A New Method for Ultra-sensitive P24 Antigen Assay Based on Near-infrared Fluorescent Microsphere Immunochromatography

Objective To develop a rapid,highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.Methods First,we prepared a lateral flow assay test strip,and labeled the detection antibody using a fluorescent microsphere.Second,we optimized the antibody labeling conditions.Third,we optimized the detection conditions.Fourth,we created a working curve.Fifth,we conducted a methodological assessment of the established fluorescent microsphere immunochromatography method.Sixty-six clinical samples were tested,and we compared the established fluorescent microsphere immunochromatography with the quantitative ELISA method.Results According to the working curve,the detection limit of the method is 3.4 pg/mL,and the detection range is 3.4 pg/mL to 10 ng/mL.The average intra-assay recovery was 99.6%,and the Coefficient of Variation(CV)was 5.4%–8.6%;the average inter-assay recovery was 97.3%,and the CV was 8.5%–11%.The detection rate of fluorescent microsphere immunochromatography was higher than ELISA method,and had a good correlation with ELISA.Conclusion The P24 antigen quantitative detection method based on near-infrared fluorescent microsphere immunochromatography has the advantages of rapid detection,high sensitivity,and wide detection range;thus,it is suitable for early clinical diagnosis and continuous monitoring of AIDS.

Histochemistry of microinfarcts in the mouse brain after injection of fluorescent microspheres into the common carotid artery

The mouse model of multiple cerebral infarctions,established by injecting fluorescent microspheres into the common carotid artery,is a recent development in animal models of cerebral ischemia.To investigate its effectiveness,mouse models of cerebral infarction were created by injecting fluorescent microspheres,45–53μm in diameter,into the common carotid artery.Six hours after modeling,fluorescent microspheres were observed directly through a fluorescence stereomicroscope,both on the brain surface and in brain sections.Changes in blood vessels,neurons and glial cells associated with microinfarcts were examined using fluorescence histochemistry and immunohistochemistry.The microspheres were distributed mainly in the cerebral cortex,striatum and hippocampus ipsilateral to the side of injection.Microinfarcts were found in the brain regions where the fluorescent microspheres were present.Here the lodged microspheres induced vascular and neuronal injury and the activation of astroglia and microglia.These histopathological changes indicate that this animal model of multiple cerebral infarctions effectively simulates the changes of various cell types observed in multifocal microinfarcts.This model is an effective,additional tool to study the pathogenesis of ischemic stroke and could be used to evaluate therapeutic interventions.This study was approved by the Animal Ethics Committee of the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences(approval No.D2021-03-16-1)on March 16,2021.

Preparation and performance of fluorescent polyacrylamide microspheres as a profile control and tracer agent

Polyacrylamide microspheres have been suc- cessfully used to reduce water production in reservoirs, but it is impossible to distinguish polyacrylamide microspheres from polyacrylamide that is used to enhance oil recovery and is already present in production fluids. In order to detect polyacrylamide microspheres in the reservoir pro- duced fluid, fluorescent polyacrylamide microspheres P（AM-BA-AMCO）, which fluoresce under ultraviolet irradiation, were synthesized via an inverse suspension polymerization. In order to keep the particle size distribu- tion in a narrow range, the synthesis conditions of the polymerization were studied, including the stirring speed and the concentrations of initiator, NaaCO3, and dispersant. The bonding characteristics of microspheres were deter- mined by Fourier transform infrared spectroscopy. The surface morphology of these microspheres was observed under ultraviolet irradiation with an inverse fluorescence microscope. A laboratory evaluation test showed that the fluorescent polymer microspheres had good water swelling capability, thus they had the ability to plug and migrate in a sand pack. The plugging rate was 99.8 % and the residual resistance coefficient was 800 after microsphere treatment in the sand pack. Furthermore, the fluorescent microspheres and their fragments were accurately detected under ultra- violet irradiation in the produced fluid, even though theyhad experienced extrusion and deformation in the sand pack.

