RESUMO
Raman spectroscopy is an important tool in understanding chemical components of various materials. However, the excessive weight and energy consumption of a conventional CCD-based Raman spectrometer forbids its applications under extreme conditions, including unmanned aircraft vehicles (UAVs) and Mars/Moon rovers. In this article, we present a highly sensitive, shot-noise-limited, and ruggedized Raman signal acquisition using a time-correlated photon-counting system. Compared with conventional Raman spectrometers, over 95% weight, 65% energy consumption, and 70% cost could be removed through this design. This technique allows space- and UAV-based Raman spectrometers to robustly perform hyperspectral Raman acquisitions without excessive energy consumption.
Assuntos
Fótons , Análise Espectral Raman/instrumentação , Análise Espectral Raman/métodos , Aeronaves , Algoritmos , Desenho de Equipamento , Exobiologia/instrumentação , Exobiologia/métodos , Marte , Lua , Tecnologia de Sensoriamento Remoto/instrumentação , Reprodutibilidade dos Testes , AstronaveRESUMO
We present a unique confocal microscope capable of measuring the Raman and Brillouin spectra simultaneously from a single spatial location. Raman and Brillouin scattering offer complementary information about a material's chemical and mechanical structure, respectively, and concurrent monitoring of both of these spectra would set a new standard for material characterization. We achieve this by applying recent innovations in Brillouin spectroscopy that reduce the necessary acquisition times to durations comparable to conventional Raman spectroscopy while attaining a high level of spectral accuracy. To demonstrate the potential of the system, we map the Raman and Brillouin spectra of a molded poly(ethylene glycol) diacrylate (PEGDA) hydrogel sample in cyclohexane to create two-dimensional images with high contrast at microscale resolutions. This powerful tool has the potential for very diverse analytical applications in basic science, industry, and medicine.
Assuntos
Técnicas de Química Analítica/métodos , Cicloexanos/química , Microscopia Confocal , Análise Espectral Raman , Fenômenos MecânicosRESUMO
Fast and sensitive Raman spectroscopy measurements are imperative for a large number of applications in biomedical imaging, remote sensing and material characterization. In this report, by introducing an electronically-tunable acousto-optical filter as a wavelength selector, we demonstrated a novel instrumentation to the broadband coherent Raman spectroscopy. System's tunability allows assessing Raman transitions ranging from <400 cm(-1) to 4500 cm(-1). We validated the use of the new instrumentation by collecting coherent anti-Stokes spectra and stimulated Raman spectra of various samples.
RESUMO
A novel concept of Brillouin imaging and sensing via time-resolved optical (BISTRO) measurements is introduced for flow cytometry applications. The system affords robust, maintenance-free and high-speed elasticity-sensitive measurements.
Assuntos
Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Imagem Óptica/instrumentação , Imagem Óptica/métodos , Espectrofotometria/métodos , Acústica , Anemia Falciforme/patologia , Animais , Elasticidade , Desenho de Equipamento , Eritrócitos/citologia , Humanos , Luz , Neoplasias/patologia , Fenótipo , Fônons , Refratometria , Espalhamento de Radiação , Processamento de Sinais Assistido por Computador , Espectrofotometria/instrumentaçãoRESUMO
Brillouin spectroscopy is an emerging tool for microscopic optical imaging as it allows for non-contact, non-invasive, direct assessment of the elastic properties of materials. However, strong elastic scattering and stray light from various sources often contaminate the Brillouin spectrum. A molecular absorption cell was introduced into the virtually imaged phased array (VIPA) based Brillouin spectroscopy setup to absorb the Rayleigh component, which resulted in a substantial improvement of the Brillouin spectrum quality.
Assuntos
Técnicas de Imagem por Elasticidade/métodos , Análise Espectral/métodos , Diagnóstico por Imagem/instrumentação , Diagnóstico por Imagem/métodos , Diagnóstico por Imagem/estatística & dados numéricos , Técnicas de Imagem por Elasticidade/instrumentação , Técnicas de Imagem por Elasticidade/estatística & dados numéricos , Humanos , Lasers de Estado Sólido , Luz , Dispositivos Ópticos , Fenômenos Ópticos , Espalhamento de Radiação , Análise Espectral/instrumentação , Análise Espectral/estatística & dados numéricos , Interface Usuário-ComputadorRESUMO
Stimulated Raman scattering (SRS) microscopy is a powerful tool for chemically-sensitive non-invasive optical imaging. However, ultrafast laser sources, which are currently employed, are still expensive and require substantial maintenance to provide temporal overlap and spectral tuning. SRS imaging, which utilizes continuous-wave laser sources, has a major advantage, as it eliminates the cell damage due to exposure to the high-intensity light radiation, while substantially reducing the cost and complexity of the set-up. As a proof-of-principle, we demonstrate microscopic imaging of dimethyl sulfoxide using two independent, commonly used lasers, a diode-pumped, intracavity doubled 532-nm laser and a He-Ne laser operating at 632.8-nm.
