Brignole-Baudouin Françoise, Ott Ann-Christin, Warnet Jean-Michel, Baudouin Christophe
Laboratory of Toxicology, Faculty of Pharmaceutical and Biological Sciences, University of Paris-5 René Descartes, Paris, France.
Exp Eye Res. 2004 Mar;78(3):473-81. doi: 10.1016/j.exer.2003.08.005.
Flow cytometry (FCM) is a widely used technique, not only in immunology and hematology but also in many other domains and new applications have been developed in ophthalmology, for investigating ocular surface biology and pathophysiology. Apoptosis, proteins specific of cell lineage, expression of activation or immune-related markers, proinflammatory cytokine production, multi-drug resistance and oxidative stress are some examples of possible pathways that may be explored directly using ocular surface cells removed with the non-invasive technique of impression cytology, by means of flow cytometry. Conjunctival cells are thus collected by impression cytology on a filter, fixed in paraformaldehyde and dissociated by agitation before immunostaining and FCM processing. Important observations have been made concerning Sjögren's syndrome, ocular allergy and antiglaucoma eye drop toxicity. Studies performed on impression cytology with FCM techniques confirmed inflammatory and apoptotic patterns in a large number of patients with keratoconjunctivitis sicca or receiving long-term treatments with antiglaucoma drugs, even though they did not present any clinical signs of inflammation. FCM may now be used for large multicenter clinical trials and has already contributed to a better understanding of ocular surface pathophysiology, the monitoring of new immunomodulating drugs and knowledge concerning topical drugs, as regards both efficacy and safety.
流式细胞术（FCM）是一种广泛应用的技术，不仅用于免疫学和血液学，还用于许多其他领域，并且在眼科已经开发了新的应用，用于研究眼表生物学和病理生理学。细胞凋亡、细胞谱系特异性蛋白、激活或免疫相关标志物的表达、促炎细胞因子的产生、多药耐药性和氧化应激是一些可能的途径示例，这些途径可以通过印象细胞学的非侵入性技术采集眼表细胞，借助流式细胞术直接进行探索。结膜细胞通过印象细胞学方法收集在滤膜上，用多聚甲醛固定，在免疫染色和FCM处理前通过振荡解离。关于干燥综合征、眼部过敏和抗青光眼滴眼液毒性已经有了重要发现。使用FCM技术对印象细胞学进行的研究证实，大量患有干眼症或接受抗青光眼药物长期治疗的患者存在炎症和凋亡模式，即使他们没有任何炎症的临床体征。FCM现在可用于大型多中心临床试验，并且已经有助于更好地理解眼表病理生理学、监测新的免疫调节药物以及了解局部用药在疗效和安全性方面的知识。
