Biosensors

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13 pages, 2693 KiB

Open AccessArticle

Waveguiding and SERS Simplified Raman Spectroscopy on Biological Samples

by Immanuel Valpapuram, Patrizio Candeloro, Maria Laura Coluccio, Elvira Immacolata Parrotta, Andrea Giugni, Gobind Das, Gianni Cuda, Enzo Di Fabrizio and Gerardo Perozziello

Biosensors 2019, 9(1), 37; https://doi.org/10.3390/bios9010037 - 3 Mar 2019

Cited by 12 | Viewed by 7153

Abstract

Biomarkers detection at an ultra-low concentration in biofluids (blood, serum, saliva, etc.) is a key point for the early diagnosis success and the development of personalized therapies. However, it remains a challenge due to limiting factors like (i) the complexity of [...] Read more.

Biomarkers detection at an ultra-low concentration in biofluids (blood, serum, saliva, etc.) is a key point for the early diagnosis success and the development of personalized therapies. However, it remains a challenge due to limiting factors like (i) the complexity of analyzed media, and (ii) the aspecificity detection and the poor sensitivity of the conventional methods. In addition, several applications require the integration of the primary sensors with other devices (microfluidic devices, capillaries, flasks, vials, etc.) where transducing the signal might be difficult, reducing performances and applicability. In the present work, we demonstrate a new class of optical biosensor we have developed integrating an optical waveguide (OWG) with specific plasmonic surfaces. Exploiting the plasmonic resonance, the devices give consistent results in surface enhanced Raman spectroscopy (SERS) for continuous and label-free detection of biological compounds. The OWG allows driving optical signals in the proximity of SERS surfaces (detection area) overcoming spatial constraints, in order to reach places previously optically inaccessible. A rutile prism couples the remote laser source to the OWG, while a Raman spectrometer collects the SERS far field scattering. The present biosensors were implemented by a simple fabrication process, which includes photolithography and nanofabrication. By using such devices, it was possible to detect cell metabolites like Phenylalanine (Phe), Adenosine 5-triphosphate sodium hydrate (ATP), Sodium Lactate, Human Interleukin 6 (IL6), and relate them to possible metabolic pathway variation. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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19 pages, 4717 KiB

Open AccessArticle

Surface Enhanced Raman Spectroscopy of Lactoferrin Adsorbed on Silvered Porous Silicon Covered with Graphene

by Sergey Zavatski, Nadia Khinevich, Kseniya Girel, Sergey Redko, Nikolai Kovalchuk, Ivan Komissarov, Vladimir Lukashevich, Igor Semak, Kahramon Mamatkulov, Maria Vorobyeva, Grigory Arzumanyan and Hanna Bandarenka

Biosensors 2019, 9(1), 34; https://doi.org/10.3390/bios9010034 - 28 Feb 2019

Cited by 26 | Viewed by 7296

Abstract

We registered surface enhanced Raman scattering (SERS) spectra of the human lactoferrin molecules adsorbed on a silvered porous silicon (por-Si) from 10⁻⁶–10⁻¹⁸ M solutions. It was found that the por-Si template causes a negative surface potential of [...] Read more.

We registered surface enhanced Raman scattering (SERS) spectra of the human lactoferrin molecules adsorbed on a silvered porous silicon (por-Si) from 10⁻⁶–10⁻¹⁸ M solutions. It was found that the por-Si template causes a negative surface potential of silver particles and their chemical resistivity to oxidation. These properties provided to attract positively charged lactoferrin molecules and prevent their interaction with metallic particles upon 473 nm laser excitation. The SERS spectra of lactoferrin adsorbed from 10⁻⁶ M solution were rather weak but a decrease of the concentration to 10⁻¹⁰ M led to an enormous growth of the SERS signal. This effect took place as oligomers of lactoferrin were broken down to monomeric units while its concentration was reduced. Oligomers are too large for a uniform overlap with electromagnetic field from silver particles. They cannot provide an intensive SERS signal from the top part of the molecules in contrast to monomers that can be completely covered by the electromagnetic field. The SERS spectra of lactoferrin at the 10⁻¹⁴ and 10⁻¹⁶ M concentrations were less intensive and started to change due to increasing contribution from the laser burned molecules. To prevent overheating the analyte molecules on the silvered por-Si were protected with graphene, which allowed the detection of lactoferrin adsorbed from the 10⁻¹⁸ M solution. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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11 pages, 4116 KiB

