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新技术助力组织内蛋白成像
作者:小柯机器人 发布时间:2019/8/20 13:36:04

美国哈佛大学Peng Yin、Sinem K. Saka、Yu Wang等研究团队,合作研发了能够在组织中实现高度多通道和信号放大的蛋白质成像技术。相关论文于2019年8月19日在线发表在《自然—生物技术》上。

研究人员报道了通过交换反应进行信号放大的免疫染色(Immuno-SABER)技术,其通过DNA条形码化抗体和由引物交换反应(PER)产生的正交DNA多联体实现高度多重信号放大。SABRE提供独立的可编程信号放大,无需原位酶促反应,并具有内在的可扩展性,可在与荧光成像链的快速交换循环结合时快速放大和可视化大量目标。研究人员在不同的样品(培养的细胞、冷冻切片、福尔马林固定的石蜡包埋切片和整个组织)中实现了5到180倍的信号放大,并使用标准设备和工作流程对10种不同蛋白质进行同时信号扩增。研究人员还将SABER与显微镜相结合,以实现快速,多通道的超分辨组织成像。Immuno-SABER为蛋白质的多重和扩增成像提供了一个具有高灵敏度和高通量并且有效、方便使用的平台。

据了解,组织中蛋白质的空间定位受到多通道、敏感度和通量的限制。

附:英文原文

Title: Immuno-SABER enables highly multiplexed and amplified protein imaging in tissues

Author: Sinem K. Saka, Yu Wang, Jocelyn Y. Kishi, Allen Zhu, Yitian Zeng, Wenxin Xie, Koray Kirli, Clarence Yapp, Marcelo Cicconet, Brian J. Beliveau, Sylvain W. Lapan, Siyuan Yin, Millicent Lin, Edward S. Boyden, Pascal S. Kaeser, German Pihan, George M. Church, Peng Yin 

Issue&Volume: 19 August 2019

Abstract: Spatial mapping of proteins in tissues is hindered by limitations in multiplexing, sensitivity and throughput. Here we report immunostaining with signal amplification by exchange reaction (Immuno-SABER), which achieves highly multiplexed signal amplification via DNA-barcoded antibodies and orthogonal DNA concatemers generated by primer exchange reaction (PER). SABER offers independently programmable signal amplification without in situ enzymatic reactions, and intrinsic scalability to rapidly amplify and visualize a large number of targets when combined with fast exchange cycles of fluorescent imager strands. We demonstrate 5- to 180-fold signal amplification in diverse samples (cultured cells, cryosections, formalin-fixed paraffin-embedded sections and whole-mount tissues), as well as simultaneous signal amplification for ten different proteins using standard equipment and workflows. We also combined SABER with expansion microscopy to enable rapid, multiplexed super-resolution tissue imaging. Immuno-SABER presents an effective and accessible platform for multiplexed and amplified imaging of proteins with high sensitivity and throughput.

DOI: https://doi.org/10.1038/s41587-019-0207-y

Source: https://www.nature.com/articles/s41587-019-0207-y

期刊信息

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:31.864
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex