Combined single-molecule fluorescence in situ hybridization and immunohistochemistry analysis in intact murine dorsal root ganglia and sciatic nerve.
Combined single-molecule fluorescence in situ hybridization and immunohistochemistry analysis in intact murine dorsal root ganglia and sciatic nerve.
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DOI：
10.1016/j.xpro.2021.100555
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发表时间：
2021-06-18
期刊：
影响因子：
--
通讯作者：
Meffert MK
中科院分区：
文献类型：
--
作者：
Li X;Eadara S;Jeon S;Liu Y;Muwanga G;Qu L;Caterina MJ;Meffert MK
单分子荧光原位杂交技术(SmFISH)可用于基因表达的空间定位。该方案展示了在完整的小鼠感觉神经系统组织中smFISH的保真度和灵活性方面的进展。使用RNAScope探针的方法允许多路传输、增强靶标特异性和免疫组织化学兼容性。计算策略提高了背根神经节中成簇转录的点扩散函数和混合的坐骨神经细胞类型的3D掩蔽的mRNA斑点的量化准确性。方法在神经元中对中等(Lin28a)和中等(Ppib)稳态丰度的mRNAs进行了验证。小鼠周围神经系统单基因表达的免疫组织化学定位点扩散函数(PSF)分析方法用于基因转录定量的三维掩蔽策略单分子荧光原位杂交法(SmFISH)可用于基因表达的空间定位。该方案展示了在完整的小鼠感觉神经系统组织中smFISH的保真度和灵活性方面的进展。使用RNAScope探针的方法允许多路传输、增强靶标特异性和免疫组织化学兼容性。计算策略提高了背根神经节中成簇转录的点扩散函数和混合的坐骨神经细胞类型的3D掩蔽的mRNA斑点的量化准确性。方法在神经元中对中等(Lin28a)和中等(Ppib)稳态丰度的mRNAs进行了验证。
Single-molecule fluorescence in situ hybridization (smFISH) allows spatial mapping of gene expression. This protocol presents advances in smFISH fidelity and flexibility in intact murine sensory nervous system tissue. An approach using RNAscope probes allows multiplexing, enhanced target specificity, and immunohistochemistry compatibility. Computational strategies increase quantification accuracy of mRNA puncta with a point spread function for clustered transcripts in the dorsal root ganglion and 3D masking for intermingled sciatic nerve cell types. Approaches are validated for mRNAs of modest (Lin28a) and medium (Ppib) steady-state abundance in neurons. Single mRNA spatial mapping with immunohistochemistry in murine peripheral nervous system Point spread function (PSF) analysis approach for clustered transcript quantification 3D masking strategy for mRNA quantification in intermingled sciatic nerve cell types Single-molecule fluorescence in situ hybridization (smFISH) allows spatial mapping of gene expression. This protocol presents advances in smFISH fidelity and flexibility in intact murine sensory nervous system tissue. An approach using RNAscope probes allows multiplexing, enhanced target specificity, and immunohistochemistry compatibility. Computational strategies increase quantification accuracy of mRNA puncta with a point spread function for clustered transcripts in the dorsal root ganglion and 3D masking for intermingled sciatic nerve cell types. Approaches are validated for mRNAs of modest (Lin28a) and medium (Ppib) steady-state abundance in neurons.
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影响因子：
3.7
作者：
Hildyard JCW;Rawson F;Wells DJ;Piercy RJ
通讯作者：
Piercy RJ
DOI：
10.3791/3564
发表时间：
2012-07-30
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Gage, Gregory J;Kipke, Daryl R;Shain, William
通讯作者：
Shain, William
影响因子：
10.5
作者：
Elbarbary, Reyad A.;Li, Wencheng;Maquat, Lynne E.
通讯作者：
Maquat, Lynne E.
影响因子：
4.1
作者：
Wang, Fay;Flanagan, John;Luo, Yuling
通讯作者：
Luo, Yuling
影响因子：
16
作者：
Chen LL;Carmichael GG
通讯作者：
Carmichael GG