Nanobiosensorics Laboratory, Centre for Energy Research, Institute of Technical Physics and Materials Science, 1121 Budapest, Hungary.
Nanobiosensorics Laboratory, Centre for Energy Research, Institute of Technical Physics and Materials Science, 1121 Budapest, Hungary.
Adv Colloid Interface Sci. 2022 Oct;308:102727. doi: 10.1016/j.cis.2022.102727. Epub 2022 Jul 6.
In this review we aim to summarize the current state of methods for label-free identification and functional characterization of leukocytes with biosensors and novel single cell techniques. The growing interest in this field is fueled from multiple directions, with the different aspects highlighting benefits of these novel technologies in comparison to classical methods. The advantage of label-free characterization is that labeling the cells might affect their behavior, and therefore lead to a biased description of the investigated biological phenomena. Label-free biosensors can offer the benefit of (i) decreasing processing time and reagent costs, (ii) enable point-of-care diagnostics, and (iii) allow downstream application of the investigated cells. Moreover, (iv) label-free detection allows the monitoring of real-time kinetic processes, opening up new avenues in contrast to traditional structural characterizations. The emphasis in the review will be on techniques on the characterizations of single cells with special attention to surface sensitive technologies. Recent developments highlighted the importance of small cell populations and individual cells both in health and disease. Nonetheless techniques capable of analyzing single cells offer a promising tool for therapeutic approaches where characterization of individual cells is necessary to estimate their clinical therapeutic potential. Most of the approaches discussed here will cover the cellular activation, adhesion as measured on functionalized solid substrates, since this approach offers the most advantages. Analyzing various cells on solid substrates not only allows their individual morphological characterization and therefore a more precise description of their activation, but as well offers an opportunity to design multiplex measurements. With this approach different stimuli can be investigated in parallel and measure cellular avidity to targets, an important aspect of gaining more and more attention recently in characterization of T-cells and antibody effector functions. Finally, novel label-free approaches provide a solution to extracting unlabeled cells for downstream processing (e.g., transcriptome analysis, cloning or the aforementioned clinical potential), where ongoing and potential further applications are discussed.
在这篇综述中，我们旨在总结目前无标记识别和功能表征白细胞的方法，包括生物传感器和新型单细胞技术。由于多个方向的兴趣增长，这些新技术在某些方面相对于传统方法具有优势。无标记特征的优势在于，标记细胞可能会影响其行为，从而导致对所研究生物现象的有偏差描述。无标记生物传感器具有以下优点：（i）减少处理时间和试剂成本；（ii）实现即时诊断；（iii）允许对所研究的细胞进行下游应用。此外，（iv）无标记检测允许实时监测动力学过程，与传统结构表征相比，开辟了新的途径。本综述的重点将放在单细胞特性的技术上，特别关注表面敏感技术。最近的发展强调了小细胞群体和单个细胞在健康和疾病中的重要性。尽管能够分析单细胞的技术提供了一种有前途的工具，但对于需要对单个细胞进行特征分析以评估其临床治疗潜力的治疗方法来说，这种技术仍然是必要的。这里讨论的大多数方法都将涵盖细胞的激活和在功能化固体基底上的黏附，因为这种方法具有最大的优势。在固体基底上分析各种细胞不仅允许对其进行单独的形态学特征描述，从而更精确地描述其激活情况，而且还提供了设计多重测量的机会。通过这种方法，可以同时研究不同的刺激，并测量细胞对靶标的亲和力，这是最近在 T 细胞和抗体效应功能表征中越来越受到关注的一个重要方面。最后，新型无标记方法为提取未标记细胞进行下游处理（例如，转录组分析、克隆或上述临床潜力）提供了一种解决方案，我们还讨论了正在进行的和潜在的进一步应用。
