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Design of an optofluidic sensor for rapid detection of hemolysis
... Another approach using the integration of optical fibers was proposed by Archibong et al. 64 An optical fiber enclosed in a PDMS chamber along with the reflective coating in the bottom was used to detect fHgb with a sensitivity of 4 mg/dL 64 using 50 μL of plasma. Figure 3C shows the conceptual model of the device. ...
... Another interesting optofluidic technique is fabrication of complex 3D-shaped polymer particles in fluid using UV light [3]. Biomedical analysis can be conducted by integration of absorption spectroscopy [4,5] or plasmonic microarrays [6] in a microfluidic system. There are many ways to use optofluidics for control and manipulation of different types of small objects in fluids. ...
It has been demonstrated that optically controlled microcurrents can be used to capture and move around a variety of microscopic objects ranging from cells and nanowires to whole live worms. Here, we present our findings on several new regimes of optofluidic manipulation that can be engineered using careful design of microcurrents. We theoretically optimize these regimes using COMSOL Multiphysics and present three sets of simulations and corresponding optofluidic experiments. In the first regime, we use local fluid heating to create a microcurrent with a symmetric toroid shape capturing particles in the center. In the second regime, the microcurrent shifts and tilts because external fluid flow is introduced into the microfluidic channel. In the third regime, the whole microfluidic channel is tilted, and the resulting microcurrent projects particles in a fan-like fashion. All three configurations provide interesting opportunities to manipulate small particles in fluid droplets and microfluidic channels.
Fluorescent lateral flow immunoassay (LFIA) is one of the most popular strategies for point-of-care testing (POCT), which is capable of rapid screening for disease detection. However, the existing fluorescent LFIA is inapplicable to the hemolytic sample, whose strong absorbance, scattering and autofluorescence in visible region will interfere with the fluorescent signal. To address this issue, we developed a NIR-II LFIA detection platform (termed as NIR-II-LFIA platform) by integrating a portable NIR-II reader and a NIR-II nanoprobes into LFIA. Due to the minimized interaction of NIR-II light with hemolytic sample, this NIR-II LFIA can significantly eliminate the interference of absorbance, scattering and autofluorescence. We further applied this LFIA planform to alpha-fetoprotein (AFP) detection in hemolytic blood sample with high -accuracy and -sensitivity. This strategy exhibits a wide dynamic range of detection by 3 orders of magnitude from 0.80 to 5700 ng/mL. And the detection limit could be as low as 0.24 ng/mL, which is of 100 times lower than the clinical cutoff values (25 ng/mL). Therefore, we believe that this NIR-II-LFIA system can serve as a screening platform for early disease detection, especially in complex matrix samples with severe background interference.
Defined as red blood cell break down and the release of hemoglobin and intracellular contents into the plasma, hemolysis can seriously impact patient care as well as the laboratory's reputation through its affect on test results. Therefore, the European Preanalytical Scientific Committee, in collaboration with the International Federation of Clinical Chemistry Working Group on Patient Safety, have designed a questionnaire to collect data on prevalence and management of hemolytic specimens referred to the clinical laboratories for clinical chemistry testing. This book will help identify the areas where hemolysis occurs most frequently, which can, in turn, guide further analysis about why it is occurring. Once these elements are known, practices and procedures can be implemented to dramatically reduce hemolysis and avoid erroneous laboratory results affecting patient care and increasing laboratory costs.
