates healthy from AD patients with 97% accuracy. With limited data, RF achieved 65% accuracy in sub-classifying AD patients into mild versus moderate-severe cases. Identification of disease severities is vital in managing AD treatment.

Development of highly reliable SERS-active photonic crystal fiber probe and its application in the detection of ovarian cancer biomarker in cyst fluid.

Conventionally Surface-enhanced Raman spectroscopy (SERS) is realized by adsorbing analytes onto nano-roughened planar substrate coated with noble metals (silver or gold) or their colloidal nanoparticles (NPs). Nanoscale irregularities in such substrates/NPs could lead to SERS sensors with poor reproducibility and repeatability. Herein, we demonstrate a suspended core photonic crystal fiber (PCF) based SERS sensor with extremely high reproducibility and repeatability in measurement with a relative SD of only 1.5% and 4.6%, respectively, which makes it more reliable than any existing SERS sensor platforms. In addition, our platform could improve the detection sensitivity owing to the increased interaction area between the guided light and the analyte, which is incorporated into the holes that runs along the length of the PCF. Numerical calculation established the significance of the interplay between light coupling efficiency and evanescent field distribution, which could eventually determine the sensitivity and reliability of the developed SERS active-PCF sensor. As a proof of concept, using this sensor, we demonstrated the detection of haptoglobin, a biomarker for ovarian cancer, contained within the ovarian cyst fluid, which facilitated in differentiating the stages of cancer. We envision that with necessary refinements, this platform could potentially be translated as a next-generation highly sensitive SERS-active opto-fluidic biopsy needle for the detection of biomarkers in body fluids.