微卫星不稳定性(MSI)小鼠结直肠癌模型(文献参考)
将floxed Msh2等位基因与Lynch综合征相关错义突变(Msh2 G674D)或Msh2 Δ7 null突变相结合,并在Villin-Cre转基因小鼠中实现肠上皮特异性敲除,导致肠道肿瘤发生,并产生具有MSI-high结直肠癌典型特征的化疗耐药表型。
Outcome measures
获得可供光学显微镜观察的 H&E 染色组织切片:细胞核蓝紫色、细胞质与结缔组织纤维粉红色,用于观察组织中细胞的整体结构与形态细节。本方案本身不产出病理评分数值,也不提供特定分子标志物的定量结果。
Experimental animals
微卫星不稳定性(MSI)小鼠结直肠癌模型(文献参考)
- Age
- not reported in source; ex-vivo endpoint-collected fixed tissue section, HE readout
- Tissue
- colon
微卫星不稳定性(MSI)小鼠结直肠癌模型(文献参考)
- Age
- 未记载周龄,终点取材组织切片 HE 判读(离体)
- Tissue
- colon
Experimental procedures
脱蜡(Dewaxing) 石蜡切片置于二甲苯中脱蜡 3 次,每次 00:03:00。 复水(Rehydration) 石蜡切片依次置于 100% 乙醇中 3 次、每次 00:03:00,再置于 95% 乙醇中 1 次、00:03:00,随后用水漂洗。 苏木素核染色(Hematoxylin staining) 石蜡切片以苏木素染液染色 00:00:20,随后在流水下冲洗 00:01:00。 分化与返蓝(Differentiation and bluing) 石蜡切片置于返蓝液中 00:01:00,随后在流水下冲洗 00:01:00;此步使细胞核呈现蓝色。 伊红染色(Eosin staining) 石蜡切片以伊红复染 00:00:10;此步使细胞质成分与结缔组织纤维呈现粉红色。 脱水与透明(Dehydration and Clearing) 石蜡切片依次置于 95% 乙醇中 00:02:00、100% 乙醇中 2 次、每次 00:02:00,随后置于二甲苯中 2 次、每次 00:02:00。 封片(Mounting) 石蜡切片以 Permount 封片介质加盖玻片封固。
Model identity
- Model name
- 微卫星不稳定性(MSI)小鼠结直肠癌模型(文献参考)
- Species
- mouse
- Taxonomy ID
- 10090
- Genotype / zygosity
- Msh2 floxed; Villin-Cre
- Allele
- Msh2 floxed
Biological characteristics
- Research questions
- 综述归纳VCMsh2 LoxP与Villin-Cre组合模型在模拟Lynch综合征及MSI-high结直肠癌化疗耐药特征中的应用价值。
- Limitations
- 文献参考模型,不代表公司实际保有、供应或质检。本包内无原研实际使用,依据为综述归纳。模型局限:该模型为条件性MMR缺陷模型,依赖于Cre/LoxP系统实现肠上皮特异性失活,其背景突变组合(G674D或Δ7)与人群Lynch综合征的遗传异质性存在差异;本包内仅以综述形式推荐,尚未见原研直接采用该模型开展实验。
Experimental methods
Experimental protocols
- Hematoxylin and Eosin (H&E) staining for histopathology
Hematoxylin and eosin (H&E) staining of fixed, paraffin-embedded tissue sections (4 μm). The workflow covers dewaxing, rehydration, hematoxylin nuclear staining, bluing, eosin counterstaining, dehydration and clearing, and coverslipping, yielding blue-purple nuclei and pink cytoplasm and connective tissue fibers for light-microscopic assessment of cellular and tissue morphology.
Reagents and materials
- 组织学二甲苯(混合异构体)
脱蜡剂
- Ethanol
复水梯度溶剂(去除二甲苯)
- 95% 乙醇溶液
复水梯度溶剂
- 自来水(组织学染色用)
复水后漂洗用水(后续各流水冲洗共用)
- Epredia Signature Series 苏木素染液(7221)
细胞核染色
- Epredia Signature Series 返蓝液(7301)
返蓝(使细胞核转蓝)
- Epredia Signature Series 伊红 Y 染液(7111)
细胞质与结缔组织纤维复染
- Permount 组织学封片剂(Fisher SP15)
封片介质
References
- Bürtin Florian, Mullins Christina S, Linnebacher Michael. Mouse models of colorectal cancer: Past, present and future perspectives. World journal of gastroenterology. 2020; PMID: 32308343
- Nathalia L. Oliveira; Samantha Gruenheid. Hematoxylin and Eosin (H&E) staining for histopathology. protocols.io, v1, 2026-07-31. DOI: 10.17504/protocols.io.kxygxr9jdg8j
- Epredia Signature Series Hematoxylin (7221), 产品信息页, Fisher Scientific
- Epredia Signature Series Bluing Reagent (7301), 产品信息页, Fisher Scientific
- Epredia Signature Series Eosin Y (7111), 产品信息页, Fisher Scientific
- Xylene, Fisherbrand HistoPrep (HC7001GAL), 产品信息页, Fisher Scientific
- Fisher Chemical Permount Mounting Medium (SP15 产品族), 产品信息页, Fisher Scientific
- Ethanol (CID 702, CAS 64-17-5), PubChem 化合物标准记录
- ARRIVE 2.0 guidelines