Intracellular delivery of advanced therapeutics, including biologicals and supramolecular agents, is complex because of the natural biological barriers that have evolved to protect the cell. Efficient delivery of therapeutic nucleic acids, proteins, peptides and nanoparticles is crucial for clinical adoption of emerging technologies that can benefit disease treatment through gene and cell therapy. Nanoneedles are arrays of vertical high-aspect-ratio nanostructures that can precisely manipulate complex processes at the cell interface, enabling effective intracellular delivery. This emerging technology has already enabled the development of efficient and non-destructive routes for direct access to intracellular environments and delivery of cell-impermeant payloads. However, successful implementation of this technology requires knowledge of several scientific fields, making it complex to access and adopt by researchers who are not directly involved in developing nanoneedle platforms. This presents an obstacle to the widespread adoption of nanoneedle technologies for drug delivery. This tutorial aims to equip researchers with the knowledge required to develop a nanoinjection workflow. It discusses the selection of nanoneedle devices, approaches for cargo loading and strategies for interfacing to biological systems and summarises an array of bioassays that can be used to evaluate the efficacy of intracellular delivery.

高级疗法（包括生物制剂和超分子制剂）的细胞内递送非常复杂，因为已经进化出保护细胞的天然生物屏障。有效递送治疗性核酸、蛋白质、肽和纳米颗粒对于临床采用新兴技术至关重要，这些技术可以通过基因和细胞疗法有益于疾病治疗。纳米针是垂直的高纵横比纳米结构阵列，可以精确地操纵细胞界面的复杂过程，从而实现有效的细胞内递送。这种新兴技术已经能够开发出有效且无损的途径，用于直接进入细胞内环境和传递细胞不可渗透的有效载荷。然而，这项技术的成功实施需要多个科学领域的知识，这使得不直接参与开发纳米针平台的研究人员难以访问和采用。这对广泛采用纳米针技术进行药物输送构成了障碍。本教程旨在为研究人员提供开发纳米注射工作流程所需的知识。它讨论了纳米针装置的选择、货物装载的方法和与生物系统接口的策略，并总结了一系列可用于评估细胞内递送功效的生物测定。这对广泛采用纳米针技术进行药物输送构成了障碍。本教程旨在为研究人员提供开发纳米注射工作流程所需的知识。它讨论了纳米针装置的选择、货物装载的方法和与生物系统接口的策略，并总结了一系列可用于评估细胞内递送功效的生物测定。这对广泛采用纳米针技术进行药物输送构成了障碍。本教程旨在为研究人员提供开发纳米注射工作流程所需的知识。它讨论了纳米针装置的选择、货物装载的方法和与生物系统接口的策略，并总结了一系列可用于评估细胞内递送功效的生物测定。