Open AccessArticle

The Recent Advances in Raman Microscopy and Imaging Techniques for Biosensors

by Alexander Rzhevskii

Biosensors 2019, 9(1), 25; https://doi.org/10.3390/bios9010025 - 12 Feb 2019

Cited by 23 | Viewed by 8334

Abstract

Raman microspectroscopy is now well established as one of the most powerful analytical techniques for a diverse range of applications in physical (material) and biological sciences. Consequently, the technique provides exceptional analytical opportunities to the science and technology of biosensing due to its [...] Read more.

Raman microspectroscopy is now well established as one of the most powerful analytical techniques for a diverse range of applications in physical (material) and biological sciences. Consequently, the technique provides exceptional analytical opportunities to the science and technology of biosensing due to its capability to analyze both parts of a biosensor system—biologically sensitive components, and a variety of materials and systems used in physicochemical transducers. Recent technological developments in Raman spectral imaging have brought additional possibilities in two- and three-dimensional (2D and 3D) characterization of the biosensor’s constituents and their changes on a submicrometer scale in a label-free, real-time nondestructive method of detection. In this report, the essential components and features of a modern confocal Raman microscope are reviewed using the instance of Thermo Scientific DXRxi Raman imaging microscope, and examples of the potential applications of Raman microscopy and imaging for constituents of biosensors are presented. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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15 pages, 3631 KiB

Open AccessArticle

Detection of Metabolic Changes Induced via Drug Treatments in Live Cancer Cells and Tissue Using Raman Imaging Microscopy

by Mioara Larion, Tyrone Dowdy, Victor Ruiz-Rodado, Matthew W. Meyer, Hua Song, Wei Zhang, Dionne Davis, Mark R. Gilbert and Adrian Lita

Biosensors 2019, 9(1), 5; https://doi.org/10.3390/bios9010005 - 28 Dec 2018

Cited by 21 | Viewed by 7766

Abstract

Isocitrate dehydrogenase 1 (IDH1) mutations in gliomas, fibrosarcoma, and other cancers leads to a novel metabolite, D-2-hydroxyglutarate, which is proposed to cause tumorigenesis. The production of this metabolite also causes vulnerabilities in cellular metabolism, such as lowering NADPH levels. To exploit this vulnerability, [...] Read more.

Isocitrate dehydrogenase 1 (IDH1) mutations in gliomas, fibrosarcoma, and other cancers leads to a novel metabolite, D-2-hydroxyglutarate, which is proposed to cause tumorigenesis. The production of this metabolite also causes vulnerabilities in cellular metabolism, such as lowering NADPH levels. To exploit this vulnerability, we treated glioma and fibrosarcoma cells that harbor an IDH1 mutation with an inhibitor of nicotinamide adenine dinucleotide (NAD⁺) salvage pathway, FK866, and observed decreased viability in these cells. To understand the mechanism of action by which the inhibitor FK866 works, we used Raman imaging microscopy and identified that proteins and lipids are decreased upon treatment with the drug. Raman imaging showed a different distribution of lipids throughout the cell in the presence of the drug compared with the untreated cells. We employed nuclear magnetic resonance NMR spectroscopy and mass spectrometry to identify the classes of lipids altered. Our combined analyses point to a decrease in cell division due to loss of lipid content that contributes to membrane formation in the in vitro setting. However, the FK866 drug did not have the same potency in vivo. The use of Raman imaging microscopy indicated an opposite trend of lipid distribution in the tissue collected from treated versus untreated mice when compared with the cells. These results demonstrate the role of Raman imaging microscopy to identify and quantify metabolic changes in cancer cells and tissue. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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13 pages, 2194 KiB

Open AccessArticle

Simultaneous Detection of Dopamine and Serotonin—A Comparative Experimental and Theoretical Study of Neurotransmitter Interactions

by Felicia S. Manciu, Marian Manciu, John D. Ciubuc, Emma M. Sundin, Katia Ochoa, Michael Eastman, William G. Durrer, Jose Guerrero, Brayant Lopez, Mahendra Subedi and Kevin E. Bennet

Biosensors 2019, 9(1), 3; https://doi.org/10.3390/bios9010003 - 26 Dec 2018

Cited by 15 | Viewed by 8908

Abstract

With the goal of accurately detecting and quantifying the amounts of dopamine (DA) and serotonin (5-HT) in mixtures of these neurotransmitters without using any labelling, we present a detailed, comparative computational and Raman experimental study. Although discrimination between these two analytes is achievable [...] Read more.

