样品分析
概述
准备样品（微孔板孔）从板上取下液体
加入100μL/孔的iFluor 594-鬼笔环肽溶液
在室温下染色细胞15到60分钟洗涤细胞
在显微镜下观察样品（Ex / Em = 583/603 nm）
注意：打开前将所有组件加热至室温
操作步骤
1.准备1X iFluor 594-鬼笔环肽工作溶液：
将10μL的iFluor 594-鬼笔环肽（组分A）加入10mL标记缓冲液（组分B）中。
注意1：未使用的1X iFluor 594-鬼笔环肽原液应等分并保存在-20 ℃，避光。
注意2：不同的细胞类型可能染色不同。 应相应地制备iFluor 594-鬼笔环肽工作溶液的浓度。
2.染色细胞：
2.1执行甲醛固定。 在室温下用3.0-4.0％甲醛的PBS孵育细胞10-30分钟。
注意：避免使用任何含甲醇的固定剂，因为甲醇会在固定过程中破坏肌动蛋白。 优选的固定剂是不含甲醇的甲醛。
2.2用PBS冲洗固定的细胞2-3次。
2.3可选：在PBS中加入0.1％Triton X-100固定细胞（步骤2.2）3至5分钟，以增加渗透性。 用PBS冲洗细胞2-3次。
2.4将100μL/孔（96孔板）的iFluor 594-鬼笔环肽工作溶液（来自步骤1）加入固定的细胞（来自步骤2.2或2.3），并在室温下将细胞染色15至60分钟。
2.5用PBS轻轻冲洗细胞2至3次以去除多余的染料，然后进行板密封和成像。
试剂应用文献
Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization
Authors: Yutong He, Genlan Ye, Chen Song, Chuangkun Li, Weirong Xiong, Lei Yu, Xiaozhong Qiu, Leyu Wang
Journal: Theranostics. 2018; 8(18): 5159–5177.
参考文献
Porous Li-containing biphasic calcium phosphate scaffolds fabricated by three-dimensional plotting for bone repair
Authors: Xiaoheng Guo, Huichang Gao, Xiao Liu, Jingjing Diao, Xuetao Shi, Naru Zhao, Yingjun Wang
Journal: RSC Advances (2017): 34508–34516
Preparation, characterization and in vitro cell performance of anti-washout calcium phosphate cement modified by sodium polyacrylate
Authors: Xingmei Li, Fupo He, Jiandong Ye
Journal: RSC Advances (2017): 32842–32849
miR-29b-Loaded Gold Nanoparticles Targeting to the Endoplasmic Reticulum for Synergistic Promotion of Osteogenic Differentiation
Authors: Ting Pan, Wenjing Song, Huichang Gao, Tianjie Li, Xiaodong Cao, Shizhen Zhong, Yingjun Wang
Journal: ACS Applied Materials & Interfaces (2016): 19217–19227
Osteogenic and tenogenic induction of hBMSCs by integrated nanofibrous scaffold with chemical and structural mimic to bone-ligament connection
Authors: Zifeng Lin, Xiujuan Zhao, Si Chen, Chang Du
Journal: Journal of Materials Chemistry B (2016)
The stimulation of the differentiation of pheochromocytoma (PC12-L) cells into neuron-like cells by electrically conductive nanofibre mesh
Authors: Huishang Yang, Guanglin Zhu, Yicheng Huang, Xuetao Shi, Yingjun Wang
Journal: Applied Materials Today (2016): 215–222
Distinct mechanical behavior of HEK293 cells in adherent and suspended states
Authors: Seyed Mohammad Ali Haghparast, Takanori Kihara, Jun Miyake
Journal: PeerJ (2015): e1131
The preparation and characterization of polycaprolactone/graphene oxide biocomposite nanofiber scaffolds and their application for directing cell behaviors
Authors: Juqing Song, Huichang Gao, Guanglin Zhu, Xiaodong Cao, Xuetao Shi, Yingjun Wang
Journal: Carbon (2015): 1039–1050
Actin-based biomechanical features of suspended normal and cancer cells
Authors: Seyed Mohammad Ali Haghparast, Takanori Kihara, Yuji Shimizu, Shunsuke Yuba, Jun Miyake
Journal: Journal of bioscience and bioengineering (2013): 380–385
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