Spectral fiber photometry derives hemoglobin concentration changes for accurate measurement of fluorescent sensor activity.
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ABSTRACT:
SUBMITTER: Zhang WT
PROVIDER: S-EPMC9308135 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:This study examines the use of a conductive carbon fiber to construct a flexible biosensing platform for monitoring biomarkers in sweat. Cortisol was chosen as a model analyte. Functionalization of the conductive carbon yarn (CCY) with ellipsoidal Fe2O3 has been performed to immobilize the antibodies specific to cortisol. 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-Hydroxysuccinimide (NHS) chemistry has been used to immobilize the antibodies onto the Fe2O3 modified CCY. Crystallinity, structure, morphology, flexibility, surface area, and elemental analysis were studied using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Raman spectroscopy, Field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FE-SEM/EDS) and Brunauer-Emmett-Teller (BET) analysis. Mechanical properties of the fiber such as tensile strength, young's modulus have also been investigated. Under optimal parameters, the fabric sensor exhibited a good linearity (r2?=?0.998) for wide a linear range from 1 fg to 1??g with a detection limit of 0.005 fg/mL for the sensitive detection of cortisol. Repeatability, reliability, reproducibility, and anti-interference properties of the current sensor have been investigated. Detection of cortisol levels in human sweat samples has also been investigated and the results were validated with commercial chemiluminescence immunoassay (CLIA) method.
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Project description:Fiber photometry is a key technique for characterizing brain-behavior relationships in vivo. Initially, it was primarily used to report calcium dynamics as a proxy for neural activity via genetically encoded indicators. This generated new insights into brain functions including movement, memory, and motivation at the level of defined circuits and cell types. Recently, the opportunity for discovery with fiber photometry has exploded with the development of an extensive range of fluorescent sensors for biomolecules including neuromodulators and peptides that were previously inaccessible in vivo. This critical advance, combined with the new availability of affordable "plug-and-play" recording systems, has made monitoring molecules with high spatiotemporal precision during behavior highly accessible. However, while opening exciting new avenues for research, the rapid expansion in fiber photometry applications has occurred without coordination or consensus on best practices. Here, we provide a comprehensive guide to help end-users execute, analyze, and suitably interpret fiber photometry studies.
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Project description:Ammonium (NH??) is the most common N-source for yeast fermentations, and N-limitation is frequently applied to reduce growth and increase product yields. While there is significant molecular knowledge on NH?? transport and assimilation, there have been few attempts to measure the in vivo concentration of this metabolite. In this article, we present a sensitive and accurate analytical method to quantify the in vivo intracellular ammonium concentration in Saccharomyces cerevisiae based on standard rapid sampling and metabolomics techniques. The method validation experiments required the development of a proper sample processing protocol to minimize ammonium production/consumption during biomass extraction by assessing the impact of amino acid degradation-an element that is often overlooked. The resulting cold chloroform metabolite extraction method, together with quantification using ultra high performance liquid chromatography-isotope dilution mass spectrometry (UHPLC-IDMS), was not only more sensitive than most of the existing methods but also more accurate than methods that use electrodes, enzymatic reactions, or boiling water or boiling ethanol biomass extraction because it minimized ammonium consumption/production during sampling processing and interference from other metabolites in the quantification of intracellular ammonium. Finally, our validation experiments showed that other metabolites such as pyruvate or 2-oxoglutarate (?KG) need to be extracted with cold chloroform to avoid measurements being biased by the degradation of other metabolites (e.g., amino acids).
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Project description:BackgroundHand hygiene is a crucial and cost-effective method to prevent health care-associated infections, and in 2009, the World Health Organization (WHO) issued guidelines to encourage and standardize hand hygiene procedures. However, a common challenge in health care settings is low adherence, leading to low handwashing quality. Recent advances in machine learning and wearable sensing have made it possible to accurately measure handwashing quality for the purposes of training, feedback, or accreditation.ObjectiveWe measured the accuracy of a sensor armband (Myo armband) in detecting the steps and duration of the WHO procedures for handwashing and handrubbing.MethodsWe recruited 20 participants (10 females; mean age 26.5 years, SD 3.3). In a semistructured environment, we collected armband data (acceleration, gyroscope, orientation, and surface electromyography data) and video data from each participant during 15 handrub and 15 handwash sessions. We evaluated the detection accuracy for different armband placements, sensor configurations, user-dependent vs user-independent models, and the use of bootstrapping.ResultsUsing a single armband, the accuracy was 96% (SD 0.01) for the user-dependent model and 82% (SD 0.08) for the user-independent model. This increased when using two armbands to 97% (SD 0.01) and 91% (SD 0.04), respectively. Performance increased when the armband was placed on the forearm (user dependent: 97%, SD 0.01; and user independent: 91%, SD 0.04) and decreased when placed on the arm (user dependent: 96%, SD 0.01; and user independent: 80%, SD 0.06). In terms of bootstrapping, user-dependent models can achieve more than 80% accuracy after six training sessions and 90% with 16 sessions. Finally, we found that the combination of accelerometer and gyroscope minimizes power consumption and cost while maximizing performance.ConclusionsA sensor armband can be used to measure hand hygiene quality relatively accurately, in terms of both handwashing and handrubbing. The performance is acceptable using a single armband worn in the upper arm but can substantially improve by placing the armband on the forearm or by using two armbands.
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Project description:In vivo electrophysiology is the gold standard technique used to investigate sub-second neural dynamics in freely behaving animals. However, monitoring cell-type-specific population activity is not a trivial task. Over the last decade, fiber photometry based on genetically encoded calcium indicators (GECIs) has been widely adopted as a versatile tool to monitor cell-type-specific population activity in vivo. However, this approach suffers from low temporal resolution. Here, we combine these two approaches to monitor both sub-second field potentials and cell-type-specific population activity in freely behaving mice. By developing an economical custom-made system and constructing a hybrid implant of an electrode and a fiber optic cannula, we simultaneously monitor artifact-free mesopontine field potentials and calcium transients in cholinergic neurons across the sleep-wake cycle. We find that mesopontine cholinergic activity co-occurs with sub-second pontine waves, called P-waves, during rapid eye movement sleep. Given the simplicity of our approach, simultaneous electrophysiological recording and cell-type-specific imaging provides a novel and valuable tool for interrogating state-dependent neural circuit dynamics in vivo.
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Project description:We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N'-Dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations <50μM. Using nasal inhalation of the sensor and spectral unmixing to generate images, E149C/A213R GBP-IANBD fluorescence was detected in luminal regions of cryosections of the murine distal lung that was greater in db/db than wildtype mice. In conclusion, this sensor provides a useful tool for further development to measure luminal glucose concentration in models of lung/airway to explore how this may change in disease.
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Project description:We conducted a study to assess the pressure difference between the aqueous and vitreous humors in rabbit eyes using a direct intraocular pressure (IOP) measurement method. A micro-optic-fiber pressure sensor was utilized for this purpose. Preliminary experiments with enucleated porcine eyes confirmed the sensor's accuracy in measuring both aqueous and vitreous humor pressure. The main study involved six healthy albino rabbits, where the sensor measured the pressure in the anterior chamber (aIOP) and posterior vitreous-cavity (pIOP). These measurements were compared to aIOP values obtained through rebound tonometry. Additionally, pre- and postoperative pressure comparisons were made after performing a vitrectomy. Results revealed a significant disparity between aqueous and vitreous humor pressures. Prior to vitrectomy, pIOP was 22.8 mmHg, over twice as high as aIOP (11.0 mmHg), but decreased to a similar level following the procedure. Comparison between the sensor measurements and rebound tonometry showed agreement in aIOP values. In conclusion, our study demonstrates that vitreous humor pressure is consistently higher than aqueous humor pressure, reaching the upper limit of normal IOP. Furthermore, vitrectomy effectively reduces pIOP, aligning it with aIOP. These findings contribute valuable insights into intraocular pressure dynamics and have implications for clinical interventions targeting ocular pressure regulation.
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Project description:In this work, a linear birefringence measurement method is proposed for an optical fiber current sensor (OFCS). First, the optical configuration of the measurement system is presented. Then, the elimination method of the effect of the azimuth angles between the sensing fiber and the two polarizers is demonstrated. Moreover, the relationship of the linear birefringence, the Faraday rotation angle and the final output is determined. On these bases, the multi-valued problem on the linear birefringence is simulated and its solution is illustrated when the linear birefringence is unknown. Finally, the experiments are conducted to prove the feasibility of the proposed method. When the numbers of turns of the sensing fiber in the OFCS are about 15, 19, 23, 27, 31, 35, and 39, the measured linear birefringence obtained by the proposed method are about 1.3577, 1.8425, 2.0983, 2.5914, 2.7891, 3.2003 and 3.5198 rad. Two typical methods provide the references for the proposed method. The proposed method is proven to be suitable for the linear birefringence measurement in the full range without the limitation that the linear birefringence must be smaller than π/2.
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