Thermal-plex: fluidic-free, rapid sequential multiplexed imaging with DNA-encoded thermal channels.
Thermal-plex: fluidic-free, rapid sequential multiplexed imaging with DNA-encoded thermal channels.
复制标题
Thermal-plex：采用 DNA 编码热通道的无流体、快速顺序多重成像。
DOI：
10.1038/s41592-023-02115-3
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发表时间：
2024
期刊：
影响因子：
48
通讯作者：
Yin,Peng
中科院分区：
文献类型：
--
作者：
Hong,Fan;Kishi,JocelynY;Delgado,RyanN;Jeong,Jiyoun;Saka,SinemK;Su,Hanquan;Cepko,ConstanceL;Yin,Peng
多路荧光成像通常限于三至五路标准设置。基于迭代标记和成像的顺序成像方法实现了实际的更高的多路复用，但通常需要复杂的流体设置，并进行几轮缓慢的缓冲交换（每个交换步骤需要数十分钟到一小时）。我们报道了热复形方法，它消除了复杂和缓慢的缓冲交换步骤，并提供了无流体的快速顺序成像。thermal -plex使用简单的DNA探针，当且仅当在指定温度（热通道）下短暂暴露于加热峰值时激活，该探针被设计成顺序荧光。通道切换速度快（<30秒），可通过市售且价格合理的范围内加热装置实现。我们在固定细胞和视网膜组织中展示了15-plex RNA成像（5个热x 3个荧光通道），在不到4分钟的时间内，不使用缓冲交换或流体。Thermal-plex介绍了一种新的标记方法，用于高效的顺序多路成像。
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 (<30 s) 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 4 min, without using buffer exchange or fluidics. Thermal-plex introduces a new labeling method for efficient sequential multiplexed imaging.