smRNA-FISH was performed according to the protocol of the RNAscope Multiplex Fluorescent v2 Assay (ACD-bio. 323110). In brief, embryos were fixed for 26-30h in 4%paraformaldehyde at 4°C and then dehydrated in methanol. For smRNA-FISH on sections at E6.5-E7.5, the decidua was embedded in paraffin after fixation, dehydrated in methanol and incubated for 16h in butanol at 4°C. For smRNA-FISH on sections of E9.5 chimera, the embryos were embedded in histogel (Epredia HG-4000-012) prior to paraffin embedding. Tissue sections were cut at 7 to 10 μm. Whole-mount smRNA-FISH was performed as previously described 72 (link). Probes are reported in Supplementary Table 5 . To develop probes, we have used Opal dyes from Akoya Bioscience (Opal-520, Opal-570 and Opal-650, from 1:100 to 1:200). Embryos or sections were imaged using an AxioZoom.V16 microscope (Zeiss) or an LSM800 confocal microscope (Zeiss). When analysing litters from Mesp1-Cre or Zic3 KO lines, embryos were genotyped by PCR after imaging.
Hg 4000 012
Manufactured by Epredia
The HG-4000-012 is a laboratory instrument designed for sample preparation. It is capable of performing tissue processing and dehydration tasks. The device specifications and technical details are available upon request.
Automatically generated - may contain errors
2 protocols using hg 4000 012
1
Single-Molecule RNA FISH in Mouse Embryos
2
Single-Molecule RNA FISH in Mouse Embryos
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smRNA-FISH was performed according to the protocol of the RNAscope Multiplex Fluorescent v2 Assay (ACD-bio. 323110). In brief, embryos were fixed for 26-30h in 4%paraformaldehyde at 4°C and then dehydrated in methanol. For smRNA-FISH on sections at E6.5-E7.5, the decidua was embedded in paraffin after fixation, dehydrated in methanol and incubated for 16h in butanol at 4°C. For smRNA-FISH on sections of E9.5 chimera, the embryos were embedded in histogel (Epredia HG-4000-012) prior to paraffin embedding. Tissue sections were cut at 7 to 10 μm. Whole-mount smRNA-FISH was performed as previously described 72 (link). Probes are reported in Supplementary Table 5 . To develop probes, we have used Opal dyes from Akoya Bioscience (Opal-520, Opal-570 and Opal-650, from 1:100 to 1:200). Embryos or sections were imaged using an AxioZoom.V16 microscope (Zeiss) or an LSM800 confocal microscope (Zeiss). When analysing litters from Mesp1-Cre or Zic3 KO lines, embryos were genotyped by PCR after imaging.
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