E15.5 embryos were processed whole, while E18.5 embryos were skinned and trisected to capture the region below the mandible and above the abdomen. Samples were fixed in Bouin’s fixative (Sigma, HT101128) at room temperature and graded into EtOH, histoclear (National Diagnostics, HS-200), and paraffin before being embedded in paraffin. Coronal serial sections were cut at a thickness of 7 μm and subsequently stained with hematoxylin and eosin. Imaging was performed on a Zeiss Imager.Z2 and 3D reconstructed models were produced using Avizo (ThermoFisher): sections were aligned using the “align slices” function, lumens of the pharynx, larynx, trachea, bronchi, and esophagus model labels were selected by thresholding for unstained regions and selecting with the “magic wand” tool within the segmentation pane, model labels were processed using the “resample” tool with “voxel averaging” of “3 × 3 × 1”, a surface model was generating using “generate surface” with “constrained smoothing” of “extent” 2, and models were visualized with the “surface view” module.
Ht101128
Manufactured by Merck Group
The HT101128 is a piece of laboratory equipment designed for general use in research and diagnostic settings. It serves as a tool for various scientific applications, without further details on its specific intended use.
Automatically generated - may contain errors
3 protocols using ht101128
1
Embryonic Airway Reconstruction from Histological Sections
2
Trichrome Staining of Cardiac Collagen
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The trichrome method was used for detection of collagen fibers in the heart muscle preparations as described by Gomori [24 (link)]. Using this method, cell nuclei are stained in dark blue or black, muscle fibers are stained in red, while the connective tissue, primarily collagen, is presented in green. Based on Gomorie trichrome staining, collagen was determined in heart ventricles. Slides were fixed with 37% formaldehyde, incubated with Bouin’s reagent (Sigma-Aldrich HT-10-1-128, 15 min, 56 °C,) and, after washing with tap water, stained with Weigert’s iron hematoxylin (Sigma-Aldrich HT10-7, HT10-9). After that, slides were stained with LG (Gomori one-step trichrome [light green] procedure) reagent (Sigma-Aldrich HT10-3-16) then washed for 1 min in 0.5% acetic acid, dehydrated (50–100% ethyl alcohol) and mounted in DPX mounting medium for histology (Sigma-Aldrich). Slides were analyzed with Nicon Eclipse 80i, 20 × objective; five random fields were photographed per slide.
3
Embryonic Airway Reconstruction from Histological Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
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E15.5 embryos were processed whole, while E18.5 embryos were skinned and trisected to capture the region below the mandible and above the abdomen. Samples were fixed in Bouin’s fixative (Sigma, HT101128) at room temperature and graded into EtOH, histoclear (National Diagnostics, HS-200), and paraffin before being embedded in paraffin. Coronal serial sections were cut at a thickness of 7 μm and subsequently stained with hematoxylin and eosin. Imaging was performed on a Zeiss Imager.Z2 and 3D reconstructed models were produced using Avizo (ThermoFisher): sections were aligned using the “align slices” function, lumens of the pharynx, larynx, trachea, bronchi, and esophagus model labels were selected by thresholding for unstained regions and selecting with the “magic wand” tool within the segmentation pane, model labels were processed using the “resample” tool with “voxel averaging” of “3 × 3 × 1”, a surface model was generating using “generate surface” with “constrained smoothing” of “extent” 2, and models were visualized with the “surface view” module.
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