We propose a new approach to the modular packaging of microfluidic components, in which different functional components are not only fabricated separately but are also designed to be individually removable for the purposes of replacement or subsequent analysis. In this paper, we demonstrate one such component: a stand-alone microfluidic filter that can be custom-fabricated and then connected, disconnected, and replaced on a microfluidic chip as needed. This filter is also designed such that particles captured on the filter can be further analyzed or processed directly on the filter itself - for example, for microscopic examination or cell culturing. The filter is a thin (1 μm) transparent silicon nitride membrane that can be designed and fabricated according to specifications for different applications. This material is suitable for microscale fabrication; filtration of a variety of solutions, including biological samples; and subsequent particle imaging and processing. The porous nature of the thin filter allows for particle separation under relatively low pressures, thus protecting the particles from rupture or membrane damage. We describe two methods for integrating the filter apparatus onto a microfluidic chip such that it can be inserted, removed, and replaced. To demonstrate the utility of this approach, we fabricated custom-designed silicon-based filters, incorporated them onto microfluidic systems then filtered microparticles and live cells from test solutions, and finally removed the filters to image the microparticles and culture the cells directly on the filter membranes.
Administering a wrong drug or a wrong dose can be extremely dangerous and can result in severe adverse effects or even the death of a patient. With human errors being possible, automatic real time identification of a drug and its concentration using technology is a viable option to decrease the chance of incorrect drug administration. As a step toward this goal, we propose a new optical fiber based spectroscopic system that has built-in filtration capabilities and thus can work in real time near patient without additional sample pre-processing. It is designed as a point probe consisting of an optical fiber with a miniature filtering reflector integrated on the interface. In the future it can be inserted into a bag for intravenous therapy (IV therapy) or in a syringe to measure the spectrum of the fluid and to confirm its properties. Additionally, use of microfluidic filtration allows to remove microscopic particles from the sample and thus decreases the noise and increases the sensitivity of spectroscopic measurement. In this study, an optofluidic system was fabricated, and filtration capabilities and measurement of cobalamin (vitamin B12) concentration have been demonstrated.
Hemolysis occurs in many hematologic and non-hematologic diseases. Extracellular hemoglobin (Hb) has been recognized to trigger specific pathophysiologies that are associated with adverse clinical outcomes in patients with hemolysis, such as acute and chronic vascular disease, inflammation, thrombosis and renal impairment. Among the molecular characteristics of extracellular Hb, translocation of the molecule into the extravascular space, oxidative and nitric oxide reactions, hemin release and molecular signaling effects of hemin appear to be the most critical. Limited clinical experience with a plasma-derived haptoglobin product in Japan and more recent preclinical animal studies suggest that the natural Hb and hemin scavenger proteins haptoglobin (Hp) and hemopexin (Hpx) have a strong potential to neutralize the adverse physiologic effects of Hb and hemin. This includes conditions that are as diverse as red blood cell transfusion, sickle cell disease, sepsis and extracorporeal circulation. This perspective reviews the principal mechanisms of Hb and hemin toxicity in different disease states, updates how the natural scavengers efficiently control these toxic moieties, and explores critical issues in the development of human plasma-derived Hp and Hpx as therapeutics for patients with excessive intravascular hemolysis.
Introduction
Hemolysis can be induced in sepsis via various mechanisms, its pathophysiological importance has been demonstrated in experimental sepsis. However, no data on free hemoglobin concentrations in human sepsis are available. In the present study we measured free hemoglobin in patients with severe sepsis as well as in postoperative patients using four methods. It was our aim to determine the potential value of free hemoglobin as a biomarker for diagnosis and outcome of severe sepsis in critical illness.
Methods
Plasma concentration of free hemoglobin was determined in patients with severe sepsis (n = 161) and postoperative patients (n = 136) on day 1 of diagnosis and surgery. For the measurement of free hemoglobin, an enzyme linked immunosorbent assay and three spectrophotometric algorithms were used. Moreover, SAPS II- and SOFA scores as well as procalcitonin concentration and outcome were determined. Kaplan-Meier analysis was performed and odds ratios were determined after classification of free hemoglobin concentrations in a high and low concentration group according to the median. For statistical evaluation the Mann-Whitney test and logistic regression analysis were used.
