结肠大体肿瘤计数、卡尺直径测量与肿瘤负荷计算
浏览实验方案对 AOM 启动、DSS 反复周期诱导的结肠炎相关结肠肿瘤模型小鼠进行肿瘤负荷终点测量：第 100 天颈椎脱臼处死，无菌分离全结肠置 15 ml 冷无菌 PBS 中，21 G 针头 5–10 ml 冷 PBS 冲洗去粪便，纵轴剖开并再次漂洗；对剖开结肠上的大体可见肿瘤逐一计数，数显卡尺测量各肿瘤直径，肿瘤负荷为单只动物全部肿瘤尺寸之和；随后拍照并以来源标注的 10% formalin solution（Sigma-Aldrich HT-501128）固定以备石蜡切片组织学分级。适用于 AOM/DSS 模型（C57BL/6 背景为主）的成瘤数量、径级分布与干预效果比较；该来源报告其饲养的 C57BL/6 背景野生型小鼠第 100 天成瘤率 100%。
适用范围与方法
样本：终点（AOM/DSS 方案约第 100 天）分离并纵向剖开的小鼠全结肠
- AOM/DSS 结肠炎相关结肠肿瘤模型（文献参考）
- 年龄
- 8–10 周龄开始 AOM/DSS 方案；终点约第 100 天肿瘤计数
- 性别
- 混合
- 组织
- colon（结肠，纵向剖开全长，大体肿瘤）
试剂与材料
实验流程
Procedure
Open the mice using sterile forceps and scissors to carefully remove the entire colon and place it in a 10 cm Petri dish with 15 ml cold sterile PBS, which should be enough to cover the entire colon.
Procedure
Carefully hold the distal part of the colon with the help of forceps and then flush to remove feces with 5-10 ml cold sterile PBS using a 21 G needle attached to a 5 ml syringe. After flushing the feces, put the cleaned colon in a new Petri dish.
Procedure
Carefully open the colon longitudinally and rinse again with cold PBS to remove remaining feces.
Procedure
For the AOM/DSS experiment, count the macroscopic tumors in the longitudinally open colon. In our in-house bred mice with mostly C57BL/6 background, tumor development is observed in 100% of the WT animals. Tumor size (average diameter) is measured using a digital caliper and tumor load per mouse is calculated by summing up the sizes of all the tumors in a given mouse. After counting tumor numbers and load, longitudinally opened colons can be photographed (Figure 1) before fixing them as ”swiss-rolls” in 10% formalin solution and then embedding in paraffin. H&E stained paraffin sections (Figure 2) should be analyzed for tumor grading and immune cells infiltration by an experience pathologist.
质量控制与输出
计数与测量在纵向剖开并漂洗去除粪便后的全结肠上进行；肿瘤负荷按每只动物全部肿瘤尺寸求和计算；大体计数后同一结肠固定并制备石蜡切片用于组织学确认。
每只动物的大体肿瘤数目、各肿瘤直径（卡尺测量）与肿瘤负荷（该动物全部肿瘤尺寸之和），配套照片与组织学分级输入；用于组间成瘤负担比较。