Liquid Biological Samples Research Articles

Automatic characterization of capillary flow profile of liquid samples on μTADs based on capacitance measurement

Capillary flow profile of liquid samples in porous media is closely related to the important properties of liquid samples, including the viscosity and the surface energy. Therefore, capillary flow profile can be used as an index to differentiate liquid samples with different properties. Fast and automatic characterization of capillary flow profile of liquid samples is necessary. In this work, we develop a portable and economical capacitance acquisition system (CASY) to easily obtain the capillary flow profile of liquid samples on microfluidic thread-based analytical devices (μTADs) by measuring the capacitance during the capillary flow. At first, we validate the accuracy of this method by comparing with the traditional method by video analysis in obtaining the capillary flow profiles in μTADs of cotton threads or glass fiber threads. Then we use it to differentiate liquid samples with different viscosity (mixture of water and glycerol). In addition, capillary flow profile on μTADs with chemical valves (chitosan or sucrose) can also be obtained on this device. Lastly, we show the potential of this device in measurement of hematocrit (HCT) of whole blood samples. This device can be used to catalog liquid biological samples with different properties in point-of-care diagnostics in the near future.

Simultaneous Determination of Multi-Class Mushroom Toxins in Mushroom and Biological Liquid Samples Using LC-MS/MS

A comprehensive analytical method based on liquid chromatography–tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous detection of 12 mushroom toxins (ibotenic acid, muscimol, muscarine, β-amanitin, α-amanitin, desoxoviroidin, γ-amanitin, phallisacin, illudin S, phallacidin, phalloidin and illudin M) in mushrooms, serum, urine and simulated gastric fluid. The samples were extracted with water or acetonitrile solution, and the serum sample was further purified with PSA sorbent. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 column with gradient elution using methanol and water containing 1 mM ammonia fluoride as a mobile phase. Mass spectrometric acquisition was performed in electrospray positive ionization mode. Good linearities (R2 > 0.994) were obtained for 12 toxins over the range of 0.05~200 µg/L. Matrix-matched calibration curves were used for quantification. The method limits of quantification were 0.01~0.2 mg/kg for mushrooms and 0.15~2.0 µg/L for three biological liquid samples. The mean recoveries of 12 target toxins (spiked at three concentration levels) ranged from 73.0% to 110.3%, with relative standard deviations not exceeding 19.4%, which meets the requirements for the determination of trace compounds in a biological matrix. This method was applied to the analysis of mushroom samples from Yunnan Province. As a result, 11 toxins, not including illudin M, were detected with a concentration range of 0.61~2143 mg/kg.

A disposable optofluidic micro-transmission cell with tailorable length for Fourier-transform infrared spectroscopy of biological fluids.

Mid-infrared Fourier-transform infrared (FT-IR) spectroscopy of liquid biological samples is limited by the high absorption of water in this spectral range, which makes conventional transmission cuvettes unsuitable as their centimeter-scale length is already too big. The most common alternative relies on the use of attenuated total reflection (ATR) accessories, which provide a small interaction path length for light along the interface between the analyte and the expensive ATR crystals. In this work, we address this issue by proposing a disposable and low-cost micro-transmission cell. Its construction relies on a simple technique, which consists of dispersing plastic spherical microparticles in a liquid sample before dispensing it between two pieces of silicon assembled one onto the other and acting as windows for the cell. Consequently, the microparticles act as a spacer of very precise height in-between the two silicon windows. This technique allows easy construction of infrared absorption cells with near-optimum optical interaction path length just by selecting the most appropriate particle size. The concept is demonstrated by measuring the concentration of glucose in aqueous solutions using microspheres of diameter 20 μm then 40 μm and analyzing the corresponding glucose absorption peaks in the wavenumber range 950-1200 cm-1. The performance is compared to that of standard ATR spectroscopy of the same samples. This resulted in a root-mean-square error of cross-validation (RMSECV) of 58.8 mg dl-1 as obtained for transmission measurements by partial least squares (PLS) regression, which is comparable to the RMSECV of 53 mg dl-1 for single-reflection diamond ATR measurements.

Visualizing Molecular Dynamics by High-Speed Atomic Force Microscopy.