Results
In non-survivors of severe sepsis, free hemoglobin concentration was twice the concentration compared to survivors. Thirty-day survival of patients, as evidenced by Kaplan-Meier analysis, was markedly lower in patients with high free hemoglobin concentration than in patients with low free hemoglobin concentration. Best discrimination of outcome was achieved with the spectrophotometric method of Harboe (51.3% vs. 86.4% survival, p < 0.001; odds ratio 6.1). Multivariate analysis including free hemoglobin, age, SAPS II- and SOFA-score and procalcitonin demonstrated that free hemoglobin, as determined by all 4 methods, was the best and an independent predictor for death in severe sepsis (p = 0.022 to p < 0.001). Free hemoglobin concentrations were not significantly different in postoperative and septic patients in three of four assays. Thus, free hemoglobin can not be used to diagnose severe sepsis in critical illness.
Conclusions
Free hemoglobin is an important new predictor of survival in severe sepsis.
More than ten years ago, the Institute of Medicine (IOM) reported alarming data on the causes and impact of medical errors in the US, demanding for urgent national eff orts to address this problem. Despite large initiatives to improve patient safety throughout the managed care have growth expo-nentially since the release of "To Err is Human", the outcome has however been much lower than ex-pected. It is still undeniable, however, that the worldwide galloping movement of patient safety, boosted by Governments, national healthcare sys-tems and consumers unions, has catalyzed impor-tant changes in healthcare and laboratory medi-cine as well. Some of these changes have intro-duced important innovations in the current medi-cal and laboratory practice, towards establishment of a foremost culture of quality and safety. The very issue we have to face is natural to the real meaning of "healthcare quality". Quality is gene-rally defi ned as "a high degree or grade of exce-llence", but the translation of this concept in heal-thcare is somehow challenging, since we all know that "quality" means rather diff erent things to dif-ferent people. Someone thinks that getting quali-ty healthcare means seeing the doctor right away, being treated courteously by the doctor's staff , or having the doctor spend a lot of time with him/ her. While all these things are important, the "clini-cal" quality of healthcare is indeed much more pervasive. The Agency for Healthcare Research and Quality (AHRQ) of the U.S. Department of Health & Human Services brings a helpful example to defi -ne healthcare quality. Getting quality health care is like taking a car to the mechanic; the people in the garage can be pleasant and take note of compla-ints, but the most important thing is whether they can be able to fi x the problems and, hopefully, to return the car timely and with no additional mal-functioning. Accordingly, healthcare quality can be seen as receiving the most appropriate care, whilst minimizing the risk of side eff ects and ad-verse events not directly related to the presence of the original disease (e.g., medication errors, wrong site surgery or retained instrument after an opera-tion, wrong drug or wrong route of administration of drugs, adverse drugs reactions, hospital acqui-red infections, etc). In other words, according to the current defi nition of the US Institute of Medici-ne (IOM) healthcare quality is the degree to which health services increase the likelihood of desired health outcomes and are consistent with current professional knowledge.
Hemolysis is still the most common reason for rejecting samples, while reobtaining a new sample is an important problem. The aim of this study was to investigate the effects of hemolysis in different hemolysis levels for mostly used biochemical parameters to prevent unnecessary rejections.
Sixteen healthy volunteers were enrolled in the study. Four hemolysis levels were constituted according to hemoglobin concentrations and they were divided into five groups: Group I: 0-0.10 g/L, Group II:0.10-0.50 g/L, Group III: 0.51-1.00 g/L, Group IV: 1.01-2.50 g/L, Group V: 2.51-4.50 g/L. Lysis was achieved by mechanical trauma.
Hemolysis interference affected lactate dehydrogenase (LD) and aspartate aminotransferase (AST) almost at undetectable hemolysis by visual inspection (plasma hemoglobin < 0.5 g/L). Clinically meaningful variations of potassium and total bilirubin were observed in moderately hemolyzed samples (hemoglobin > 1 g/L). Alanine aminotransferase (ALT), cholesterol, gamma glutamyltransferase (GGT), and inorganic phosphate (P) concentrations were not interfered up to severely hemolyzed levels (hemoglobin: 2.5-4.5 g/L). Albumin, alkaline phosphatase (ALP), amylase, chloride, HDL-cholesterol, creatine kinase (CK), glucose, magnesium, total protein, triglycerides, unsaturated iron binding capacity (UIBC) and uric acid differences were statistically significant, but remained within the CLIA limits.
To avoid preanalytical visual inspection for hemolysis detection, improper sample rejection, and/or rerun because of hemolysis, it is recommended in this study that, routine determination of plasma or serum free hemoglobin concentrations is important. For the analytes interfered with hemolysis, new samples have to be requested.
Background:
In vitro hemolysis, the prevailing cause of preanalytical error in routine laboratory diagnostics, might influence the reliability of several tests, affect the quality of the total testing process and jeopardize patient safety. Although laboratory instrumentation is now routinely equipped with systems capable of automatically testing and eventually correcting for hemolysis interference, to our knowledge there are no reports that have compared the efficiency of different analytical platforms for identifying and classifying specimens with hemolysis.
Methods:
Serum from a healthy volunteer was spiked with varying amounts of hemolyzed blood from the same volunteer, providing a serum free hemoglobin concentration ranging from 0.0 g/L to 2.0 g/L as measured by the reference cyanmethemoglobin assay. The spiked serum samples were shipped to seven separate laboratories and the hemolysis index (HI) was tested in triplicate on the following analytical platforms: Roche Modular System P (n=4) and Integra 400 Plus (n=1), Siemens Dimension RxL (n=3), ADVIA 2400 (n=1) and ADVIA 1800 (n=1), Olympus AU 680 (n=1) and Coulter DXC 800 (n=1).
Results:
Satisfactory agreement of HI results was observed among the various analytical platforms, despite a trend toward overestimation by the ADVIA 2400 and 1800. After normalizing results according to the instrument-specific alert value, discrepancies were considerably reduced. All instruments except for the Dimension RxL gave values normalized to the instrument-specific alert value, <1.0 for the sample with 0.048 g/L free hemoglobin, and >1.0 for the sample with 0.075 g/L free hemoglobin. The results of the four Modular System P tests were also highly reproducible among the different facilities. When evaluating instruments that provided quantitative HI results, the mean intra-assay coefficient of variation (CV) calculated for the triplicate determinations was always between 0.1% and 2.7%.
Conclusions:
The results of this multicenter evaluation confirm that efficiency of different analytical platforms to correctly identify and classify unsuitable samples is satisfactory. However, more effort should be placed on the standardization of reporting HI. All the instruments that we tested provide either quantitative or qualitative results that are essentially comparable, but which should always be compared with the instrument-specific alert values to harmonize their efficiency.
Of eight methods examined for measuring plasma hemoglobin in micromolar concentration, all exhibited acceptable linearity, reproducibility, and concurrence except when specimens were icteric or lipemic or contained methemoglobin or methemalbumin. Measurement of absorbance at 578 nm with an Allen correction permits precise assay of plasma oxyhemoglobin concentration as low as 0.01 g/L (1 mg/dL, 0.16 mumol/L), unaffected by hyperlipidemia or hyperbilirubinemia. Discrepancies between methods occurred in 11.6% of a consecutive series of 50 nonicteric patients' plasma specimens. Examination of absorption spectra is helpful when discrepancies are observed between methods. The presence of methemalbumin or methemoglobin in plasma is not recognized by methods that measure only oxyhemoglobin. Increased ceruloplasmin or beta-carotene does not significantly affect results.
