Platelet morphology, ultrastructure and function changes in acute ischemic stroke patients based on structured illumination microscopy
Bingxin Yang,
Xifeng Wang,
Xiaoyu Hu,
Yao Xiao,
Xueyu Xu,
Xiaomei Yu,
Min Wang,
Honglian Luo,
Jun Li,
Wei Shen,
Yan Ma
Affiliations
Bingxin Yang
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China
Xifeng Wang
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Xiaoyu Hu
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Yao Xiao
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Xueyu Xu
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Xiaomei Yu
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Min Wang
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Honglian Luo
Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Jun Li
Corresponding author.; Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Wei Shen
Corresponding author.; Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Yan Ma
Corresponding author.; Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China; Wuhan Forth Hospital, Wuhan, Hubei, 430030, China; Wuhan Blood Center-Huazhong University of Science and Technology United Hematology Optical Imaging Center, Hubei Institute of Blood Transfusion, Wuhan Blood Center, Wuhan, Hubei, 430030, China
Acute ischemic stroke (AIS) is the second leading cause of death worldwide. This study aims at assessing platelet morphology, ultrastructure and function changes of platelets in acute ischemic stroke (AIS) patients by the technique of Structured Illumination Microscopy (SIM). This assay collected platelet-rich plasma (PRP) from 11 AIS patients and 12 healthy controls. Each PRP sample was divided into 7 groups:1) rest group; 2) Thrombin-treated 5 min group; 3) Thrombin plus 2MeSAMP-treated 5 min group; 4) Thrombin plus Aspirin-treated 5 min group; 5) Thrombin-treated 1 h group; 6) Thrombin plus 2MeSAMP-treated 1 h group; 7) Thrombin plus Aspirin-treated 1 h group. SIM was applied to observe dense granules and α-granules morphology changes of platelet in AIS patients. FIJI was used to quantify the image data. We finally observed 1448 images of platelets within the 7 groups. In rest group, 7162 platelets were calculated platelet diameter, CD63 dots, average CD63-positive dots area, CD63-positive area per platelet, CD63-positive area Fov, VWF dots, average VWF-positive dots area, VWF-positive area per platelet and VWF-positive area Fov. ELISA was used to detect release of platelet factor 4 (PF4) of α-granules. The results showed that AIS patients had lower number and smaller area of platelet granules. Platelet α-granules of AIS patients concentrated to parenchymal-like fluorescent blocks in Thrombin-treated 1 h group. Antiplatelet drug treatment could reverse the concentration of platelets α-granules, and 2MeSAMP was more powerful than Aspirin in vitro. This study complemented detail information of platelet ultrastructure of AIS patients, provided a new perspective on the pathogenesis of AIS and the mechanism of antiplatelet drugs based on SIM and provided a reference for future related studies. SIM-based analysis of platelet ultrastructure may be useful for detecting antiplatelet drugs and AIS in the future.
