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Abstract：
UNASSIGNED: Spatial transcriptome (ST) provides molecular profiles of tumor cells at the spatial level, which brings new progress to the research of tumors and the tumor microenvironment. This study summarizes the experiences and lessons learned in the spatial section preparation of two different pathological types of nose and skull base tumors at our institution, with the aim of offering guidelines to researchers to avoid wasting precious samples and provide a basis for the application of ST in clinical practice.
UNASSIGNED: Frozen tissue blocks from patients with squamous cell carcinoma and adenocarcinoma of the nose and skull base diagnosed at our institution were prepared. The effects of different procedures and pathological tissue types on slide quality were explored and evaluated using RNA integrity number (RIN) and HE scores as criteria. The effects of different RIN values on ST sequencing data were explored.
UNASSIGNED: A total of 43 samples were obtained from 26 patients, including 22 with squamous carcinomas and 21 with adenocarcinomas. Thirteen samples with satisfactory RNA quality control and good histological morphology were sequenced for ST. Sample isolation time <15 min and abandonment of snap-frozen isopentane significantly improved RNA quality (p = 0.004, p < 0.0001) and histomorphological integrity (p = 0.02, p = 0.02). Selection of a suitable tissue RNA extraction kit was critical for RNA quality (p < 0.0001). No difference between 6 ≤ RIN <7 and RIN >7 in ST sequencing results was found, indicating that RIN ≥6 can be used as a criterion for qualified RNA quality control. Therefore, fresh tissues washed as soon as possible with cold PBS and then dried using OCT for snap freezing are currently the best method for preparing spatial sections of nose and skull base tumor tissues of different pathological types.
UNASSIGNED: This study is the first to investigate the feasibility of applying ST to different pathological types of nose and skull base tumors and to demonstrate the widespread application of ST in tumors. Rational optimization of spatial slide preparation procedures and exploration of individualized pre-sequencing protocols are used as the first stage to ensure the quality of spatial sequencing and lay the foundation for subsequent spatial analysis.
UNASSIGNED: Frozen tissue blocks from patients with squamous cell carcinoma and adenocarcinoma of the nose and skull base diagnosed at our institution were prepared. The effects of different procedures and pathological tissue types on slide quality were explored and evaluated using RNA integrity number (RIN) and HE scores as criteria. The effects of different RIN values on ST sequencing data were explored.
UNASSIGNED: A total of 43 samples were obtained from 26 patients, including 22 with squamous carcinomas and 21 with adenocarcinomas. Thirteen samples with satisfactory RNA quality control and good histological morphology were sequenced for ST. Sample isolation time <15 min and abandonment of snap-frozen isopentane significantly improved RNA quality (p = 0.004, p < 0.0001) and histomorphological integrity (p = 0.02, p = 0.02). Selection of a suitable tissue RNA extraction kit was critical for RNA quality (p < 0.0001). No difference between 6 ≤ RIN <7 and RIN >7 in ST sequencing results was found, indicating that RIN ≥6 can be used as a criterion for qualified RNA quality control. Therefore, fresh tissues washed as soon as possible with cold PBS and then dried using OCT for snap freezing are currently the best method for preparing spatial sections of nose and skull base tumor tissues of different pathological types.
UNASSIGNED: This study is the first to investigate the feasibility of applying ST to different pathological types of nose and skull base tumors and to demonstrate the widespread application of ST in tumors. Rational optimization of spatial slide preparation procedures and exploration of individualized pre-sequencing protocols are used as the first stage to ensure the quality of spatial sequencing and lay the foundation for subsequent spatial analysis.
摘要：
未经证实：空间转录组（ST）在空间水平上提供了肿瘤细胞的分子谱，给肿瘤和肿瘤微环境的研究带来了新的进展。本研究总结了我院两种不同病理类型的鼻和颅底肿瘤的空间切片制备中的经验和教训,目的是为研究人员提供指南，以避免浪费宝贵的样本，并为ST在临床实践中的应用提供依据。
UNASSIGNED：准备了在我们机构诊断的鳞状细胞癌和鼻和颅底腺癌患者的冷冻组织块。使用RNA完整性数（RIN）和HE评分作为标准，探索和评估了不同程序和病理组织类型对载玻片质量的影响。探讨了不同RIN值对ST测序数据的影响。
未经评估：共从26名患者中获得43个样本，包括22例鳞状细胞癌和21例腺癌。对13个具有令人满意的RNA质量控制和良好组织学形态的样品进行了ST测序。样品分离时间<15分钟，放弃速冻异戊烷显着提高了RNA质量（p=0.004，p<0.0001）和组织形态学完整性（p=0.02，p=0.02）。选择合适的组织RNA提取试剂盒对于RNA质量至关重要(p<0.0001)。ST测序结果6≤RIN<7和RIN>7无差异,表明RIN≥6可作为合格RNA质量控制的标准。因此,用冷PBS尽快清洗新鲜组织,然后使用OCT进行快速冷冻干燥,是目前制备不同病理类型的鼻和颅底肿瘤组织空间切片的最佳方法.
UNASSIGNED：本研究首次探讨了将ST应用于不同病理类型的鼻和颅底肿瘤的可行性，并证明了ST在肿瘤中的广泛应用。以合理优化空间载玻片制备程序和探索个体化预测序方案为第一阶段，保证空间测序质量，为后续空间分析奠定基础。
UNASSIGNED：准备了在我们机构诊断的鳞状细胞癌和鼻和颅底腺癌患者的冷冻组织块。使用RNA完整性数（RIN）和HE评分作为标准，探索和评估了不同程序和病理组织类型对载玻片质量的影响。探讨了不同RIN值对ST测序数据的影响。
未经评估：共从26名患者中获得43个样本，包括22例鳞状细胞癌和21例腺癌。对13个具有令人满意的RNA质量控制和良好组织学形态的样品进行了ST测序。样品分离时间<15分钟，放弃速冻异戊烷显着提高了RNA质量（p=0.004，p<0.0001）和组织形态学完整性（p=0.02，p=0.02）。选择合适的组织RNA提取试剂盒对于RNA质量至关重要(p<0.0001)。ST测序结果6≤RIN<7和RIN>7无差异,表明RIN≥6可作为合格RNA质量控制的标准。因此,用冷PBS尽快清洗新鲜组织,然后使用OCT进行快速冷冻干燥,是目前制备不同病理类型的鼻和颅底肿瘤组织空间切片的最佳方法.
UNASSIGNED：本研究首次探讨了将ST应用于不同病理类型的鼻和颅底肿瘤的可行性，并证明了ST在肿瘤中的广泛应用。以合理优化空间载玻片制备程序和探索个体化预测序方案为第一阶段，保证空间测序质量，为后续空间分析奠定基础。
