Liver tissues were fixed in 10% formalin, followed by dehydration in a LEICA ASP300S automated vacuum tissue processor (LEICA Microsystems, Wetzlar, Germany). Following the manufacturer’s instructions, they were then paraffin-embedded and sliced at 4 μm thickness for H&E and Sirius red staining. Histopathological changes and fibrosis stage were assessed using H&E and Sirius red staining. Sirius red staining was also quantified using Leica LAS Image Analysis.
Las image analysis
Manufactured by Leica
LAS Image Analysis is a software suite designed for advanced image analysis and processing. It provides a range of tools for visualizing, measuring, and analyzing digital images, including those captured with Leica microscopes. The software offers a comprehensive set of image analysis functions to support scientific research and applications.
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Lab products found in correlation
3 protocols using las image analysis
1
Histological Analysis of Liver Fibrosis
2
Histological Assessment of Tissue Fibrosis
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Formalin-fixed paraffin-embedded tissue samples were cut into 4-µm sections, and then adhered to silanized slides. The samples were then stained by H&E and Sirius red staining to evaluate histopathological features and fibrosis stage according to manufacturer's instructions. Sirius red-positive area was further calculated by Leica LAS Image Analysis, percentage of positive staining area = positive staining area/total area × 100%.
3
RT-qPCR Analysis of Pten and Gapdh Genes
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Primer sequences 5 0 to 3 0
Pten Forward: 5 0 -CTGCAGAGTTGCACAGTATC-3 0 Pten Reverse: 5 0 -GCTGCAACATGATTGTCATC-3 0 Gapdh Forward: 5 0 -TGACCACAGTCCATGCCATC-3 0 Gapdh Reverse: 5 0 -GACGGACACATTGGGGGTAG-3 0
Abbreviation: RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
phosphate-buffered saline (PBS). