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自动化即时检测移动流式细胞术:将实验室带到样本身边。

Automated, Point-of-Care mobile flow cytometry: Bringing the laboratory to the sample.

作者信息

Jukema B N, Pelgrim T C, Spoelder M, Bongers C C W G, Hopman M T E, Smit K, Rijk M H, Venekamp R P, Vrisekoop N, Koenderman L

机构信息

Department of Respiratory Medicine, University Medical Center Utrecht, Utrecht University, Utrecht, Netherlands.

Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, Netherlands.

出版信息

Heliyon. 2024 Apr 3;10(8):e28883. doi: 10.1016/j.heliyon.2024.e28883. eCollection 2024 Apr 30.

Abstract

BACKGROUND

Innate effector cells are very responsive to infectious and inflammatory cues found in damaged and inflamed tissues. Their activation is a potential target to assess the state of the immune system. Unfortunately, these cells are very susceptible for ex-vivo activation, hampering accurate interpretation of flow cytometry data. Whether a brief window exists before ex-vivo activation starts to occur is currently unknown.

AIMS

  1. This study extensively investigated ex-vivo activation of innate effector cells over time. 2) We tested the feasibility of applying a mobile, automated, flow cytometry laboratory for out-of-hospital Point-of-Care analyses to minimize ex-vivo activation bias.

METHODS

  1. Ex-vivo neutrophil, eosinophil and monocyte activation in a blood collection tube over time and the reactivity to a formyl-peptide was investigated in a healthy cohort. 2) To facilitate fast, out-of-hospital analysis, application of the mobile flow cytometry was tested by placing an automated flow cytometer into a van. The stability of the setup was assessed by repetitively measuring laser alignment and fluorescence verification beads.

FINDINGS

  1. Immediately after venipuncture activation marker expression on neutrophils, eosinophils and monocyte subsets started to change in a time-dependent manner. 2) The mobile flow cytometry laboratory travelled over 3000 km, performing measurements at 19 locations with a median single-person-set-up time of 14 min. The laser alignment and fluorescence were stable during all experiments.

CONCLUSIONS

Accurate flow data of innate immune cells are only obtained when ex-vivo activation is kept to minimum. The use of a mobile, fast, automated, flow cytometry laboratory for out-of-hospital Point-of-Care analyses provides new investigational and diagnostic possibilities outside major hospital flow cytometry laboratories.

摘要

背景

固有效应细胞对受损和发炎组织中发现的感染性和炎症信号非常敏感。它们的激活是评估免疫系统状态的一个潜在靶点。不幸的是,这些细胞非常容易发生体外激活,这妨碍了对流式细胞术数据的准确解读。目前尚不清楚在体外激活开始发生之前是否存在一个短暂的窗口期。

目的

1)本研究广泛调查了固有效应细胞随时间的体外激活情况。2)我们测试了应用移动、自动化流式细胞术实验室进行院外即时检测分析以尽量减少体外激活偏差的可行性。

方法

1)在一个健康队列中,研究了采血管中中性粒细胞、嗜酸性粒细胞和单核细胞随时间的体外激活情况以及对甲酰肽的反应性。2)为便于快速的院外分析,通过将一台自动化流式细胞仪放置在一辆厢式货车中来测试移动流式细胞术的应用。通过重复测量激光校准和荧光验证微珠来评估该装置的稳定性。

研究结果

1)静脉穿刺后,中性粒细胞、嗜酸性粒细胞和单核细胞亚群上激活标志物的表达立即开始以时间依赖性方式发生变化。2)移动流式细胞术实验室行驶了超过3000公里,在19个地点进行测量,单人设置的中位时间为14分钟。在所有实验过程中,激光校准和荧光均保持稳定。

结论

只有将体外激活降至最低,才能获得固有免疫细胞的准确流式数据。使用移动、快速、自动化的流式细胞术实验室进行院外即时检测分析,为大型医院流式细胞术实验室之外的研究和诊断提供了新的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eab0/11019183/8d5b45db82ce/gr1.jpg

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