With the goal of accurately detecting and quantifying the amounts of dopamine (DA) and serotonin (5-HT) in mixtures of these neurotransmitters without using any labelling, we present a detailed, comparative computational and Raman experimental study. Although discrimination between these two analytes is achievable in such mixtures for concentrations in the millimolar range, their accurate quantification remains unattainable. As shown for the first time in this work, the formation of a new composite resulting from their interactions with each other is the main reason for this lack of quantification. While this new hydrogen-bonded complex further complicates potential analyte discrimination and quantification at concentrations characteristic of physiological levels (i.e., nanomolar concentrations), it can also open new avenues for its use in drug delivery and pharmaceutical research. This remark is based not only on chemical interactions analyzed here from both theoretical and experimental approaches, but also on biological relationship, with consideration of both functional and neural proximity perspectives. Thus, this research constitutes an important contribution toward better understanding of neural processes, as well as toward possible future development of label-free biosensors. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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16 pages, 2236 KiB

Open AccessArticle

Label-Free SERS Discrimination and In Situ Analysis of Life Cycle in Escherichia coli and Staphylococcus epidermidis

by Niccolò Paccotti, Francesco Boschetto, Satoshi Horiguchi, Elia Marin, Alessandro Chiadò, Chiara Novara, Francesco Geobaldo, Fabrizio Giorgis and Giuseppe Pezzotti

Biosensors 2018, 8(4), 131; https://doi.org/10.3390/bios8040131 - 15 Dec 2018

Cited by 19 | Viewed by 6565

Abstract

Surface enhanced Raman spectroscopy (SERS) has been proven suitable for identifying and characterizing different bacterial species, and to fully understand the chemically driven metabolic variations that occur during their evolution. In this study, SERS was exploited to identify the cellular composition of Gram-positive [...] Read more.

Surface enhanced Raman spectroscopy (SERS) has been proven suitable for identifying and characterizing different bacterial species, and to fully understand the chemically driven metabolic variations that occur during their evolution. In this study, SERS was exploited to identify the cellular composition of Gram-positive and Gram-negative bacteria by using mesoporous silicon-based substrates decorated with silver nanoparticles. The main differences between the investigated bacterial strains reside in the structure of the cell walls and plasmatic membranes, as well as their biofilm matrix, as clearly noticed in the corresponding SERS spectrum. A complete characterization of the spectra was provided in order to understand the contribution of each vibrational signal collected from the bacterial culture at different times, allowing the analysis of the bacterial populations after 12, 24, and 48 h. The results show clear features in terms of vibrational bands in line with the bacterial growth curve, including an increasing intensity of the signals during the first 24 h and their subsequent decrease in the late stationary phase after 48 h of culture. The evolution of the bacterial culture was also confirmed by fluorescence microscope images. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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8 pages, 1207 KiB

Open AccessArticle

Biomolecular Component Analysis of Phospholipids Composition in Live HeLa Cells

by Svitlana M. Levchenko and Junle Qu

Biosensors 2018, 8(4), 123; https://doi.org/10.3390/bios8040123 - 5 Dec 2018

Cited by 6 | Viewed by 5418

Abstract

The alteration of the phospholipid composition within the cell, in particular the ratio between saturated and unsaturated fatty acids, can serve as an important biomarker to prognosis of the disease progression (e.g., fatty-liver disease, prostate cancer, or neurodegenerative disorders). Major techniques for lipid [...] Read more.

