使用改良的瑞士滚转技术增强小鼠胃肌层空间映射
November 25th, 2025
该方案概述了一种基于最初为小鼠肠道开发的“瑞士卷”技术改进的全胃免疫染色方法。
我们的研究重点是改进轻度胃部影像的准备和分析。我们采用了最初为小鼠肠道开发的瑞士滚转技术，使所有主要胃区，包括眼底、胃体和窦，都能嵌入在同一个冷冻切片中。这使我们能够并列可视化胃的结构和细胞组成，同时保持准确的空间方向和高质量的图像。
胃是一个复杂的器官，组织脆弱。细微的错误可能损伤细胞或扭曲排列，使得高分辨率成像和免疫染色不一致。我们的瑞士滚轮技术通过将包括十二指肠和食管在内的所有区域保持在同一个冷冻块中完整，从而使实验更可靠、更可重复。
首先，用剪刀沿着十二指肠腹侧肠系膜和胃部切到血体的针点。使用装有PBS的转移移液管，冲洗胃腔内残留的食物。轻轻拉伸张开的腹部，使其保持平坦。
短暂固定后，再次将组织压平。用剪刀沿着虚线轮廓修剪腹部，形成适合滚动的均匀形状。将修剪好的胃组织放在浸有PBS的滤纸上。
将纸巾卷成瑞士卷状，并用牙签插入中心固定。卷好后，插入一根30号针以固定形状。然后将针头弯曲成90度，以稳定滚动的腹部。
将卷起的组织浸泡在4%对甲醛中，4摄氏度，固定六小时。然后轻轻取出稳定针。将组织嵌入最佳切割温度的胶片中，使用一次性基模。
然后把霉菌冻在干冰上。完全冷冻后，在零下80摄氏度下储存模具。新方法使得将眼底、胆体、窦、部分食管和十二指肠纳入单一小鼠胃瑞士翻滚切片。
苏木精和伊红染色了胃的瑞士卷，显示眼底、体、窦、食管和十二指肠结构保存良好，黏膜、黏膜下和肌肉层均清晰可见。免疫荧光染色显示卡哈尔间质细胞中表达了KIT和ANO1，肠道神经元中表达了β-微管蛋白3，确认了胃瑞士卷中的细胞保存。胃黏膜中的paneth细胞和杯状细胞中也检测到KIT表达，而ANO1在眼底柱状上皮细胞中表达较高。
β微管蛋白3的表达在胃、内分泌和分泌黏液细胞以及肠道神经元中均有表现。肌球蛋白重链11染色确认胃黏膜中存在平滑肌细胞和肌纤维母细胞，E-卡德蛋白染色显示上皮细胞，KIT 染色再次显示卡哈尔间质细胞。瑞士路技术可以推动对胃组织区域特异性变化的研究。
与传统方法分别嵌入区域不同，瑞士滚筒技术在相同条件下处理所有胃区，节省时间和试剂。这使得它成为一个实用且可扩展的平台，用于全面分析健康和疾病中的胃组织。
概述
本协议概述了一种由最初为小鼠肠道开发的“瑞士卷”技术改良而来的全胃免疫染色方法。该方法允许在保持空间定向和图像质量的同时，观察胃结构和细胞组成。
关键研究组成部分
科学领域
- 神经科学
- 胃肠病学
- 组织学
背景
- 胃是一个具有精细组织的复杂器官。
- 传统方法常常导致成像和染色的不一致。
- 瑞士卷技术允许多个胃区域一起嵌入。
- 这提高了实验的可靠性和可重复性。
研究目的
- 改善胃组织的制备和成像分析。
- 在单个冷冻切片中可视化所有主要胃区域。
- 在成像过程中保持准确的空间定向。
使用的方法
- 瑞士卷技术的适应用于全胃免疫染色。
- 使用剪刀和移液器进行组织准备。
- 将组织嵌入最佳切割温度化合物中。
- 使用苏木精和伊红染色评估组织结构。
主要结果
- 各个胃区域的结构保存良好。
- 成功地对特定细胞类型进行免疫荧光染色。
- 证实了胃组织中关键蛋白的表达。
- 确认了平滑肌细胞和上皮细胞的存在。
结论
- 瑞士卷技术增强了胃组织分析。
- 它允许同时处理所有胃区域。
- 该方法对健康和疾病研究具有实用性和可扩展性。
常见问题
Executive Industry Relevance
Consistent spatial mapping of gastric muscle layers is critical for high-confidence target validation and mechanistic de-risking in gastrointestinal drug discovery. The modified Swiss roll technique enables reproducible, side-by-side analysis of all major mouse gastric regions, supporting robust comparative studies and minimizing experimental variability. This scalable approach enhances predictive confidence for early discovery and translational research pipelines focused on gastric biology.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables comprehensive interrogation of region-specific cellular markers across the fundus, corpus, and antrum.
- Supports biological de-risking by preserving native tissue architecture for accurate pathway analysis.
- Facilitates functional target validation through high-quality immunofluorescence and histological readouts.
- Improves predictive confidence for prioritizing gastric targets in discovery portfolios.
Screening & Assay Development
- Provides standardized, reproducible tissue sections for downstream assay development and screening workflows.
- Ensures quantitative and qualitative consistency in immunostaining outputs across all gastric regions.
- Enables scalable preparation of validated biological samples for high-throughput analysis.
- Reduces reagent use and experimental time by processing all regions under identical conditions.
Translational & Preclinical Research
- Aligns spatial omics and biomarker studies with disease-relevant gastric regions for translational continuity.
- Supports risk-adjusted advancement decisions by minimizing tissue loss and experimental variability.
- Facilitates side-by-side comparison of physiological and pathological states in preclinical models.
- Enhances mechanistic de-risking for gastric disease research and therapeutic development.
Pipeline & Workflow Integration
This method integrates into the discovery continuum from early target validation through assay development and preclinical research, providing a reproducible platform for spatial and molecular analysis of gastric tissues.
- Discovery Biology: Supports hypothesis testing and pathway clarification by enabling region-specific marker analysis in native tissue context.
- Screening: Delivers assay-ready, reproducible sections for quantitative immunostaining and comparative studies.
- Analytics: Provides high-resolution, multiplexed readouts for robust statistical comparison of gastric regions.
- Translational Research: Maintains anatomical and molecular continuity from discovery through preclinical validation.
- Enterprise Reuse: Offers a scalable, standardized tissue preparation platform for diverse gastric research applications.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in gastric target studies.
- Operational Value: Enhances standardization, reproducibility, and scalability of tissue preparation workflows.
- Strategic Value: Improves go/no-go decision quality and capital efficiency by minimizing experimental variability.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of gastric research programs.
Implementation Considerations
- Requires expertise in tissue handling, fixation, and cryosectioning for optimal results.
- Needs access to cryostat, immunofluorescence, and histological imaging infrastructure.
- Demands cross-team standardization of tissue orientation and staining protocols.
- Adaptable to various mouse models and compatible with spatial omics workflows.
- Dependent on careful handling to preserve delicate gastric architecture and minimize tissue loss.
