美国哈佛大学Peng Yin团队利用DNA编码热通道进行无流体、快速连续多重成像。这一研究成果于2023年12月27日在线发表在国际学术期刊《自然—方法学》上。

研究人员报告的thermal-plex省去了复杂而缓慢的缓冲液交换步骤，提供了无流体的快速顺序成像。thermal-plex使用简单的DNA探针，当且仅当探针在指定温度（热通道）下瞬时暴露于加热脉冲而被激活时，探针才会依次发出荧光。通道切换速度快（<30 秒），可通过市场上可买到的价格合理的镜下加热装置实现。研究人员在固定细胞和视网膜组织中展示了15种RNA成像（5个热通道×3个荧光通道），耗时不到4分钟，无需使用缓冲液交换或流体技术。thermal-plex为高效的连续多路复用成像引入了一种新的标记方法。

据了解，在标准装置上，多重荧光成像通常仅限于三至五个复合物。基于迭代标记和成像的顺序成像方法可实现更高的实际多重性，但通常需要复杂的流体设置和几轮缓慢的缓冲液交换（每个交换步骤需要几十分钟到一小时）。

附：英文原文

Title: Thermal-plex: fluidic-free, rapid sequential multiplexed imaging with DNA-encoded thermal channels

Author: Hong, Fan, Kishi, Jocelyn Y., Delgado, Ryan N., Jeong, Jiyoun, Saka, Sinem K., Su, Hanquan, Cepko, Constance L., Yin, Peng

Issue&Volume: 2023-12-27

Abstract: Multiplexed fluorescence imaging is typically limited to three- to five-plex on standard setups. Sequential imaging methods based on iterative labeling and imaging enable practical higher multiplexing, but generally require a complex fluidic setup with several rounds of slow buffer exchange (tens of minutes to an hour for each exchange step). We report the thermal-plex method, which removes complex and slow buffer exchange steps and provides fluidic-free, rapid sequential imaging. Thermal-plex uses simple DNA probes that are engineered to fluoresce sequentially when, and only when, activated with transient exposure to heating spikes at designated temperatures (thermal channels). Channel switching is fast (<30s) and is achieved with a commercially available and affordable on-scope heating device. We demonstrate 15-plex RNA imaging (five thermal×three fluorescence channels) in fixed cells and retina tissues in less than 4min, without using buffer exchange or fluidics. Thermal-plex introduces a new labeling method for efficient sequential multiplexed imaging.

DOI: 10.1038/s41592-023-02115-3

Source: https://www.nature.com/articles/s41592-023-02115-3