Phillips Classic Laser and Nanomedicine Laboratories, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Methods. 2012 Jul;57(3):280-96. doi: 10.1016/j.ymeth.2012.06.009. Epub 2012 Jun 26.
Conventional flow cytometry using scattering and fluorescent detection methods has been a fundamental tool of biological discoveries for many years. Invasive extraction of cells from a living organism, however, may lead to changes in cell properties and prevents the long-term study of cells in their native environment. Here, we summarize recent advances of new generation flow cytometry for in vivo noninvasive label-free or targeted detection of cells in blood, lymph, bone, cerebral and plant vasculatures using photoacoustic (PA) detection techniques, multispectral high-pulse-repetition-rate lasers, tunable ultrasharp (up to 0.8 nm) rainbow plasmonic nanoprobes, positive and negative PA contrasts, in vivo magnetic enrichment, time-of-flight cell velocity measurement, PA spectral analysis, and integration of PA, photothermal (PT), fluorescent, and Raman methods. Unique applications of this tool are reviewed with a focus on ultrasensitive detection of normal blood cells at different functional states (e.g., apoptotic and necrotic) and rare abnormal cells including circulating tumor cells (CTCs), cancer stem cells, pathogens, clots, sickle cells as well as pharmokinetics of nanoparticles, dyes, microbubbles and drug nanocarriers. Using this tool we discovered that palpation, biopsy, or surgery can enhance CTC release from primary tumors, increasing the risk of metastasis. The novel fluctuation flow cytometry provided the opportunity for the dynamic study of blood rheology including red blood cell aggregation and clot formation in different medical conditions (e.g., blood disorders, cancer, or surgery). Theranostics, as a combination of PA diagnosis and PT nanobubble-amplified multiplex therapy, was used for eradication of CTCs, purging of infected blood, and thrombolysis of clots using PA guidance to control therapy efficiency. In vivo flow cytometry using a portable fiber-based devices can provide a breakthrough platform for early diagnosis of cancer, infection and cardiovascular disorders with a potential to inhibit, if not prevent, metastasis, sepsis, and strokes or heart attack by well-timed personalized therapy.
传统的流式细胞术使用散射和荧光检测方法，多年来一直是生物学发现的基本工具。然而，从活体生物中侵入性地提取细胞可能导致细胞特性发生变化，并阻止对细胞在其自然环境中的长期研究。在这里，我们总结了新一代流式细胞术的最新进展，这些进展利用光声（PA）检测技术、多光谱高重复率激光、可调谐超锐（高达 0.8nm）彩虹等离子体纳米探针、正负 PA 对比、体内磁富集、飞行时间细胞速度测量、PA 光谱分析以及 PA、光热（PT）、荧光和拉曼方法的集成，用于在血液、淋巴、骨骼、大脑和植物脉管系统中进行非侵入性、无标记或靶向检测活体中的细胞。本文重点介绍了该工具的独特应用，包括对不同功能状态（如凋亡和坏死）的正常血细胞和罕见异常细胞（包括循环肿瘤细胞（CTC）、癌症干细胞、病原体、血栓、镰状细胞）进行超灵敏检测，以及对纳米颗粒、染料、微泡和药物纳米载体的药代动力学进行研究。使用该工具，我们发现触摸、活检或手术会增加 CTC 从原发性肿瘤中的释放，从而增加转移的风险。新型波动式流式细胞术为动态研究血液流变学提供了机会，包括不同医学条件下（如血液疾病、癌症或手术）红细胞聚集和血栓形成。光声诊断和 PT 纳米泡增强的多重治疗联合的治疗学为使用 PA 引导来控制治疗效率，消除 CTC、清除受感染的血液以及溶解血栓提供了机会。使用基于光纤的便携式设备进行体内流式细胞术，可为癌症、感染和心血管疾病的早期诊断提供一个突破性的平台，通过及时的个性化治疗，有可能抑制甚至预防转移、败血症和中风或心脏病发作。