Dynamic processes and structural changes of biological molecules are essential to life. While conventional atomic force microscopy (AFM) is able to visualize molecules and supramolecular assemblies at sub-nanometer resolution, it cannot capture dynamics because of its low imaging rate. The introduction of high-speed atomic force microscopy (HS-AFM) solved this problem by providing a large increase in imaging velocity. Using HS-AFM, one is able to visualize dynamic molecular events with high spatiotemporal resolution under near-to physiological conditions. This approach opened new windows as finally dynamics of biomolecules at sub-nanometer resolution could be studied. Here we describe the working principles and an operation protocol for HS-AFM imaging and characterization of biological samples in liquid.

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats during EEG Recordings.

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

Detection of the minimum concentrations of α-lactose solution using high-power THz-ATR spectroscopy.

Terahertz (THz) technology has emerged as a promising tool for the qualitative and quantitative identification of markers containing major diseases, enabling early diagnosis and staged treatment of diseases. Nevertheless, the detection of water-containing biological samples is facing significant challenges due to limitations in high-power THz radiation sources and high-sensitivity detection devices. In this paper, we present a designed and constructed set of Terahertz-Attenuated Total Reflection (THz-ATR) spectrometer for high-sensitivity detection of liquid biological samples, which can dynamically maintain the signal-to-noise ratio (SNR) of THz detection signal of liquid biological samples at 40-60dB. Our high-power THz-ATR spectroscopy can identify and quantitatively detect α-lactose aqueous solution with a minimum concentration of 0.292mol/L. Moreover, we observed that the rate of change in the absorption peak position varied greatly between high and low concentration samples. Our high-power, high-sensitivity THz-ATR spectroscopy detection provides a rapid, accurate, and low-cost method for detecting disease markers such as blood and urine indicators. Additionally, this approach offers new perspectives for the refinement and in-depth detection of biomedical samples.

Detection and quantification of heavy metals in blood and milk of Amazon river dolphin (Inia geoffrensis) (Cetartiodactyla: Iniidae) using wavelength‐dispersive X‐ray fluorescence spectrometry

AbstractRiver dolphins of the genus Inia are among the most endangered cetacean species in the world. However, information on the elemental constitution of the blood and milk of these species is still quite scarce. This study aims to evaluate the wavelength‐dispersive X‐ray fluorescence (WD‐XRF) spectrometry to detect and quantify heavy metals in blood and milk samples of a free‐ranging female Amazon river dolphin (Inia geoffrensis) from Central Amazon, Brazil. The WD‐XRF method detected and quantified 14 elements (Na, Mg, Al, Si, P, S, Cl, K, Ca, Cr, Mn, Fe, Zn, and Cu) in the samples analyzed, in which 13 elements were in the blood and 14 were in the milk. The WD‐XRF method did not detect some heavy metals, including those already described for the Amazon river dolphin milk (e.g., mercury) detected by other analytical methods, such as atomic absorption spectrometry (AAS). However, it is possible that the samples analyzed may come from a non‐contaminated individual. Therefore, the joint use of different analytical methods is recommended for future analyses. In addition, both blood and milk samples presented high concentrations of some metals, and the health risks of botos are discussed. This result demonstrates that the WD‐XRF method is a viable analytical procedure for detecting certain elements and quantifying their concentrations in liquid biological samples of Amazonian cetaceans.

Microfluidic Liquid Cell with Silicon Nitride Super-Thin Membrane for Electron Microscopy of Samples in Liquid.

Microfluidic liquid cells have been developed to visualize nanoscaled biological samples in liquid using a scanning electron microscope (SEM) through an electron-transparent membrane (ETM). However, despite the combination of the high-resolution visualization of SEM and the high experimental capability of microfluidics, the image is unclear because of the scattering of the electron beam in the ETM. Thus, this study developed a microfluidic liquid cell with a super-thin ETM of thickness 10 nm. Because the super-thin ETM is excessively fragile, the bonding of a silicon-nitride-deposited substrate and a polydimethylsiloxane microchannel before silicon anisotropic etching was proposed prevented the super-thin ETM from damage and breakage due to etching. With this protection against etchant using the microchannel, the yield of the fabricated super-thin ETM increased from 0 to 87%. Further, the scattering of the electron beam was suppressed using a microfluidic liquid cell with a super-thin ETM, resulting in high-resolution visualization. In addition, T4 bacteriophages were visualized using a super-thin ETM in vacuum. Furthermore, the cyanobacterium Synechocystis sp. PCC6803 in liquid was visualized using a super-thin ETM, and sub-microscopic structures on the surface were observed.

