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在平面生物中将光现场混合与免疫光和乳素染色相结合

Daniel Font-Martín^1,2, Eudald Pascual-Carreras^1,2, Emili Saló^3,4

¹Department of Genetics, Microbiology and Statistics, Universitat de Barcelona, Barcelona, Catalunya, Spain.

Methods in molecular biology (Clifton, N.J.)

|July 10, 2023

概括

这项研究提出了一种结合光 in situ 杂交 (FISH) 和免疫光 (IF) 的新方法,用于在 Schmidtea mediterranea 中可视化基因表达和组织. 一种新的莱克固定协议增强了用于单细胞分辨率成像的信号.

科学领域:

分子生物学分子生物学
发展生物学发展生物学
细胞生物学细胞生物学

背景情况:

同时可视化Schmidtea mediterranea中的基因表达和组织组成对于理解其生物学至关重要.
光 in situ 杂交 (FISH) 和免疫光 (IF) 是此类分析的关键技术.
扩大超越FISH和IF的检测能力是非常可取的.

研究的目的:

制定一个新的FISH和IF在地中海的新组合协议.
将光结合式莱克染料整合到FISH-IF联合协议中.
为增强信号检测引入一种改进的莱克固定方法.

主要方法:

开发了一种结合FISH和IF的新方案.
在FISH-IF工作流程中纳入了光结合莱克染色.
优化了一种新的lectin固定协议,以改善信号.

主要成果:

结合的FISH-IF协议允许同时可视化分子标记物和组织结构.
莱克丁染色成功扩大了可检测的组织部件的范围.
这种新的莱克固定协议显著提高了信号强度,促进了单细胞分辨率.

结论:

开发的FISH-IF和乳素染色协议组合为 Schmidtea mediterranea 多模式分析提供了一个强大的工具.
增强的莱克固定方法改善了高分辨率成像的信号噪声比.
这种综合方法推动了对平面虫基因表达和组织动态的研究.

关键词:

抗体是一种抗体.光ISH的使用方法光结合的光剂结合的免疫光效应免疫光效应莱克是一种学菌素.平面主义者是平面主义者.

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相关概念视频

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.