流式细胞术最新进展及临床应用

近年来,流式细胞术作为临床检验中无可替代的重要技术之一,在定量方式、检测内容、多参数分析、细胞成像、体内监测及数据分析等方面均取得了许多突破,应用日趋广泛,前景可观。本文将介绍流式细胞术在定量流式细胞术、多色流式细胞术、磷酸化流式细胞术、质谱流式细胞术、成像流式细胞术及体内流式细胞术方面的最新进展及临床应用。     

流式细胞术的原理及临床应用

流式细胞术 (flowcytometry)是 2 0世纪 70年代发展起来的对单细胞定量分析的一种新技术 ,它借鉴了荧光标记技术、激光技术、单抗技术和计算机技术 ,具有极高的检测速度与统计精确性 ,而且从单一细胞可以同时测得多个参数 ,为生物医学与临床检验提供了一个全新视角和强有力的手段。目前 ,随着单克隆抗体技术的发展 ,流式细胞仪检测技术已经广泛使用在基础研究和临床实践的各个方面 ,在细胞生物学、肿瘤学、血液学、免疫学、药理学、遗传学及临床检验学等领域内发挥着重要作用[1] 。一、流式细胞仪检测的原理[2 ,3 ]流式细胞仪主要包括以下几…     

医学免疫学实验教学改革与探索北大核心CSCD

为调动学生对医学免疫学实验课程学习的积极性，提高实验教学效果，通过精简传统免疫学实验项目、增加流式细胞术实验、改进吞噬细胞功能检测方法、进行生物安全知识培训、改进考核模式等方面进行教学改革，使学生更加重视免疫学实验课程的学习，加强学生的生物安全意识，提高不同学生之间学习成绩的区分度。     

多参数流式细胞术检测急性髓系白血病微小残留病的研究

目的探讨多参数流式细胞术检测急性髓系白血病微小残留病的研究进展。方法采用多参数流式细胞术检测微小残留病，其优势在于MFC通过对白血病相关免疫表型检测，比RQ-PCR技术更广泛、直观地对残留白血病细胞进行准确定量。目前MRD已经可用于大于90%以上的急性髓系白血病的免疫表型检测。结果MFC在急性髓系白血病MRD的检测中是最有前景的一种方法。结论 MFC检测MRD对急性髓系白血病的治疗具有重要的指导意义。但目前MRD的检测由于不同的实验室所采用的化疗方案不统一，故在抗体组合方案、阈值的界定、检测时间点的有所不同，故建立MFC检测MRD的标准化是今后发展方向。     

流式荧光微球技术的应用现状及进展

流式荧光微球技术是一种新的高通量检测技术,该技术是以荧光编码微球作为传统免疫学测定、亲和力测定及DNA杂交测定的固相载体,通过流式细胞仪进行检测的新的可用于高通量筛查的多路测定法.通过使用不同荧光编码的微米大小的聚合微球,流式荧光微球技术可以同时分析复杂样品中的多种分析物.每一种微球表面被修饰使其与相应的待测抗原、抗体或寡核苷酸反应,将不同微球混合即可进行多重复合测定.该技术结合了荧光编码微球的特异性与流式细胞仪的高度敏感性.作为一个技术分析平台,流式荧光微球技术在科研及临床应用中将具有重要的潜力.就其应用现状及进展作一综述.     

基于流式细胞术评价单核细胞介导的抗体依赖性细胞介导的细胞毒效应的方法学建立和应用

目的：建立基于流式细胞术的评价单核细胞介导的抗体依赖性细胞介导的细胞毒效应的检测方法。方法 PKH26和CFSE染色的P815细胞为靶细胞，与P815特异性抗体孵育形成抗原抗体复合物，加入外周血单个核细胞作为效应细胞，共同孵育后流式细胞术检测 CD3－CD14＋PKH26＋CFSE－细胞群的百分比，并确定最佳效靶比及效应细胞和靶细胞的孵育时间。运用上述方法对23例HCV慢性感染者和22例健康人的单核细胞介导的抗体依赖性细胞毒作用（ antibody depend-ent cellular cytotoxicity ， ADCC）进行比较分析。结果可通过流式细胞技术检测CD3－CD14＋PKH26＋CFSE－细胞群来评价单核细胞介导的ADCC效应，最佳效靶比为10∶1，最佳杀伤孵育时间为4 h。慢性HCV感染者单核细胞介导的ADCC效应较健康对照明显降低（ P＝0．009）。结论本研究建立了基于流式细胞术的单核细胞介导的ADCC效应的检测方法，为病毒感染及药物研发中免疫学评价提供快速、敏感、安全的检测手段。     

研究生组织学技术教学的探索与实践

《组织学技术》作为一门实践性很强的实验科学,是研究生教学的基础课程,也是实现人才培养目标的重要教学环节。我们从转变教学观念、拓宽教学内容及改革教学方法、加强教材建设、加强实践环节、强化考核手段等方面进行了探索,取得了良好效果。     

液相芯片在生物医学中的研究进展

液相芯片技术是以不同荧光编码微球作为生物探针的载体,在悬浮液态体系中进行生物分子间的反应,以流式细胞术作为光学检测手段的生物芯片技术.介绍了液相芯片在自身免疫病诊断、细胞因子、传染性疾病、内分泌疾病、神经系统疾病及肿瘤标志物的检测等生物医学领域内的应用,最后对液相芯片的应用前景进行了展望.液相芯片技术由于具有高灵敏度、...     