Using infrared imaging for surface dosimetric measurements during microwaves exposures

Infrared (IR) thermography enabled by a small camera attached to a mobile phone is used in the present research for the determination of the heating dynamics of biological liquid samples exposed to 2.3 GHz continuous wave. The methodology is presented and analysed and some relevant examples are Figured out. Differences in electromagnetic field dissipation are nicely emphasized proving that microwaves dosimetry applied to surfaces, if enough resolution and frame frequency is enabled, may represent a valuable tool in observing heating dynamics of the exposed surfaces even at the microscale.

Approaches to Formaldehyde Measurement: From Liquid Biological Samples to Cells and Organisms.

Formaldehyde (FA) is the simplest aldehyde present both in the environment and in living organisms. FA is an extremely reactive compound capable of protein crosslinking and DNA damage. For a long time, FA was considered a “biochemical waste” and a by-product of normal cellular metabolism, but in recent decades the picture has changed. As a result, the need arose for novel instruments and approaches to monitor and measure not only environmental FA in water, cosmetics, and household products, but also in food, beverages and biological samples including cells and even organisms. Despite numerous protocols being developed for in vitro and in cellulo FA assessment, many of them have remained at the “proof-of-concept” stage. We analyze the suitability of different methods developed for non-biological objects, and present an overview of the recently developed approaches, including chemically-synthesized probes and genetically encoded FA-sensors for in cellulo and in vivo FA monitoring. We also discuss the prospects of classical methods such as chromatography and spectrophotometry, and how they have been adapted in response to the demand for precise, selective and highly sensitive evaluation of FA concentration fluctuations in biological samples. The main objectives of this review is to summarize data on the main approaches for FA content measurement in liquid biological samples, pointing out the advantages and disadvantages of each method; to report the progress in development of novel molecules suitable for application in living systems; and, finally, to discuss genetically encoded FA-sensors based on existing natural biological FA-responsive elements.

Relationship between delayed luminescence emission and mitochondrial status in Saccharomyces cerevisiae

Delayed luminescence (DL) is gradually used in various detection of biological systems as a rapid detection technique, however, its biological mechanism was still not clear. In this study, a new model of DL detection system for liquid biological samples is established to investigate the DL emission of Saccharomyces cerevisiae cells cultured in different glucose concentrations. We analyzed the relationship between the DL emission and cell growth, cell vitality, mitochondrial morphology, mitochondrial DNA (mtDNA) copy number, adenosine triphosphate (ATP), oxygen consumption rate (OCR), as well as mitochondria membrane potential (MMP) in S. cerevisiae cells cultured with 0.01, 0.05, 0.15, 3, 10 and 20 g/L glucose respectively. It was found that the DL emission had strong correlation with mitochondrial morphology, OCR, and MMP. The results suggested that DL is an indicator of mitochondria status under different glucose supply conditions, and may be an effective method to detect mitochondrial metabolism related disorders.

2D Nanomaterial-Based Hybrid Structured (Au-WSe2-PtSe2-BP) Surface Plasmon Resonance (SPR) Sensor With Improved Performance

As a promising optical method used in a variety of applications surface plasmon resonance (SPR) sensors are employed over a wide range of boundaries. This research proposes a highly sensitive SPR based sensor with a novel hybrid structure using transition metal dichalcogenides (e.g. WSe <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> , PtSe <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> ) along with black phosphorene (BP) through comprehensive numerical study. To analyze and evaluate the performances of the proposed sensor, the widely used transfer matrix method (TMM) was used. The performances of the sensor were measured in terms of reflectivity, sensitivity, detection accuracy (DA), and figure of merit (FOM). The sensor structure was optimized by changing different structural parameters of the hybrid architecture to obtain better performances. The results revealed that insertion of PtSe <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> with WSe <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> and BP over a gold layer of the conventional structure improved the performance of the sensor and the maximum sensitivity of the sensor was measured as 200 deg/RIU with a FOM of 17.70 RIU <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−1</sup> . As well, the light penetration through the optimized sensor is investigated using the finite element method (FEM) based software. With this kind of high sensing capabilities, it may be convinced that the proposed sensor can be applied in different fields of biosensing to detect liquid biological and biochemical samples or analytes.

