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使用免疫荧光成像和核大小定量分析果蝇肠道肿瘤细胞异质性的方案。

Protocol to analyze Drosophila intestinal tumor cellular heterogeneity using immunofluorescence imaging and nuclear size quantification.

机构信息

Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.

出版信息

STAR Protoc. 2024 Jun 21;5(2):102946. doi: 10.1016/j.xpro.2024.102946. Epub 2024 Mar 11.

DOI:10.1016/j.xpro.2024.102946
PMID:38470911
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10945268/
Abstract

Drosophila intestinal tumors show an extended cellular heterogeneity. We devise a protocol to assess tumor cell heterogeneity by employing nuclear size measurement and immunofluorescence-based cell lineage analysis. We describe steps for intestinal dissection, staining, and imaging, followed by detailed procedures for nuclear size analysis. This approach detects overall heterogeneity across the entire tumor cell population and deviations within specific cell populations. The procedure is also applicable for analyzing the heterogeneity of wild-type intestinal cells in various contexts. For complete details on the use and execution of this protocol, please refer to Pranoto et al..

摘要

果蝇肠道肿瘤表现出广泛的细胞异质性。我们设计了一种通过核大小测量和基于免疫荧光的细胞谱系分析来评估肿瘤细胞异质性的方案。我们描述了肠道解剖、染色和成像的步骤，然后详细介绍了核大小分析的步骤。这种方法可以检测整个肿瘤细胞群体的整体异质性和特定细胞群体内的偏差。该程序也适用于分析各种情况下野生型肠道细胞的异质性。有关该方案使用和执行的完整详细信息，请参阅 Pranoto 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/ca6980af96c5/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/624dd4a1ceb7/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/e35f13c958d6/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/ab6cc960bc01/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/1444fd069672/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/ca6980af96c5/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/624dd4a1ceb7/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/e35f13c958d6/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/ab6cc960bc01/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/1444fd069672/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95d/10945268/ca6980af96c5/gr4.jpg

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本文引用的文献

1
The roles of the native cell differentiation program aberrantly recapitulated in Drosophila intestinal tumors.在果蝇肠道肿瘤中异常重现的天然细胞分化程序的作用。
Cell Rep. 2023 Oct 31;42(10):113245. doi: 10.1016/j.celrep.2023.113245. Epub 2023 Oct 12.
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Remodeling of E-cadherin subcellular localization during cell dissemination.细胞扩散过程中 E-钙黏蛋白细胞内定位的重塑。
Mol Biol Cell. 2023 May 1;34(5):ar46. doi: 10.1091/mbc.E23-03-0087. Epub 2023 Mar 29.
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An image analysis method for regionally defined cellular phenotyping of the midgut.
一种用于中肠区域性定义细胞表型分析的图像分析方法。
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Wear and tear of the intestinal visceral musculature by intrinsic and extrinsic factors.内在和外在因素对肠内脏肌的磨损。
Dev Dyn. 2022 Aug;251(8):1291-1305. doi: 10.1002/dvdy.473. Epub 2022 Apr 7.
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Microbes affect gut epithelial cell composition through immune-dependent regulation of intestinal stem cell differentiation.微生物通过免疫依赖的调节肠道干细胞分化来影响肠道上皮细胞组成。
Cell Rep. 2022 Mar 29;38(13):110572. doi: 10.1016/j.celrep.2022.110572.
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Immune regulation of intestinal-stem-cell function in Drosophila.果蝇肠道干细胞功能的免疫调控。
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An amuse-bouche of stem cell regulation: Underlying principles and mechanisms from adult Drosophila intestinal stem cells.干细胞调控的开胃小菜：来自成年果蝇肠道干细胞的基本原理和机制
Curr Opin Cell Biol. 2021 Dec;73:58-68. doi: 10.1016/j.ceb.2021.05.007. Epub 2021 Jul 1.
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Tumors overcome the action of the wasting factor ImpL2 by locally elevating Wnt/Wingless.肿瘤通过局部升高 Wnt/Wingless 来克服消耗因子 ImpL2 的作用。
Proc Natl Acad Sci U S A. 2021 Jun 8;118(23). doi: 10.1073/pnas.2020120118.
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Dissemination of Ras-transformed cells requires the mechanosensitive channel Piezo.Ras 转化细胞的扩散需要机械敏感通道 Piezo。
Nat Commun. 2020 Jul 16;11(1):3568. doi: 10.1038/s41467-020-17341-y.
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