用于脑组织解剖和固定的小鼠经心脏灌注
Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation.
作者信息
Wu Jinyun, Cai Yuqi, Wu Xiaoyang, Ying Yue, Tai Yilin, He Miao
机构信息
Institutes of Brain Sciences, Fudan University, Shanghai 200032, China.
出版信息
Bio Protoc. 2021 Mar 5;11(5):e3988. doi: 10.21769/BioProtoc.3988.
Abstract
Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for performing this procedure in mouse.
摘要
先用生理盐水进行经心灌注，然后用4%多聚甲醛（PFA）灌注，这一方法被广泛用于清除血液并保存大脑，以进行免疫染色或杂交。PFA在溶液中分解为甲醛，甲醛可交联蛋白质和DNA分子以保存组织和细胞结构。在此，我们提供一份在小鼠中进行该操作的分步指南。
相似文献
1
Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation.用于脑组织解剖和固定的小鼠经心脏灌注
Bio Protoc. 2021 Mar 5;11(5):e3988. doi: 10.21769/BioProtoc.3988.
2
Aortic Cross-Clamping to Provide Differential Fixation by Perfusion.主动脉阻断以提供灌注差异固定。
Curr Protoc. 2021 Mar;1(3):e81. doi: 10.1002/cpz1.81.
3
The search for an optimal DNA, RNA, and protein detection by in situ hybridization, immunohistochemistry, and solution-based methods.原位杂交、免疫组织化学和基于溶液的方法检测 DNA、RNA 和蛋白质的最佳方法的研究。
Methods. 2010 Dec;52(4):281-6. doi: 10.1016/j.ymeth.2010.09.005. Epub 2010 Oct 1.
4
The Combination of Paraformaldehyde and Glutaraldehyde Is a Potential Fixative for Mitochondria.多聚甲醛和戊二醛的联合使用是一种潜在的线粒体固定剂。
Biomolecules. 2021 May 10;11(5):711. doi: 10.3390/biom11050711.
5
Effects of paraformaldehyde fixation on nicotinic acetylcholine receptor binding in adult and developing rat brain sections.多聚甲醛固定对成年及发育中大鼠脑切片烟碱型乙酰胆碱受体结合的影响。
J Neurosci Methods. 2006 Jun 15;153(2):312-7. doi: 10.1016/j.jneumeth.2005.11.008. Epub 2006 Jan 6.
6
The in situ morphology of microglia is highly sensitive to the mode of tissue fixation.小胶质细胞的原位形态对组织固定方式高度敏感。
J Chem Neuroanat. 2017 Dec;86:59-66. doi: 10.1016/j.jchemneu.2017.08.007. Epub 2017 Sep 1.
7
PFA is superior to glyoxal in preserving oocyte, embryo, and stem cell proteins evidenced by super-resolution microscopical surveys of epitopes.PFA 在保存卵母细胞、胚胎和干细胞蛋白方面优于乙二醛，这一点可以通过对表位的超分辨率显微镜研究得到证明。
J Assist Reprod Genet. 2020 Feb;37(2):369-384. doi: 10.1007/s10815-019-01666-9. Epub 2020 Jan 13.
8
Optimising Golgi-Cox staining for use with perfusion-fixed brain tissue validated in the zQ175 mouse model of Huntington's disease.优化用于灌注固定脑组织的高尔基-考克斯染色法，并在亨廷顿舞蹈病的zQ175小鼠模型中得到验证。
J Neurosci Methods. 2016 May 30;265:81-8. doi: 10.1016/j.jneumeth.2015.09.033. Epub 2015 Oct 13.
9
Acquisition of brains from the African elephant (Loxodonta africana): perfusion-fixation and dissection.获取非洲象（Loxodonta africana）的大脑：灌注固定与解剖。
J Neurosci Methods. 2009 Apr 30;179(1):16-21. doi: 10.1016/j.jneumeth.2009.01.001. Epub 2009 Jan 13.