Terahertz spectra of electrolyte solutions under applied electric and magnetic fields

Most biomolecules require an aqueous environment to fully exert their biological activity. However, the rotation mode, vibration mode, and energy associated with the hydrogen bonding network of water are in the terahertz band, resulting in strong absorption. Therefore, it is difficult to detect liquid biological samples using the terahertz technology. Here, a high-transmittance double-layer microfluidic chip was prepared using a cycloolefin copolymer material with high transmittance of terahertz waves. Combined with terahertz time-domain spectroscopy, the terahertz spectral characteristics of deionized water, NaCl, NaCO3, and CH3COONa solutions were studied. The changes in the terahertz transmission intensity of these electrolyte solutions under constant electric and magnetic fields were measured. The results show that the terahertz spectra of different sodium salt solutions with the same concentration of 0.9 mol/L are different. Furthermore, the terahertz absorption coefficients of the different electrolyte solutions gradually decrease with the increase of their residence time under the electric field, which is contrary to the results obtained under the external magnetic field. This study provides a new idea for the detection of sodium salt solution and lays a foundation for the development of THz technology.

Nitrocellulose redox permanganometry: A simple method for reductive capacity assessment

We propose a rapid, simple, and robust method for measurement of the reductive capacity of liquid and solid biological samples based on potassium permanganate reduction followed by trapping of manganese dioxide precipitate on a nitrocellulose membrane. Moreover, we discuss how nitrocellulose redox permanganometry (NRP) can be used for high-throughput analysis of biological samples and present HistoNRP, its modification used for detailed analysis of reductive capacity spatial distribution in tissue with preserved anatomical relations.•NRP is a rapid, cost-effective, and simple method for reductive capacity assessment•NRP is compatible with a high-throughput screening of solid and liquid biological samples•HistoNRP exploits passive diffusion slice print blotting for reductive capacity spatial analysis

Designing, fabrication and evaluation of a rapid, point-of-care and noninvasive system for the detection of lead (Pb2+).

Non-invasive collection of biological sample such as sweat, urine, saliva, hairs and, stool and onsite detection of anlaytes in those samples is an interesting and viable approach for rapid screening of various toxicants in body. Environmental exposure/presence of lead (82Pb) and its rapid detection provide one such opportunity. A chemical spot based colorimetric method and a transdermal patch device based on this spot test, is developed for rapid and qualitative assessment of inorganic lead (Pb2+) in non-coloured biological or environmental liquid samples. The transdermal patch system contains two important parts, a chemical spot prepared on a thin glass sheet and, an absorbent paper (11µm pore size). A one step colour development reaction is able to identify the presence or absence of Pb2+. In-vitro evaluation for sensitivity and cut-off value determination, within run and between run precision testing, specificity testing were done. In-vivo evaluation of the developed patch system was performed in occupationally lead-exposed subjects and in control volunteers. In-vivo field testing results were further validated with gold standard test for lead detection. Blood lead levels and patch lead levels were found to be positively correlated (r = 0.57, P < 0.0001). In addition, the sensitivity and specificity of device in identification of Pb2+ was found to be 75.93% (95% CI = 62.36%-86.51%) and 95.24% (95% CI = 76.18%-99.88%). The developed system appears as a reliable, non-invasive rapid test with minimum step involve for identification of Pb2+ in a given system.

Ultrasmall Volume Single-Droplet Viscometry: Monitoring Cornering Instabilities on Omniphobic Polydimethylsiloxane Brushes.

Viscosity is an essential fluid property that is important for industrial and laboratory applications. For biological, complex, and/or precious liquid samples, the available volume of fluid is limited, yet there are few existing techniques to measure the viscosity of small volumes of liquids. We report a facile method to measure the viscosity of liquids by monitoring the sliding of single-cornered droplets on surfaces coated with an omniphobic film that minimizes the contact-angle hysteresis. The developed measurement method was capable of accurately characterizing the viscosity of various liquids and showed statistically equivalent values when compared to the literature, for fluids with viscosities ranging from 0.35 to ∼800 mPa s (acetone to castor oil). Using the developed single-droplet viscometer, the minimum volume required to measure the viscosity of hexadecane, dodecane, toluene, and ethanol was <5 μL and was <1 μL for decane and isopropyl alcohol, respectively. Further, the viscosity of hexadecane measured from 22 to 70 °C matched literature values precisely. The single-droplet, small-volume viscometer also requires minimal cleaning due to the omniphobic surface, meaning the fluid may be reused for other purposes with no liquid loss occurring due to the viscosity measurement.

Scanning ion-conductance microscopy with a double-barreled nanopipette for topographic imaging of charged chromosomes.