RESUMO
Antihistamines, which are commonly used to treat allergic reactions, are known for their side effects, which contribute to weight gain. It is hypothesized that simultaneous Brillouin elastography and Raman spectroscopy can be used to detect changes in adipose tissue associated with a prolonged intake of desloratadine, a commonly used second generation antihistamine. White and brown adipose tissue samples were excised from adult rats following 16 weeks of daily administration of desloratadine. It was found that the prolonged intake of desloratadine leads to an increase in Brillouin shift in both adipose tissue types. Raman spectra indicate that antihistamine use reduces protein-to-lipid ratio in brown adipose tissue but not white adipose tissue, indicating the effect on adipose tissue is location-dependent.
Assuntos
Antagonistas não Sedativos dos Receptores H1 da Histamina , Tecido Adiposo Marrom , Tecido Adiposo Branco , Animais , Antagonistas não Sedativos dos Receptores H1 da Histamina/farmacologia , Loratadina/análogos & derivados , Loratadina/farmacologia , RatosRESUMO
Brillouin spectroscopy is an emerging tool in biomedical imaging and sensing. It is capable of assessing the high-frequency viscoelastic longitudinal modulus with microscopic spatial resolution. Nonlinear Brillouin spectroscopy based on impulsive stimulated Brillouin scattering offers a number of significant advantages over conventional spontaneous and stimulated Brillouin scattering. In this report, we evaluate the accuracy of Brillouin shift measurements in spontaneous and nonlinear Brillouin microscopy by calculating the Allan variance for both CW excited spontaneous Brillouin measurements and nonlinear Brillouin scattering measurements made with both nanosecond and picosecond pulse excitation. We find that impulsive stimulated Brillouin spectroscopy is superior to spontaneous Brillouin spectroscopy in terms of the accuracy of such measurements and demonstrate its application for assessing tiny changes in Brillouin frequency shifts associated with low concentrations of biologically relevant solutions.
RESUMO
The main objective of the present study is to evaluate the use of Brillouin microspectroscopy for differentiation of melanoma and normal tissues based on elasticity measurements. Previous studies of malignant melanoma show that the lesion is stiffer than the surrounding healthy tissue. We hypothesize that elasticity-specific Brillouin spectroscopy can be used to distinguish between healthy and cancerous regions of an excised melanoma from a Sinclair miniature swine. Brillouin measurements of non-regressing and regressing melanomas and the surrounding healthy tissues were performed. Based on the Brillouin measurements, the melanomas and healthy tissues can be successfully differentiated. The stiffness of both tumors is found to be significantly greater than the healthy tissues. Notably, we found that the elasticity of regressing melanoma is closer to that of the normal tissue. The results indicate that Brillouin spectroscopy can be utilized as a tool for elasticity-based differentiation between malignant melanoma and surrounding healthy tissue, with potential use for melanoma boundary identification, monitoring tumor progression, or response to treatment.
RESUMO
Our current understanding of the mechanical properties of nanostructured biomaterials is rather limited to invasive/destructive and low-throughput techniques such as atomic force microscopy, optical tweezers, and shear rheology. In this report, we demonstrate the capabilities of recently developed dual Brillouin/Raman spectroscopy to interrogate the mechanical and chemical properties of nanostructured hydrogel networks. The results obtained from Brillouin spectroscopy show an excellent correlation with the conventional uniaxial and shear mechanical testing. Moreover, it is confirmed that, unlike the macroscopic conventional mechanical measurement techniques, Brillouin spectroscopy can provide the elasticity characteristic of biomaterials at a mesoscale length, which is remarkably important for understanding complex cell-biomaterial interactions. The proposed technique experimentally demonstrated the capability of studying biomaterials in their natural environment and may facilitate future fabrication and inspection of biomaterials for various biomedical and biotechnological applications.