An Allen correction and a polychromatic analysis are about equally effective in minimizing effects of interference by bilirubin and triglyceride turbidity in the direct spectrophotometry of serum hemoglobin: interference from bilirubin is nearly eliminated, that from turbidity substantially decreased. The limit of detectability of hemoglobin is 8 mg/L in the presence of a moderate concentration of bilirubin. A change in hemoglobin concentration as small as 16 mg/L can be detected in serum having a concentration near the upper limit of the reference interval, i.e., at the medical decision level. The polychromatic formula gives concentration estimates approximately 5% greater than those of the Allen correction. The formula for the Allen correction is hemoglobin (mg/L) = 1.68 mA415 - 0.84 mA380 - 0.84 mA450 . That for the polychromatic analysis is hemoglobin (mg/L) = 1.65 mA415 - 0.93 mA380 - 0.73 mA470 .
Enzyme-linked immunosorbent assay (ELISA) is one of the most important technologies for biochemical analysis critical for diagnosis and monitoring of many diseases. Traditional systems for ELISA incubation and reading are expensive and bulky, thus cannot be used at point-of-care or in the field. Here, we propose and demonstrate a new miniature mobile phone based system for ELISA (MELISA). This system can be used to complete all steps of the assay, including incubation and reading. It weighs just 1 pound, can be fabricated at low cost, portable, and can transfer test results via mobile phone. We successfully demonstrated how MELISA can be calibrated for accurate measurements of progesterone and demonstrated successful measurements with the calibrated system.
Real-time detecting phosphate has significant meanings in marine environmental monitoring and forecasting the occurrence of harmful algae blooms. Conventional monitoring instruments are depending on artificial sampling and laboratory analysis. They have various shortcomings in real time applications because of the large equipment size and high production cost, with low target selectivity and time-consuming in obtaining detection results. We proposed an optofluidic miniaturized analysis chip combined with micro-resonators to achieve real-time phosphate detection. The quantitative water-soluble components are controlled by flow rates of the phosphate solution, chromogenic agent A (ascorbic acid solution) and chromogenic agent B (12% ammonium molybdate solution, 80% concentrated sulfuric acid and 8% antimony potassium tartrate solution with volume ratio of them is 80: 18: 2.). Subsequently, an on-chip Fabry–Pérot microcavity is formed with a pair of aligned coated fiber facets. With the help of optical feedback, the absorption of phosphate can be enhanced, which can avoid the disadvantage of macroscale absorption cell in traditional instruments. It can overcome the difficulties of traditional instruments in size, parallel processing of numerous samples and real-time monitoring, etc. The absorption cell length is shortened to 300 µm with the detection limit of 0.1 µmol/L. The time of the detection is shorten from 20 mins to 6 seconds. Predictably, micro sensor based on optofluidic technology will has potentials in the field of marine environmental monitoring.
Preeclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome are pregnancy-related complications with high rates of morbidity and mortality. HELLP syndrome, in particular, can be difficult to diagnose. Recent work suggests that elevated levels of free cell hemoglobin in blood plasma can, as early as the first trimester, potentially serve as a diagnostic biomarker for impending complications. We therefore developed a point-of-care mobile phone–based platform that can quickly characterize a patient's level of hemolysis by measuring the color of of blood plasma. The custom hardware and software are designed to be easy to use. A sample of the whole blood (~10 μL or less) is first collected into a clear capillary tube or microtube, which is then inserted into a low-cost 3D-printed sample holder attached to the phone. A 5–10 min period of quiescence allows for gravitational sedimentation of the red blood cells, leaving a layer of yellowish plasma at the top of the tube. The phone camera then photographs the capillary tube and analyzes the color components of the cell-free plasma layer. The software converts these color values to a concentration of free hemoglobin, based on a built-in calibration curve, and reports the patient's hemolysis level: non-hemolyzed, slightly hemolyzed, mildly hemolyzed, frankly hemolyzed, or grossly hemolyzed.. The accuracy of the method is ~1 mg dL–1. This phone-based point-of-care system provides the potentially life-saving advantage of a turnaround time of about 10 minutes (versus 4+ hours for conventional laboratory analytical methods) and a cost of approximately one dollar USD (assuming you have the phone and the software are already available).
