单细胞测序

|   技术原理

10x Genomics平台核心基于“微流控油包水（GEMs）+条形码（Barcode）”技术体系，将待分析细胞悬液、含Barcode的凝胶珠（Gel Beads）、酶试剂一同注入微流控芯片，形成油包水的微体系（GEMs）；随后，凝胶珠溶解释放Barcode，细胞裂解后释放的RNA与Barcode结合，通过逆转录生成带UMI标记的cDNA；随后收集所有微滴的cDNA产物，进行 PCR 扩增、片段化、接头连接等标准化文库构建步骤；其中barcode用于区分细胞，UMI用于区分mRNA分子。

|   检测流程

从单细胞悬液制备到最终数据获得，单细胞RNA测序实验流程主要包括：单细胞悬液制备和质检、生成GEMs、反转录成cDNA、cDNA回收扩增、cDNA质检、文库构建以及上机测序。实验流程中的每一环节都会直接影响数据质量，从而影响后续数据分析的结果。因此，为保证测序数据准确可靠，华盈生物在数据生产的所有环节都严格把关，从而确保高质量数据的产出。建库测序的流程图如下所示。

|   技术优势

1.高通量：一张芯片具有8个独立的通道，可供8个样本同时上机，每个通道最高可捕获10000个细胞。

2.高捕获效率与灵敏度单细胞捕获效率高达 65%，多细胞比例低至约0.9%每1000个细胞。

3.多个地区均有单细胞驻地实验室，大大解决新鲜组织样本解离的时效性问题

4.完成大部分样本类型的单细胞制备，经验丰富

|   数据分析

|   相关文献

1. Chang J, Lu J, Liu Q, et al. Single-cell multi-stage spatial evolutional map of esophageal carcinogenesis. Cancer Cell. 2025;43(3):380-397.e7.

2. Li Q, Xiao Y, Han L, et al. Microbiome dysbiosis, neutrophil recruitment and mesenchymal transition of mesothelial cells promotes peritoneal metastasis of colorectal cancer. Nat Cancer. 2025;6(3):493-510.

3. Xing X, Zhong J, Biermann J, et al. Pan-cancer human brain metastases atlas at single-cell resolution. Cancer Cell. 2025;43(7):1242-1260.e9.

84 Yan Y, Sun D, Hu J, et al. Multi-omic profiling highlights factors associated with resistance to immuno-chemotherapy in non-small-cell lung cancer. Nat Genet. 2025;57(1):126-139.

5. Qiu X, Zhou T, Li S, et al. Spatial single-cell protein landscape reveals vimentinhigh macrophages as immune-suppressive in the microenvironment of hepatocellular carcinoma. Nat Cancer. 2024;5(10):1557-1578.

6. Liu W, You W, Lan Z, et al. An immune cell map of human lung denocarcinoma development reveals an anti-tumoral role of the Tfh-dependent tertiary lymphoid structure. Cell Rep Med. 2024;5(3):101448.

7. Jiang YZ, Ma D, Jin X, et al. Integrated multiomic profiling of breast cancer in the Chinese population reveals patient stratification and therapeutic vulnerabilities. Nat Cancer. 2024;5(4):673-690.

8. Zhang Y, Liu G, Zeng Q, et al. CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis. Cancer Cell. 2024;42(8):1370-1385.e9.

9. Jiang L, Qin J, Dai Y, et al. Prospective observational study on biomarkers of response in pancreatic ductal adenocarcinoma. Nat Med. 2024;30(3):749-761.

10. Mei Y, Wang X, Zhang J, et al. Siglec-9 acts as an immune-checkpoint molecule on macrophages in glioblastoma, restricting T-cell priming and immunotherapy response. Nat Cancer. 2023;4(9):1273-1291.