10
Paraformaldehyde fixation of neutrophils for immunolabeling of granule antigens in cryoultrasections.用于冷冻超薄切片中颗粒抗原免疫标记的中性粒细胞多聚甲醛固定法。
J Histochem Cytochem. 1995 Oct;43(10):1019-25. doi: 10.1177/43.10.7560879.
引用本文的文献
1
Golgi-Cox Staining Protocol for Medium Spiny Neurons in the Striatum of Neonatal and Early Postnatal Mouse Brain Using Cryosections.使用冷冻切片对新生和出生后早期小鼠脑纹状体中型多棘神经元进行高尔基-考克斯染色的方案。
Int J Mol Sci. 2025 Aug 14;26(16):7870. doi: 10.3390/ijms26167870.
2
Development of a Novel Immunoprecipitation Method for Extracting Monoclonal Antibodies From Brain Tissue and Its Application to Assessing In Vivo Brain Penetration in Mouse via Liquid Chromatography-Mass Spectrometry.一种从脑组织中提取单克隆抗体的新型免疫沉淀方法的开发及其通过液相色谱-质谱法评估小鼠体内脑渗透的应用。
Biomed Chromatogr. 2025 Sep;39(9):e70196. doi: 10.1002/bmc.70196.
3
Glyco Ionic Liquids as Novel Nanoparticle Coatings to Enhance Triple-Negative Breast Cancer Drug Delivery.糖离子液体作为新型纳米颗粒涂层用于增强三阴性乳腺癌药物递送
Adv Healthc Mater. 2025 Jul 9:e2500592. doi: 10.1002/adhm.202500592.
4
Microscope-assisted, rodent whole heart perfusion fixation for histological characterization of myocardium remodeling.显微镜辅助的啮齿动物全心脏灌注固定用于心肌重塑的组织学特征分析。
MethodsX. 2025 Jun 3;14:103417. doi: 10.1016/j.mex.2025.103417. eCollection 2025 Jun.
5
Fluorescence-Guided Matrix-assisted Laser Desorption/Ionization with Laser-Induced Postionization Mass Spectrometry of Individual Rat Neural Cells.荧光引导的基质辅助激光解吸/电离结合激光诱导后电离质谱法对单个大鼠神经细胞的分析
J Vis Exp. 2025 May 23(219). doi: 10.3791/68376.
6
Differential retinal ganglion cell resilience to optic nerve injury across vertebrate species.脊椎动物物种中视网膜神经节细胞对视神经损伤的不同恢复能力。
Front Neurosci. 2025 May 20;19:1596464. doi: 10.3389/fnins.2025.1596464. eCollection 2025.
7
Protocol for mouse carotid artery perfusion for in situ brain tissue fixation and parallel unfixed tissue collection.用于原位脑组织固定和并行采集未固定组织的小鼠颈动脉灌注方案。
STAR Protoc. 2025 Mar 14;6(2):103699. doi: 10.1016/j.xpro.2025.103699.
8
C/EBP Homologous Protein Expression in Retinal Ganglion Cells Induces Neurodegeneration in Mice.视网膜神经节细胞中C/EBP同源蛋白的表达诱导小鼠神经退行性变。
Int J Mol Sci. 2025 Feb 21;26(5):1858. doi: 10.3390/ijms26051858.
9
Identification and Therapeutic Potential of Polymethoxylated Flavones in for Alzheimer's Disease: Targeting Neuroinflammation.聚甲氧基黄酮在阿尔茨海默病中的鉴定及其治疗潜力：针对神经炎症
Molecules. 2025 Feb 7;30(4):771. doi: 10.3390/molecules30040771.
10
Isolation of Live Myeloid and Epithelial Cell Populations from the Mouse Lung.从小鼠肺中分离活的髓样细胞和上皮细胞群体。
J Vis Exp. 2025 Jan 31(215). doi: 10.3791/67648.
本文引用的文献
1
Whole animal perfusion fixation for rodents.啮齿动物的全动物灌注固定
J Vis Exp. 2012 Jul 30(65):3564. doi: 10.3791/3564.
Zotero 插件