Chondroitin Sulfate Fluorescence Biosensor Based on Graphere Quantum Dots Aggregating on CMC/CS Polyelectrolyte Microspheres

Here, we report an efficient fluorescence biosensor for chondroitin sulfate（CHS） based on polyelectrolyte microspheres of carboxymethyl cellulose（CMC） and chitosan（CS） composites inducing the aggregation of graphene quantum dots（GQDs）, calling CMC/CS-GQDs. The polyelectrolyte microspheres（CMC/CS microspheres） were fabricated by using anioniccationic electrostatic attraction between CMC and CS by high voltage electrostatic spray technology. The aggregating process of GQDs was based on the anionic-cationic electrostatic attraction as well. After combing with the polyelectrolyte microspheres, the fluorescence of GQDs disappeared. CHS, which widely consists in the cell surface of human beings and animals, carries a large number of negative charges on the surface. The addition of CHS enabled CHS and GQDs to compete with each other to composite with the CMC/CS microshpheres. As a result of the higher surface charge density of CHS, CMC/CS-CHS formed accompanied by the release of GQDs, and the fluorescence of the system recovered. The CHS content was detected by analyzing the system＇s fluorescence recovery, which suggested that the obtained fluorescence biosensor can accurately detect the concentration of CHS. The test results showed that the linear range of the fluorescence recovery for this biosensor with respect to CHS was 0~12.00 mg/mL, and the detection limit was 10-8 M. Besides, to test the stability of the biosensor, the CMC/CS-GQDs micropsheres persisted for one month, with a low fluorescence quenching of 9.48%. These results suggested that CMC/CS-GQDs can be utilized as efficient fluorescence biosensor for the detection of CHS. Moreover, the detection method was simple and efficient, and could be widely popularized.

Manipulable multipurpose nanothermometers based on a fluorescent hybrid glass fiber microsphere cavity

Fluorescent nanothermometers for remote temperature measurement at the micro/nanoscale have stimulated growing efforts in developing efficient temperature-responsive materials and detection procedures.However,the efficient collection and transmission of optical signals have been a tremendous challenge for practical applications of these nanothermometers.Herein,we design an all-fiberized thermometry based on a fiber-coupled microsphere cavity coated with thermo-sensitive NaYF_(4)∶20%Yb^(3+);2%Er^(3+)@NaYF_(4)nanocrystals(NCs),allowing for spatial temperature sensing with resolution down to the few-micrometer scale.In our design,the microsphere efficiently excites the NCs and collects their upconversion emissions,and the use of a fiber splitter coupled with the microsphere allows for lossless routing of excitation and emitted light.We demonstrate the use of this all-fiber temperature sensor in diverse environments,especially in strongly acidic and alkaline conditions.Leveraging the high flexibility of commercial silica fiber,this all-fiber temperature ensor was employed for stable fixed-point real-time temperature measurement and multipurpose temperature recording/mapping in opaque environments,microscale areas,various solutions,and complicated bent structures.Thus,the demonstrated design could have strong implications for the practical use of nanothermometers in various possible scenarios,especially monitoring temperatures in diverse physiological settings.

Comparison of the emission wavelengths by a single fluorescent dye on in vivo 3-photon imaging of mouse brains

Multiphoton microscopy(MPM)is a powerful imaging technology for brain research.The imaging depth in MPM is partly determined by emission wavelength of fluorescent labels.It has been demonstrated that a longer emission wavelength is favorable for signal detection as imaging depth increases.However,there has been no comparison with near-infrared(NIR)emission.In order to quantitatively analyze the effect of emission wavelength on 3-photon imaging of mouse brains in vivo,we utilize the same excitation wavelength to excite a single fluorescent dye and simultaneously collect NIR and orange-red emission fluorescence at 828 nm and 620 nm,respectively.Both experimental and simulation results show that as the imaging depth increases,NIR emission decays less than orange-red fluorescent emission.These results show that it is preferable to shift the emission wavelength to NIR to enable more e±cient signal collection deep in the brain.

Synthesis of Micron-size Functional Polystyrene Fluorescent Micro- spheres and their Adsorbability to Human Serum Albumin

Polystyrene microspheres with sulfo- or aldehyde- surface were synthesized through dispersion polymerization. Functional polystyrene fluorescent microspheres were prepared by the way of adding 2, 5-diphenyloxazole (PPO) into the reaction system directly and dying the blank microspheres in the ethanol solution of PPO. The influence of preparing matters on the encapsulating rate of PPO, and the influence of functional groups on the adsorbability to human serum albumin (HSA) were investigated.

