肿瘤相关的第三类淋巴结构（TLSs）是肿瘤组织内的异位淋巴结形成，主要由B细胞和T细胞群体形成有机聚集体。TLSs在肿瘤中的存在与患者对免疫疗法方案的响应以及肿瘤预后的改善密切相关。研究人员积极探索TLSs的原因是它们在临床应用方面的远景广阔。多项研究试图解析TLSs的形成机制、结构组成、诱导产生、预测标记和临床应用。同时，定性和定量描述的科学方法对于TLS研究至关重要。在检测方面，溴化镉和噪声免疫组化、多重免疫组织化学、多重免疫荧光和12种趋化因子基因签名已被认可为首选方法。然而，目前还没有标准的方法来定量分析TLSs，例如绝对TLS计数、TLS成分细胞的分析、结构特征、TLS空间位置、密度和成熟度。本研究回顾了TLS检测和定量的最新研究进展，提出了TLS评估的新方向，并解决了TLSs在临床定量应用中的问题。

Tumor-associated tertiary lymphoid structures (TLSs) are ectopic lymphoid formations within tumor tissue, with mainly B and T cell populations forming the organic aggregates. The presence of TLSs in tumors has been strongly associated with patient responsiveness to immunotherapy regimens and improving tumor prognosis. Researchers have been motivated to actively explore TLSs due to their bright clinical application prospects. Various studies have attempted to decipher TLSs regarding their formation mechanism, structural composition, induction generation, predictive markers, and clinical utilization. Meanwhile, the scientific approaches to qualitative and quantitative descriptions are crucial for TLS studies. In terms of detection, hematoxylin and eosin (H&E), multiplex immunohistochemistry (mIHC), multiplex immunofluorescence (mIF), and 12-chemokine gene signature have been the top approved methods. However, no standard methods exist for the quantitative analysis of TLSs, such as absolute TLS count, analysis of TLS constituent cells, structural features, TLS spatial location, density, and maturity. This study reviews the latest research progress on TLS detection and quantification, proposes new directions for TLS assessment, and addresses issues for the quantitative application of TLSs in the clinic.