The microinjection of fluorescent probes into live cells is an essential component in the toolbox of modern cell biology. Microinjection techniques include the penetration of the plasma membrane and, if present, the cell wall with micropipettes, and the application of pressure or electrical currents to drive the micropipette contents into the cell. These procedures interfere with cellular functions and therefore may induce artifacts. We designed the diffusive injection micropipette (DIMP) that avoids most of the possible artifacts due to the drastically reduced volume of its fluid contents and the utilization of diffusion for cargo delivery into the target cell. DIMPs were successfully tested in plant, fungal, and animal cells. Using the continuity of cytoplasmic dynamics over ten minutes after impalement of Nicotiana trichome cells as a criterion for non-invasiveness, we found DIMPs significantly less disruptive than conventional pressure microinjection. The design of DIMPs abolishes major sources of artifacts that cannot be avoided by other microinjection techniques. Moreover, DIMPs are inexpensive, easy to produce, and can be applied without specific equipment other than a micromanipulator. With these features, DIMPs may become the tool of choice for studies that require the least invasive delivery possible of materials into live cells.

中文翻译：

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扩散注射微量移液器 (DIMP)

将荧光探针显微注射到活细胞中是现代细胞生物学工具箱中的重要组成部分。显微注射技术包括用微量移液管穿透质膜和（如果存在）细胞壁，以及施加压力或电流以将微量移液器内容物驱动到细胞中。这些程序会干扰细胞功能，因此可能会产生伪影。我们设计了扩散注射微量移液器 (DIMP)，它避免了大多数可能的伪影，因为其流体内容物的体积急剧减少，并且利用扩散将货物输送到目标细胞中。DIMP 已在植物、真菌和动物细胞中成功测试。使用刺入烟草毛状体细胞后 10 分钟内细胞质动力学的连续性作为非侵入性的标准，我们发现 DIMP 的破坏性明显低于传统压力显微注射。DIMP 的设计消除了其他显微注射技术无法避免的主要伪影来源。此外，DIMP 价格低廉，易于生产，并且可以在没有显微操作器以外的特定设备的情况下应用。凭借这些特性，DIMP 可能成为研究的首选工具，这些研究需要尽可能少地将材料输送到活细胞中。DIMP 价格低廉、易于生产，并且可以在没有显微操作器以外的特定设备的情况下进行应用。凭借这些特性，DIMP 可能成为研究的首选工具，这些研究需要尽可能少地将材料输送到活细胞中。DIMP 价格低廉、易于生产，并且可以在没有显微操作器以外的特定设备的情况下进行应用。凭借这些特性，DIMP 可能成为研究的首选工具，这些研究需要尽可能少地将材料输送到活细胞中。