Studies on Monodispersed Microspheres of Zinc Sulfide Doped with Mn^(2+)

In this paper, zinc acetate, manganese acetate and thiacetamide are used as raw materials to successfully synthesize monodispersed ZnS：Mn^2＋ microspheres by using hydrothermal method and taking P123 surfactant as a template. The products were characterized by XRD, STEM, FT-IR and N2 adsorption-desorption. And the results show that the diameter of this microsphere is 1.0 μm or so, which is larger than that of ZnS microsphere without Mn^2＋ doping, and it has monodispersion, smooth surface and uniform size, The doping of Mn^2＋ does not obviously change the structure of monodispersed ZnS microsphere. The photoluminescence peak lies in a wide band ranging from 450 to 650 nm, and the microspheres emit orange light; with the increase of Mn^2＋ concentration, fluorescence intensity of ZnS：Mn^2＋ microsphere changes, and when the mole ratio of Mn^2＋：Zn^2＋ is 0.3：1, the fluorescence intensity is the strongest.

Liver microcirculation after hepatic artery embolization with degradable starch microspheres in vivo

AIM： To observe the dynamic changes of liver microcirculation in vivo after arterial embolization with degradable starch microspheres （DSM）. METHODS： DSM were injected into the proper hepatic artery through a silastic tube inserted retrogradely in gastroduodenal artery （GDA） of SD rats. Fluorescent microscopy was used to evaluate the dynamic changes of blood flow through the terminal portal venules （TPVs）, sinusoids and terminal hepatic venules （THVs）. The movements of DSM debris were also recorded. Six hours after injection of DSM, percentages of THVs with completely stagnant blood flow were recorded. RESULTS： Two phases of blood flow change were recorded. In phase one： after intra-arterial injection of DSM, slow or stagnant blood flow was immediately recorded in TPVs, sinusoids and THVs. This change was reversible, and blood flow resumed completely. In phase two： after phase one, blood flow in TPVs changed again and three patterns of blood flow were recorded. Six hours after DSM injection, 36.9% ± 9.2% of THVs were found with completely stagnant blood flow. CONCLUSION： DSM can stop the microcirculatory blood flow in some areas of liver parenchyma. Liver parenchyma supplied by arteries with larger A-P shunt is considered at a higher risk of total microcirculatory blood stagnation after injection of DSM through hepatic artery.

Ratiometric fluorescence probe for accurate detection of Concanavalin A by coupling fluorescent microsphere with boric acid functionalized carbon dots

Accurate and sensitive strategies for Concanavalin A(Con A)sensing are conducive to the better cognition of various important biological and physiological processes.Here,by designing dextran-functionalized fluorescent microspheres(DxFMs)and boric acid-modified carbon dots(BCDs)as recognition unit and built-in signal reference respectively,a ratiometric fluorescent detection platform was proposed for Con A detection with high reliability.In this protocol,the BCDs/DxFMs precipitation was formed due to the covalent interactions between cis-diol of DxFMs and boronic acid groups of BCDs,thus only fluorescence of BCDs could be detected in the supernatant.When Con A was presented,it could bind to DxFMs through its carbohydrate recognition ability and suppress the subsequent assembly between DxFMs and BCDs,leading to the simultaneous capture of DxFMs and BCDs fluorescence in the supernatant.Since the BCDs content was superfluous,their fluorescence intensities were basically constant in all cases.Based on the unchanged BCDs fluorescence signal and target-dependent DxFMs fluorescence signal in supernatant,the ratiometric detection of Con A was realized.Under optimized conditions,this ratiometric fluorescent platform displayed a linear detection range from 0.125μg/mL to 12.5μg/mL with a detection limit of 0.089μg/mL.Moreover,satisfied analytical outcomes for Con A detection in serum samples were obtained,manifesting huge application potential of this ratiometric fluorescent platform in clinical diagnosis.

