众所周知，实体瘤的生长和转移是由肿瘤微环境（TME）促进的，肿瘤微环境由高度多样化的细胞类型集合组成，这些细胞类型之间广泛相互作用和交流。其中许多相互作用涉及 TME 内的免疫细胞群，称为肿瘤免疫微环境 (TIME)。这些非细胞自主相互作用对细胞行为产生重大影响，并有助于将免疫细胞和基质细胞重编程为多种促肿瘤表型。对其中一些相互作用的研究，例如诱导 CD8 T 细胞耗竭的 PD-1/PD-L1 轴，已经取得了突破性的治疗进展。然而，许多常见的 TME 分析要么不保留评估细胞间相互作用所需的空间数据，要么询问很少（<10）的标记，从而限制了细胞表型分析的能力。最近开发的数字病理学技术与复杂的生物图像分析程序一起，现在可以对临床标本 TME 内的各种免疫和基质细胞标记物进行高分辨率、高度多重分析。在本文中，我们回顾了 TME 中促进肿瘤的非细胞自主相互作用及其对肿瘤行为的影响。我们还调查了常用的图像分析程序和高度复用的空间成像技术，并讨论了它们的相对优势和局限性。 TME 的空间组织因患者而异，因此在未来的研究中利用这些技术来进一步描述非细胞自主相互作用如何影响肿瘤行为可能会为癌症治疗的个性化提供信息。版权所有 © 2023 Cohn、Forder、Marshall，武契奇、斯图尔特、努尔丁、洛克伍德、麦考利、吉约和林。

The growth and metastasis of solid tumours is known to be facilitated by the tumour microenvironment (TME), which is composed of a highly diverse collection of cell types that interact and communicate with one another extensively. Many of these interactions involve the immune cell population within the TME, referred to as the tumour immune microenvironment (TIME). These non-cell autonomous interactions exert substantial influence over cell behaviour and contribute to the reprogramming of immune and stromal cells into numerous pro-tumourigenic phenotypes. The study of some of these interactions, such as the PD-1/PD-L1 axis that induces CD8+ T cell exhaustion, has led to the development of breakthrough therapeutic advances. Yet many common analyses of the TME either do not retain the spatial data necessary to assess cell-cell interactions, or interrogate few (<10) markers, limiting the capacity for cell phenotyping. Recently developed digital pathology technologies, together with sophisticated bioimage analysis programs, now enable the high-resolution, highly-multiplexed analysis of diverse immune and stromal cell markers within the TME of clinical specimens. In this article, we review the tumour-promoting non-cell autonomous interactions in the TME and their impact on tumour behaviour. We additionally survey commonly used image analysis programs and highly-multiplexed spatial imaging technologies, and we discuss their relative advantages and limitations. The spatial organization of the TME varies enormously between patients, and so leveraging these technologies in future studies to further characterize how non-cell autonomous interactions impact tumour behaviour may inform the personalization of cancer treatment.​.Copyright © 2023 Cohn, Forder, Marshall, Vucic, Stewart, Noureddine, Lockwood, MacAulay, Guillaud and Lam.