Scanning ion conductance microscopy (SICM) is useful for imaging soft and fragile biological samples in liquids because it probes the samples' surface topography by detecting ion currents under non-contact and force-free conditions. SICM acquires the surface topographical height by detecting the ion current reduction that occurs when an electrolyte-filled glass nanopipette approaches the sample surface. However, most biological materials have electrically charged surfaces in liquid environments, which sometimes affect the behavior of the ion currents detected by SICM and, especially, make topography measurements difficult. For measuring such charged samples, we propose a novel imaging method that uses a double-barrel nanopipette as an SICM probe. The ion current between the two apertures of the nanopipette desensitizes the surface charge effect on imaging. In this study, metaphase chromosomes of Indian muntjac were imaged by this technique because, owing to their strongly negatively charged surfaces in phosphate-buffered saline, it is difficult to obtain the topography of the chromosomes by the conventional SICM with a single-aperture nanopipette. Using the proposed method with a double-barrel nanopipette, the surfaces of the chromosomes were successfully measured, without any surface charge confounder. Since the detailed imaging of sample topography can be performed in physiological liquid conditions regardless of the sample charge, it is expected to be used for analyzing the high-order structure of chromosomes in relation to their dynamic changes in the cell division.

Ion concentration measurement using synthetic microfluidic papers.

Non-invasive diagnosis on biological liquid samples, such as urine, sweat, saliva, and tears, may allow patients to evaluate their health by themselves. To obtain accurate diagnostic results, target liquid must be precisely sampled. Conventionally, urine sampling using filter paper can be given as an example sampling, but differences in the paper structure can cause variations in sampling volume. This paper describes precise liquid sampling using synthetic microfluidic papers, which are composed of obliquely combined micropillars. Sampling volume accuracy was investigated using different designs and collection methods to determine the optimal design and sample collecting method. The optimized protocol was followed to accurately measure potassium concentration using synthetic microfluidic paper and a commercially available densitometer, which verified the usefulness of the synthetic microfluidic papers for precision sampling.

Actual approaches to the transportation of biological samples at low temperatures.

Taking into account the impact of shipment method of biosamples is necessary for obtaining high-quality biological samples in biobanking and laboratory research. The impact of liquid nitrogen, dry ice and cold accumulators on the quality of biological markers was considered, as well as recommendations to reduce the impact of these methods of shipment. The liquid nitrogen provides the best preservation of samples, however, dry ice is used much more often during their transportation. When transporting certain types of cells using dry ice, there is the way to use CryoStor CS1 and Cell Banker 1 cryoprotectors. The dry ice has a significant effect on both the pH of liquid biological samples and the coagulological parameters of plasma samples. The penetration of CO2 into the sample leads to changes in the parameters of PTT and APPT, as well as to decrease the protein C and fibrinogen level under certain conditions. Serum and plasma samples exposed to dry ice for more than 16 hours should be thawed open at room temperature, or instead of it should be kept at -80 °C for 24 hours to avoid changes in coagulation parameters, The use of cold accumulators is unacceptable for long-term shipment of serum and plasma containing unstable biomarkers because of insufficiently low temperature (increase over time to -25 °C and above). Besides, metal pellets can be used as cold storage batteries at low temperatures (up to -80 ° C), but they are not as effective as dry ice, since it is able to hold the required temperature for much longer.

Extraction of Lipids from Liquid Biological Samples for High-Throughput Lipidomics.

Extraction of the lipid fraction is a key part of acquiring lipidomics data. High-throughput lipidomics, the extraction of samples in 96w plates that are then run on 96 or 384w plates, has particular requirements that mean special development work is needed to fully optimise an extraction method. Several methods have been published as suitable for it. Here, we test those methods using four liquid matrices: milk, human serum, homogenised mouse liver and homogenised mouse heart. In order to determine the difference in performance of the methods as objectively as possible, we used the number of lipid variables identified, the total signal strength and the coefficient of variance to quantify the performance of the methods. This showed that extraction methods with an aqueous component were generally better than those without for these matrices. However, methods without an aqueous fraction in the extraction were efficient for milk samples. Furthermore, a mixture containing a chlorinated solvent (dichloromethane) appears to be better than an ethereal solvent (tert-butyl methyl ether) for extracting lipids. This study suggests that a 3:1:0.005 mixture of dichloromethane, methanol and triethylammonium chloride, with an aqueous wash, is the most efficient of the currently reported methods for high-throughput lipid extraction and analysis. Further work is required to develop non-aqueous extraction methods that are both convenient and applicable to a broad range of sample types.