Blocking autofluorescence in brain tissues affected by ischemic stroke, hemorrhagic stroke, or traumatic brain injury

Frontiers in Immunology

Immunology

Changsheng Li¹  Xiaoyu Niu¹  Chunhua Song²  Junmin Wang³  Qinfeng Peng³  Xiuhua Ren³  Weidong Zang³  Xuemei Chen³  Chao Jiang⁴  Xiaochong Fan⁵  Jian Wang⁶  Shaoshuai Wang⁶  Marietta Zille⁷ 

[1] Department of Anesthesiology and Perioperative Medicine, Affiliated Cancer Hospital of Zhengzhou University, Henan, China;Department of Epidemiology and Statistics, College of Public Health, Zhengzhou University, Zhengzhou, Henan, China;Department of Human Anatomy, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China;Department of Neurology, The Fifth Affiliated Hospital of Zhengzhou University, Henan, China;Department of Pain Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China;Department of Pain Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China;Department of Human Anatomy, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China;Department of Pharmaceutical Sciences, Division of Pharmacology and Toxicology, University of Vienna, Vienna, Austria;

关键词: autofluorescence;    Sudan black B;    intracerebral hemorrhage;    traumatic brain injury;    ICH;    cerebral ischemia;   

DOI  :  10.3389/fimmu.2023.1168292

 received in 2023-02-17, accepted in 2023-05-04,  发布年份 2023

来源: Frontiers

【 摘 要 】

Autofluorescence is frequently observed in animal tissues, interfering with an experimental analysis and leading to inaccurate results. Sudan black B (SBB) is a staining dye widely used in histological studies to eliminate autofluorescence. In this study, our objective was to characterize brain tissue autofluorescence present in three models of acute brain injury, including collagenase-induced intracerebral hemorrhage (ICH), traumatic brain injury (TBI), and middle cerebral artery occlusion, and to establish a simple method to block autofluorescence effectively. Using fluorescence microscopy, we examined autofluorescence in brain sections affected by ICH and TBI. In addition, we optimized a protocol to block autofluorescence with SBB pretreatment and evaluated the reduction in fluorescence intensity. Compared to untreated, pretreatment with SBB reduced brain tissue autofluorescence in the ICH model by 73.68% (FITC), 76.05% (Tx Red), and 71.88% (DAPI), respectively. In the TBI model, the ratio of pretreatment to untreated decreased by 56.85% (FITC), 44.28% (Tx Red), and 46.36% (DAPI), respectively. Furthermore, we tested the applicability of the protocol using immunofluorescence staining or Cyanine-5.5 labeling in the three models. SBB treatment is highly effective and can be applied to immunofluorescence and fluorescence label imaging techniques. SBB pretreatment effectively reduced background fluorescence but did not significantly reduce the specific fluorescence signal and greatly improved the signal-to-noise ratio of fluorescence imaging. In conclusion, the optimized SBB pretreatment protocol blocks brain section autofluorescence of the three acute brain injury models.

【 授权许可】

Unknown   
Copyright © 2023 Wang, Ren, Wang, Peng, Niu, Song, Li, Jiang, Zang, Zille, Fan, Chen and Wang