4种含笑叶片叶绿素荧光参数Fv/Fm特性的比较

对深山含笑、阔瓣含笑、金叶含笑和峨眉含笑幼树叶绿素荧光参数Fv/Fm在不同季节的日变化及其与光照(PAR)、气温(Ta)和大气相对湿度(RH)的相关性进行了测定和分析。结果表明,4种含笑幼树叶绿素荧光参数Fv/Fm在春季日变化小,各个时刻的测定值均大于0.75,荧光参数Fv/Fm在春季与其PAR、Ta和RH均无显著相关性。4种含笑叶绿素荧光参数Fv/Fm在夏季的日变化均呈先降后升的趋势,最低值(<0.75)出现在12:00～14:00之间。深山含笑和阔瓣含笑叶绿素荧光参数Fv/Fm与夏季的Ta和RH均分别呈显著负相关和显著正相关,但金叶含笑夏季叶绿素荧光参数Fv/Fm与PAR和Ta呈显著负相关。从叶绿素荧光参数Fv/Fm的日变化趋势和测定值看,深山含笑、阔瓣含笑和金叶含笑的光合机构在夏季12:00～14:00均受到可逆性胁迫影响。峨眉含笑夏季叶绿素荧光参数Fv/Fm与Ta和RH分别呈极显著负相关和极显著正相关,且Fv/Fm的测定值在一天中各个时刻均低于0.75,表明其光合机构在夏季受到不可逆性胁迫影响。峨眉含笑叶片叶绿素荧光参数Fv/Fm在秋季日变化呈先降后升的趋势,最低值(0.72)出现在14:00,且与RH呈显著正相关,其它3种含笑的Fv/Fm在秋季的日变化小,各个时刻的测定值均在0.75以上,且与其PAR、Ta和RH无显著相关性。4种含笑叶绿素荧光参数Fv/Fm在冬季的日变化均呈先升后降的趋势,且均与Ta呈显著正相关。从叶绿素荧光参数Fv/Fm的日变化趋势和测定值看,4种含笑的光合机构在冬季只受到了可逆性胁迫影响。

fMLP、PMA、LTB_4对大鼠吞噬细胞能力的影响

目的观测fMLP、PMA、LTB4对大鼠吞噬细胞能力的影响。方法通过密度梯度离心法分离大鼠腹腔吞噬细胞,对分离后得到的巨噬细胞及中性粒细胞使用fMLP、PMA、LTB4诱导后,通过荧光显微镜观察其对荧光微球的吞噬能力,进行相关判断。结果静息状态下,大鼠腹腔巨噬细胞对荧光微球的吞噬能力显著大于中性粒细胞;经fMLP、PMA、LTB4诱导后,巨噬细胞及中性粒细胞对荧光微球的吞噬能力增强。结论大鼠腹腔中性粒细胞吞噬能力小于巨噬细胞;fMLP、PMA、LTB4可致大鼠中性粒细胞、巨噬细胞的的吞噬能力提高。

Versatile core-shell magnetic fluorescent mesoporous microspheres for multilevel latent fingerprints magneto-optic information recognition

Latent fingerprints are extremely vital for personal identification and criminalinvestigation,and potential information recognition techniques have been widelyused in the fields of information and communication electronics.Although physicalpowder dusting methods have been frequently employed to develop latent fingerprintsand most of them are carried out by using single component powders ofmicron-sized fluorescent particles,magnetic powders,or metal particles,there isstill an enormous challenge in producing high-resolution image of latent fingerprintsat different backgrounds or substrates.Herein,a novel and effectivenanoimpregnation method is developed to synthesize bifunctional magnetic fluorescentmesoporous microspheres for latent fingerprints visualization by growthof mesoporous silica(mesoSiO_(2))on magical Fe_(3)O_(4) core and then deposition offluorescent YVO4:Eu^(3+)nanoparticles in the mesopores.The obtainedFe_(3)O_(4)@mesoSiO_(2)@YVO4:Eu^(3+)microspheres possess spatially isolated magneticcore and fluorescent shell which were insulated by mesoporous silica layer.Dueto their small particle size of submicrometer scale,high magnetization and lowmagnetic remanence as well as the combined magnetic and fluorescent properties,the microspheres show superior performance in visual latent fingerprint recognitionwith high contrast,high anti-interference,and sensitivity as well as goodretention on multifarious substrates regardless of surface permeability,roughness,refraction,colorfulness,and background fluorescence interference,and it makesthem ideal candidates for practical application in fingerprint visualization andeven magneto-optic information storage.

Preparation and Properties of Magnetic-fluorescent Microporous Polymer Microspheres

Microporous microspheres can be used as functional nanomaterial carriers for their microporous structure and higher specific surface area. In this study, magnetic fluorescent polymer microspheres were prepared by incorporating Fe304 nanoparticles and CdSe/ZnS quantum dots（QDs） into hyper-crosslinked microporous polymer micro- spheres（HCMPs） via the in situ coprecipitation method and swelling-diffusion. The HCMPs predominantly have mi- cropores, and their specific surface area is as high as 703.4 m2/g. The magnetic-fluorescent microspheres maintain the superparamagnetic behavior of Fe304, and the saturation magnetization reaches 38.6 A.m2/kg. Moreover, the composite microspheres exhibit an intense emission peak at 530 nm and achieve good fluorescence.