胎儿骨髓间充质干细胞的分离培养及表面标志的检测

目的 建立胎儿骨髓间充质干细胞（mesenchymal stem cells,MSC)的分离培养方法，并对其表面标志进行检测。方法 采用体外细胞培养技术，分离培养胎儿骨髓MSC，用流式细胞术对其进行鉴定，并研究其增殖及生长特征。结果 流式细胞术证明，MSC具有间质细胞的特征。原代及传代培养显示，胎儿骨髓MSC具有活跃增殖的能力。结论 胎儿骨髓MSC作为组织工程重建的种子细胞具有较强的可行性。     

研究生组织学技术教学改革尝试

组织学技术作为研究生教学课程,对研究生进一步掌握机体形态结构和生理机能之间的关系有非常重要的作用.作为一门应用性较强、发展较快的技术,在教学过程中需要不断地补充研究方法的最新进展,这在一定程度上增加了教学的难度.因此在教学过程中如何提高教学效果,使研究生既掌握基本的理论方法,又能灵活地运用到将来的科研工作中,是这门课程最关键的问题.我们在近年的教学实践过程中进行了以下几方面的改革,取得了良好的效果.     

Tolling for autoimmunity-prime time for 7

The reason certain self-antigens are consistently targeted by autoantibodies may be because they are self-adjuvants. Two papers in this issue of Immunity provide important insights into the contribution of Toll-like receptors in systemic autoimmune disease (Berland et al., 2006; Christensen et al., 2006).     

Receptor editing for better or for worse

Receptor editing has emerged from its original identification as a minor secondary mechanism of B cell tolerance to be considered as a dominant mechanism by which autoreactive immature B cells are rendered tolerant. Clonal deletion, previously regarded as the major mechanism of central B cell tolerance, has been shown by recent studies to operate secondarily and only when receptor editing is unable to provide a non-autoreactive specificity. Receptor editing has also been shown to operate during the development of wild-type B lymphocytes, and ongoing investigations demonstrate the influence of particular signaling molecules in the induction and/or inhibition of receptor editing. Together, these studies begin to map the signaling pathways that regulate receptor editing in autoreactive and non-autoreactive immature B cells.     

Prospects for peptide-based immunotherapy for dog allergy

PURPOSE OF REVIEW: Tolerance to ubiquitous environmental substances, allergens, can be accomplished with specific immunotherapy. Conducting therapy with allergen peptides helps to avoid immediate allergic reactions. Dogs are a source of important indoor allergens, which necessitates the development of effective modes of therapy against the allergy they cause. RECENT FINDINGS: The human T-cell epitopes of the major dog allergen Can f 1 were determined recently. They were found to be distributed in seven epitope regions along the molecule. For the peptide immunotherapy of dog allergy, using a pool of seven peptides, one from each of the epitope regions of Can f 1, seems at present to be the best approach. As Can f 1 is not as immunodominant as the main allergens of some other mammals, it remains to be seen whether peptides from other dog allergens should be included in the pool. SUMMARY: The use of a pool of seven peptides from the T-cell epitope regions of Can f 1 is likely to be feasible for treating dog allergy in a great majority of Caucasian populations. In the future, patient-tailored preparations of variants of the T-cell epitope-containing peptides may offer a way to enhance the efficacy of peptide-based immunotherapy.     

Materials for phantoms for terahertz pulsed imaging

Phantoms are commonly used in medical imaging for quality assurance, calibration, research and teaching. They may include test patterns or simulations of organs, but in either case a tissue substitute medium is an important component of the phantom. The aim of this work was to identify materials suitable for use as tissue substitutes for the relatively new medical imaging modality terahertz pulsed imaging. Samples of different concentrations of the candidate materials TX151 and napthol green dye were prepared, and measurements made of the frequency-dependent absorption coefficient (0.5 to 1.5 THz) and refractive index (0.5 to 1.0 THz). These results were compared qualitatively with measurements made in a similar way on samples of excised human tissue (skin, adipose tissue and striated muscle). Both materials would be suitable for phantoms where the dominant mechanism to be simulated is absorption (approximately 100 cm(-1) at 1 THz) and where simulation of the strength of reflections from boundaries is not important; for example, test patterns for spatial resolution measurements. Only TX151 had a frequency-dependent refractive index close to that of tissue, and could therefore be used to simulate the layered structure of skin, the complexity of microvasculature or to investigate frequency-dependent interference effects that have been noted in terahertz images.     

Design methodology for aids for the disabled

Although many aids for the disabled appear to be quite simple, their design is subject to many difficulties. These problems arise because of the intimate relationship between the equipment and the human body. Equipment design is therefore affected by many biological variables that are difficult to define at the beginning of the design exercise and many will not even be obvious until a device is tried out with a user. Standard design methodologies rely on all the variables being defined in a thorough specification at the start of the design process and therefore often lead to ineffective devices or very long development programmes. An alternative methodology is presented which overcomes the problems by separating the user interface aspects of the design from the supporting features. The user interface aspects are then allowed to evolve in conjunction with tests with potential users and finally integrated with the supporting features once a satisfactory solution has been found. Much more effective devices result with much shorter development effort.