Palmer William M, Martin Antony P, Flynn Jamie R, Reed Stephanie L, White Rosemary G, Furbank Robert T, Grof Christopher P L
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia.
School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308, Australia.
Sci Rep. 2015 Sep 2;5:13492. doi: 10.1038/srep13492.
Here we report the adaptation of the CLARITY technique to plant tissues with addition of enzymatic degradation to improve optical clearing and facilitate antibody probe penetration. Plant-Enzyme-Assisted (PEA)-CLARITY, has allowed deep optical visualisation of stains, expressed fluorescent proteins and IgG-antibodies in Tobacco and Arabidopsis leaves. Enzyme treatment enabled penetration of antibodies into whole tissues without the need for any sectioning of the material, thus facilitating protein localisation of intact tissue in 3D whilst retaining cellular structure.
在此,我们报告了将CLARITY技术应用于植物组织,并添加酶解作用以改善光学透明效果并促进抗体探针渗透。植物酶辅助(PEA)-CLARITY技术实现了对烟草和拟南芥叶片中染色剂、表达的荧光蛋白和IgG抗体的深度光学可视化。酶处理使得抗体能够穿透整个组织,而无需对材料进行任何切片,从而便于在三维空间中对完整组织进行蛋白质定位,同时保留细胞结构。