The alteration of the phospholipid composition within the cell, in particular the ratio between saturated and unsaturated fatty acids, can serve as an important biomarker to prognosis of the disease progression (e.g., fatty-liver disease, prostate cancer, or neurodegenerative disorders). Major techniques for lipid analysis in biological samples require a lipid extraction procedure that is not compatible with live cell studies. To address this challenge, we apply microRaman-Biomolecular Component Analysis (BCA) for comparative analysis of phospholipid composition and sensing the saturation degree of fatty acid lipid chain in live HeLa cells and lipids extracted from HeLa cells. After processing raw Raman data, acquired in lipid droplets (LDs) free cytoplasmic area, LDs and extracted lipids with BCA, the lipid component was isolated. Despite the similarity in general profiles of processed Raman spectra acquired in live cells and extracted lipids, some clear differences that reflect diversity in their phospholipids composition were revealed. Furthermore, using the direct relation between the number of double bonds in the fatty acid chain and the intensity ratio of the corresponding Raman bands, the saturation degree of fatty acids was estimated. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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11 pages, 1849 KiB

Open AccessFeature PaperArticle

BCAbox Algorithm Expands Capabilities of Raman Microscope for Single Organelles Assessment

by Andrey N. Kuzmin, Artem Pliss, Alex Rzhevskii, Adrian Lita and Mioara Larion

Biosensors 2018, 8(4), 106; https://doi.org/10.3390/bios8040106 - 10 Nov 2018

Cited by 18 | Viewed by 6686

Abstract

Raman microspectroscopy is a rapidly developing technique, which has an unparalleled potential for in situ proteomics, lipidomics, and metabolomics, due to its remarkable capability to analyze the molecular composition of live cells and single cellular organelles. However, the scope of Raman spectroscopy for [...] Read more.

Raman microspectroscopy is a rapidly developing technique, which has an unparalleled potential for in situ proteomics, lipidomics, and metabolomics, due to its remarkable capability to analyze the molecular composition of live cells and single cellular organelles. However, the scope of Raman spectroscopy for bio-applications is limited by a lack of software tools for express-analysis of biomolecular composition based on Raman spectra. In this study, we have developed the first software toolbox for immediate analysis of intracellular Raman spectra using a powerful biomolecular component analysis (BCA) algorithm. Our software could be easily integrated with commercial Raman spectroscopy instrumentation, and serve for precise analysis of molecular content in major cellular organelles, including nucleoli, endoplasmic reticulum, Golgi apparatus, and mitochondria of either live or fixed cells. The proposed software may be applied in broad directions of cell science, and serve for further advancement and standardization of Raman spectroscopy. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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16 pages, 14220 KiB

Open AccessArticle

Functionalized Gold Nanoparticles as Biosensors for Monitoring Cellular Uptake and Localization in Normal and Tumor Prostatic Cells

by Marianna Pannico, Anna Calarco, Gianfranco Peluso and Pellegrino Musto

Biosensors 2018, 8(4), 87; https://doi.org/10.3390/bios8040087 - 4 Oct 2018

Cited by 18 | Viewed by 4592

Abstract

In the present contribution the fabrication and characterization of functionalized gold nanospheres of uniform shape and controlled size is reported. These nano-objects are intended to be used as Surface Enhanced Raman Spectroscopy (SERS) sensors for in-vitro cellular uptake and localization. Thiophenol was used [...] Read more.

In the present contribution the fabrication and characterization of functionalized gold nanospheres of uniform shape and controlled size is reported. These nano-objects are intended to be used as Surface Enhanced Raman Spectroscopy (SERS) sensors for in-vitro cellular uptake and localization. Thiophenol was used as molecular reporter and was bound to the Au surface by a chemisorption process in aqueous solution. The obtained colloidal solution was highly stable and no aggregation of the single nanospheres into larger clusters was observed. The nanoparticles were incubated in human prostatic cells with the aim of developing a robust, SERS-based method to differentiate normal and tumor cell lines. SERS imaging experiments showed that tumor cells uptake considerably larger amounts of nanoparticles in comparison to normal cells (up to 950% more); significant differences were also observed in the uptake kinetics. This largely different behaviour might be exploited in diagnostic and therapeutic applications. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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10 pages, 4345 KiB