In this paper, we demonstrate the optimization of a capacitively coupled plasma etching for the fabrication of a polysilicon waveguide with smooth sidewalls and low optical loss. A detailed experimental study on the influences of RF plasma power and chamber pressure on the roughness of the sidewalls of waveguides was conducted and waveguides were characterized using a scanning electron microscope. It was demonstrated that optimal combination of pressure (30 mTorr) and power (150 W) resulted in the smoothest sidewalls. The optical losses of the optimized waveguide were 4.1±0.6 dB/cm.
Hemolysis of blood samples creates significant delays in the treatment and disposition of patients in the emergency department. The purpose of this study was to compare the hemolysis rates of coagulation blood samples obtained during insertion of an intravenous (IV) catheter without (group 1) or with (group 2) extension tubing connected to the IV catheter hub. A secondary purpose of this study was to determine whether the investigators could predict whether a coagulation sample was hemolyzed based on visual observation during the specimen withdrawal process.
A prospective, 2-group randomized comparative design was used to determine which method of blood collection for coagulation specimens provided the lowest hemolysis rate. This study was conducted in an urban level I emergency department averaging 58,000 visits per year. The sample consisted of 121 adult ED patients randomly assigned to 1 of the 2 groups. Data collectors were trained in the 2 methods of coagulation sample collection and followed a strict protocol. The clinical laboratory used a standardized color-coded scale to determine hemolysis.
Pearson χ(2) analysis was used to test for differences between all nominal variables. The level of significance for all tests was P < .05. There was no significant difference in hemolysis rates between the groups by use of χ(2) analysis (P = .84). Nurses were significantly more likely to predict that a sample was hemolyzed when it was not and to think that it was not hemolyzed when in fact it was (P < .001).
High hemolysis rates occurred equally when coagulation blood samples were drawn via a peripheral IV catheter either at the hub or through extension tubing. Emergency nurse investigators could not accurately predict by visualization whether a coagulation sample was hemolyzed at the time of blood withdrawal. Venipuncture as the preferred method of blood draw is an industry recommendation. This method has been shown in prior experimental studies to reduce hemolysis rates to less than 4%. Therefore, if hemolysis rates are a concern, one should consider obtaining blood whenever possible through a venipuncture rather than through an IV catheter. Replication studies are needed to determine whether the findings of this study can be generalized to the larger population.
Storage of red cells causes a progressive increase in hemolysis. In spite of the use of additive solutions for storage and filters for leucoreduction, some amount of hemolysis is still inevitable. The extent of hemolysis, however, should not exceed the permissible threshold for hemolysis even on the 42(nd) day of storage.
Eighty units of packed red cells, 40 stored in SAGM post leucoreduction and 40 in ADSOL without leucoreduction filters, were evaluated for plasma hemoglobin by HemoCue Plasma Hemoglobin analyzer on the day of collection and on the 7(th), 14(th), 21(st), 28(th), 35(th) and 42(nd) days thereafter. The hemoglobin and hematocrit were also noted for all these units by the Beckman and Coulter analyzer. Percentage hemolysis was then calculated.
Hemolysis progressively increased with the storage period in all the stored red cell units (SAGM as well ADSOL). However, on day 42(nd) of storage, free hemoglobin in all the red cell units was within the permissible level (which is 0.8% according to the Council of Europe guidelines and 1% as per the US FDA guidelines). The mean percentage hemolysis was slightly higher in the SAGM-containing bags with an integral leucoreduction filter as compared to the bags containing ADSOL. However this difference was marginal and not statistically significant.
Hemolysis of the red cells increases with storage. However, maximum hemolysis does not exceed the permissible limits at any time thereby indicating the effect of optimum processing and storage conditions on red cell hemolysis.
- E Archibong
E. Archibong, et al.
Sensors & Actuators: B. Chemical 288 (2019) 274-278
Evaluation of red cell hemolysis in packed red cells during processing and storage
- Makroo