Immunofluorescence Protocol for Characterization of Platelet and Leukocyte Binding in Extracorporeal Membrane Oxygenation (ECMO) Circuits

被引:0
作者
Cai, Tengyi [1 ,2 ]
Burton, Matthew [3 ]
Mccafferty, Conor [1 ,2 ]
van den Helm, Suelyn [1 ,2 ]
Letunica, Natasha [1 ]
Attard, Chantal [1 ,2 ]
Horton, Stephen [2 ,4 ]
Bottrell, Steve [4 ]
Schultz, Bradley [4 ]
Maclaren, Graeme [5 ]
Chiletti, Roberto [6 ,7 ]
Best, Derek [6 ,7 ]
Johansen, Amy [6 ,7 ]
Newall, Fiona [1 ,2 ,8 ]
Butt, Warwick [2 ,6 ,7 ]
d'Udekem, Yves [9 ]
Monagle, Paul [1 ,2 ,8 ,10 ]
Ignjatovic, Vera [1 ,2 ,4 ,11 ,12 ]
机构
[1] Murdoch Childrens Res Inst, Haematol Res, Parkville, WA, Australia
[2] UNIV MELBOURNE, DEPT PAEDIAT, PARKVILLE, Australia
[3] Murdoch Childrens Res Inst, Flow Cytometry & Imaging, Parkville, Australia
[4] Royal Childrens Hosp, Cardiac Surg, Parkville, Australia
[5] Natl Univ Hlth Syst, Cardiothorac Intens Care Unit, Singapore, Singapore
[6] Royal Childrens Hosp, Dept Neonatol, Parkville, Australia
[7] Murdoch Childrens Res Inst, Hematol Res Grp, Parkville, Vic, Australia
[8] Royal Childrens Hosp, Clin Haematol, Parkville, Australia
[9] Childrens Natl Hosp, Childrens Natl Heart Inst, Washington, DC USA
[10] Sydney Childrens Hosp, Kids Canc Ctr, Randwick, Australia
[11] Johns Hopkins All Childrens Inst Clin & Translat R, 600 5th St South,Suite 3200, St Petersburg, FL 33701 USA
[12] Johns Hopkins Univ, Sch Med, Dept Pediat, Baltimore, MD USA
基金
英国医学研究理事会;
关键词
ECMO; immunofluorescence; critical care; procedures; pediatric; THROMBOSIS;
D O I
10.1097/MAT.0000000000002259
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
The continuous contact between blood and the foreign surface of the extracorporeal membrane oxygenation (ECMO) circuit contributes to hemostatic, inflammatory, and other physiological disturbances observed during ECMO. Although previous studies have extensively investigated blood samples from patients on ECMO, cell adsorption to the ECMO circuit as an additional factor that could potentially influence clinical outcomes, has largely been overlooked. Here we provide a detailed immunofluorescence (IF) protocol designed to characterize cellular binding on ECMO circuits collected from patients. Extracorporeal membrane oxygenation circuits were collected from three pediatric patients and an albumin primed-only ECMO circuit was used as control. Circuit samples from five different sites within each ECMO circuit were collected and processed for the IF protocol. CD14 and CD42a antibodies were used to identify platelets and leukocytes bound to each ECMO circuit sample and images captured using inverted fluorescence microscopy. The protocol enables the comprehensive characterization of platelet and leukocyte binding to ECMO circuits collected from patients, which could in turn extend our knowledge of the characteristics of circuit binding and may provide guidance for improved ECMO circuit design.
引用
收藏
页码:e1 / e4
页数:4
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