Open AccessArticle

Micro-Raman Spectroscopy for Monitoring of Deposition Quality of High-k Stack Protective Layer onto Nanowire FET Chips for Highly Sensitive miRNA Detection

by Kristina A. Malsagova, Tatyana O. Pleshakova, Andrey F. Kozlov, Ivan D. Shumov, Mikhail A. Ilnitskii, Andrew V. Miakonkikh, Vladimir P. Popov, Konstantin V. Rudenko, Alexander V. Glukhov, Igor N. Kupriyanov, Nina D. Ivanova, Alexander E. Rogozhin, Alexander I. Archakov and Yuri D. Ivanov

Biosensors 2018, 8(3), 72; https://doi.org/10.3390/bios8030072 - 27 Jul 2018

Cited by 15 | Viewed by 5377

Abstract

Application of micro-Raman spectroscopy for the monitoring of quality of high-k (h-k) dielectric protective layer deposition onto the surface of a nanowire (NW) chip has been demonstrated. A NW chip based on silicon-on-insulator (SOI) structures, protected with a layer of high-k dielectric ((h-k)-SOI-NW [...] Read more.

Application of micro-Raman spectroscopy for the monitoring of quality of high-k (h-k) dielectric protective layer deposition onto the surface of a nanowire (NW) chip has been demonstrated. A NW chip based on silicon-on-insulator (SOI) structures, protected with a layer of high-k dielectric ((h-k)-SOI-NW chip), has been employed for highly sensitive detection of microRNA (miRNA) associated with oncological diseases. The protective dielectric included a 2-nm-thick Al₂O₃ surface layer and a 8-nm-thick HfO₂ layer, deposited onto a silicon SOI-NW chip. Such a chip had increased time stability upon operation in solution, as compared with an unprotected SOI-NW chip with native oxide. The (h-k)-SOI-NW biosensor has been employed for the detection of DNA oligonucleotide (oDNA), which is a synthetic analogue of miRNA-21 associated with oncological diseases. To provide biospecificity of the detection, the surface of (h-k)-SOI-NW chip was modified with oligonucleotide probe molecules (oDVA probes) complementary to the sequence of the target biomolecule. Concentration sensitivity of the (h-k)-SOI-NW biosensor at the level of DL~10⁻¹⁶ M has been demonstrated. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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Review

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19 pages, 4242 KiB

Open AccessReview

Raman Spectroscopy and Microscopy Applications in Cardiovascular Diseases: From Molecules to Organs

by Ardalan Chaichi, Alisha Prasad and Manas Ranjan Gartia

Biosensors 2018, 8(4), 107; https://doi.org/10.3390/bios8040107 - 12 Nov 2018

Cited by 44 | Viewed by 9993

Abstract

Noninvasive and label-free vibrational spectroscopy and microscopy methods have shown great potential for clinical diagnosis applications. Raman spectroscopy is based on inelastic light scattering due to rotational and vibrational modes of molecular bonds. It has been shown that Raman spectra provide chemical signatures [...] Read more.

Noninvasive and label-free vibrational spectroscopy and microscopy methods have shown great potential for clinical diagnosis applications. Raman spectroscopy is based on inelastic light scattering due to rotational and vibrational modes of molecular bonds. It has been shown that Raman spectra provide chemical signatures of changes in biological tissues in different diseases, and this technique can be employed in label-free monitoring and clinical diagnosis of several diseases, including cardiovascular studies. However, there are very few literature reviews available to summarize the state of art and future applications of Raman spectroscopy in cardiovascular diseases, particularly cardiac hypertrophy. In addition to conventional clinical approaches such as electrocardiography (ECG), echocardiogram (cardiac ultrasound), positron emission tomography (PET), cardiac computed tomography (CT), and single photon emission computed tomography (SPECT), applications of vibrational spectroscopy and microscopy will provide invaluable information useful for the prevention, diagnosis, and treatment of cardiovascular diseases. Various in vivo and ex vivo investigations can potentially be performed using Raman imaging to study and distinguish pathological and physiological cardiac hypertrophies and understand the mechanisms of other cardiac diseases. Here, we have reviewed the recent literature on Raman spectroscopy to study cardiovascular diseases covering investigations on the molecular, cellular, tissue, and organ level. Full article

(This article belongs to the Special Issue Applications of Raman Techniques in Biosensing)

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