Assuntos
Materiais Biocompatíveis/química , Hidrogéis/química , Nanopartículas/química , Nanopartículas/ultraestrutura , Engenharia TecidualRESUMO
Obesity is becoming a leading cause of health problems world-wide. Obesity and overweight are associated with the structural and chemical changes in tissues; however, few methods exist that allow for concurrent measurement of these changes. Using Brillouin and Raman microspectroscopy, both the mechanical and chemical differences can be assessed simultaneously. We hypothesized that Brillouin spectroscopy can measure the adipose tissues' stiffness, which increases in obesity. Samples of brown and white adipose tissues obtained from control and diet-induced obese adult rats were analyzed. The results show that both adipose tissues of the obese group exhibit a greater high-frequency longitudinal elastic modulus than the control samples, and that the brown fat is generally stiffer than white adipose. The Raman spectra indicate that the lipids' accumulation in adipose tissue outpaces the fibrosis, and that the high-fat diet has a greater effect on the brown adipose than the white fat. Overall, the powerful combination of Brillouin and Raman microspectroscopies successfully assessed both the mechanical properties and chemical composition of adipose tissue simultaneously for the first time. The results indicate that the adipose tissue experiences an obesity-induced increase in stiffness and lipid content, with the brown adipose tissue undergoing a more pronounced change compared to white adipose.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Dieta Hiperlipídica/efeitos adversos , Fenômenos Mecânicos , Obesidade/induzido quimicamente , Obesidade/patologia , Fenômenos Ópticos , Tecido Adiposo/patologia , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Marrom/patologia , Tecido Adiposo Branco/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Tecido Adiposo Branco/patologia , Animais , Fenômenos Biomecânicos/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Brillouin spectroscopy is an emerging tool for microscopic optical imaging as it allows noninvasive assessment of viscoelastic properties of materials. The use of atomic-molecular absorption cells as ultra-narrow notch filters allows acquisition of Brillouin spectra from turbid samples despite their strong elastic scattering. However, such systems alter the shapes of the Brillouin lines, making the precise determination of the Brillouin shift difficult. In this report, we propose a simple method for analyzing the Brillouin spectrum using a customized least-square fitting algorithm. The absorption spectrum induced by the atomic-molecular cell was taken into consideration. The capability of the method is confirmed by processing experimental spectroscopic data from the pure water at different temperatures. The accuracy of the measurements of ±1 MHz spectral line shift is experimentally demonstrated.
RESUMO
Fast and sensitive Raman spectroscopy measurements are imperative for a large number of applications in biomedical imaging, remote sensing and material characterization. Stimulated Raman spectroscopy offers a substantial improvement in the signal-to-noise ratio but is often limited to a discrete number of wavelengths. In this report, by introducing an electronically-tunable acousto-optical filter as a wavelength selector, a novel approach to a broadband stimulated Raman spectroscopy is demonstrated. The corresponding Raman shift covers the spectral range from 600 cm(-1) to 4500 cm(-1), sufficient for probing most vibrational Raman transitions. We validated the use of the new instrumentation to both coherent anti-Stokes scattering (CARS) and stimulated Raman scattering (SRS) spectroscopies.
RESUMO
Brillouin microspectroscopy is a powerful technique for noninvasive optical imaging. In particular, Brillouin microspectroscopy uniquely allows assessing a sample's mechanical properties with microscopic spatial resolution. Recent advances in background-free Brillouin microspectroscopy make it possible to image scattering samples without substantial degradation of the data quality. However, measurements at the cellular- and subcellular-level have never been performed to date due to the limited signal strength. In this report, by adopting our recently optimized VIPA-based Brillouin spectrometer, we probed the microscopic viscoelasticity of individual red blood cells. These measurements were supplemented by chemically specific measurements using Raman microspectroscopy.
Assuntos
Espaço Intracelular , Fenômenos Mecânicos , Análise Espectral Raman/métodos , Animais , Fenômenos Biomecânicos , Bovinos , Eritrócitos/citologia , MicroscopiaRESUMO
Bacterial meningitis is a disease of pronounced clinical significance, especially in the developing world. Immediate treatment with antibiotics is essential, and no single test can provide a conclusive diagnosis. It is well established that elevated total protein in cerebrospinal fluid (CSF) is associated with bacterial meningitis. Brillouin spectroscopy is a widely used optical technique for noninvasive determination of the elastic moduli of materials. We found that elevated protein levels in CSF alter the fluid elasticity sufficiently to be measurable by Brillouin spectroscopy, with model healthy and diseased fluids distinguishable to marked significance (P = 0.014), which increases with sample concentration by dialysis. Typical raw output of a 2-stage VIPA Brillouin spectrometer: inelastically scattered Brillouin peaks (arrows) and elastically scattered incident radiation (center cross).
Assuntos
Proteínas do Líquido Cefalorraquidiano/análise , Meningites Bacterianas/líquido cefalorraquidiano , Análise Espectral/métodos , Proteínas do Líquido Cefalorraquidiano/química , Humanos , Espectrometria de Fluorescência , Análise Espectral Raman , Fatores de TempoRESUMO
Two-dimensional stimulated Brillouin scattering microscopy is demonstrated for the first time using low power continuous-wave lasers tunable around 780 nm. Spontaneous Brillouin spectroscopy has much potential for probing viscoelastic properties remotely and non-invasively on a microscopic scale. Nonlinear Brillouin scattering spectroscopy and microscopy may provide a way to tremendously accelerate the data aquisition and improve spatial resolution. This general imaging setup can be easily adapted for specific applications in biology and material science. The low power and optical wavelengths in the water transparency window used in this setup provide a powerful bioimaging technique for probing the mechanical properties of hard and soft tissue.