Department of Bioengineering, Imperial College London, London, UK.
J Microsc. 2024 Nov;296(2):154-161. doi: 10.1111/jmi.13244. Epub 2023 Nov 15.
In 2015, we reported primed conversion, a novel way to convert green-to-red photoconvertible fluorescent proteins, which emerges as a powerful tool for precision optical imaging. Primed conversion uses the intercept of blue and red-to-far-red light instead of traditional violet or near-UV light illumination which offers a series of advantages. Here, we review the fundamental principles and applications of primed conversion with a focus on its use in single-cell labelling and lineage tracing. We provide a historical perspective of lineage tracing techniques, thereby covering basic principles of fluorescence, photoconvertible fluorescent proteins, and eventually primed conversion. We then present the molecular requirements for primed conversion to take place and showcase how it can be used for dual-colour high-fidelity lineage tracing. Further, we discuss potential future developments of the primed conversion imaging toolkit that can benefit the study of both development and disease progression.
2015 年，我们报道了引物转换，这是一种将绿色到红色光转化的荧光蛋白进行转换的新方法，它成为精确光学成像的强大工具。引物转换使用蓝红远红光的交叉取代传统的紫或近紫外光照射，具有一系列优势。在这里，我们回顾了引物转换的基本原理和应用，重点介绍了它在单细胞标记和谱系追踪中的应用。我们提供了谱系追踪技术的历史视角，从而涵盖了荧光、光转化荧光蛋白的基本原理，最终介绍了引物转换。然后，我们介绍了引物转换发生的分子要求，并展示了如何将其用于双色高保真谱系追踪。此外，我们还讨论了引物转换成像工具包的潜在未来发展，这将有助于研究发育和疾病进展。