可溶性fms样酪氨酸激酶-1荧光免疫层析法检测的临床性能评估

目的评估荧光免疫层析法的可溶性fms样酪氨酸激酶-1(sFlt-1)检测试剂盒的临床性能。方法参照美国临床实验室标准化协会(CLSI)EP6-A、EP7-A、EP15-A文件对试剂盒的精密度、线性、方法学比对等临床性能进行评估。结果试剂盒的高浓度和低浓度样本的变异系数(CV)分别为3.63%和5.92%;线性为40~85 000 pg/mL;与电化学发光法试剂盒对比结果显示,相关性r为0.985,总体符合率为96.71%。结论荧光免疫层析法的sFlt-1检测试剂盒的精密度、线性、方法学比对等临床性能评价较好,满足临床应用的需求。

采出水中核壳型荧光微球浓度的检测方法

聚合物微球调驱是改善水驱效果的主要技术之一。微球在地层中的运移以及能否在采出水中有效检出会直接影响调驱效果。因此,将荧光碳点引入微球调驱剂中,起到示踪的作用。荧光核壳微球调驱剂由含荧光碳点的核心微球溶液和壳层水溶液混合吸附而成。为了降低油水分离后采出水中的杂质对核壳荧光微球有效检出的干扰,首先对荧光微球的浓度与荧光强度进行线性拟合,验证该方法的可行性;然后用硅胶对地层采出水进行吸附,通过对比采出水吸附前后的荧光发射光谱,验证硅胶吸附的实用性;最后用硅胶对采出水配制的荧光微球进行吸附,绘制荧光强度和微球浓度的标准曲线。结果表明,在激发波长为347 nm的条件下,荧光微球的质量浓度与445 nm发射波长处的荧光强度具有良好的线性关系,相关判定系数(R^(2))为0.9870。经硅胶处理后,水驱采出水的荧光发射强度显著降低,硅胶吸附能有效去除采出水中的杂质。在激发波长为347 nm、荧光光谱仪狭缝为10~20 nm、微球质量浓度为1~1200 mg/L时,荧光核壳微球水分散液的质量浓度(x)与462 nm处的荧光发射峰值(y)呈正比线性关系,拟合方程为y=2497.1042+3.1847x,R^(2)为0.9972,置信度较高。荧光强度与核壳微球浓度的线性阶段可满足现场检测要求。该方法可为类似油藏荧光微球含量的定量检测提供借鉴。

基于P32蛋白的山羊痘病毒抗体荧光微球检测方法的建立

为建立一种山羊痘病毒(GTPV)抗体快速检测方法,试验将携带GTPV结构蛋白P32基因的重组质粒pET28a-P32转化至大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导后获得了重组蛋白,纯化后经Western blot鉴定表明该蛋白具有良好的特异性和反应原性。以荧光微球为标记材料对P32蛋白进行标记,通过优化反应条件制备了快速检测GTPV抗体的荧光微球免疫层析试纸条,并对其性能进行评价。结果:该试纸条与山羊副流感病毒3型(CPIV3)、小反刍兽疫病毒(PPRV)、羊口疮病毒(ORFV)的阳性血清均无交叉反应;阳性血清稀释至800倍仍能检出阳性,敏感性较高;批内变异系数小于10%,批间变异系数小于15%,稳定性较好;与ELISA检测方法对比的总符合率为92.5%。综上,本试验建立的荧光微球试纸条可用于GTPV抗体的快速检测和流行病学调查。

溶胀法制备荧光微球研究进展

溶胀法作为一种可调控性强、操作简单、实用性高的制备荧光微球的方法而被广泛应用。本文综述了利用溶胀法制备荧光微球的研究进展,阐述了溶胀法制备荧光微球的基本原理和步骤。进一步综述了溶胀过程中影响微球性能的常见因素,例如聚合物基质、荧光物质、溶胀剂的种类等,总结了溶胀法调控优化荧光微球的改进策略,对溶胀法制备荧光微球的应用前景进行了展望。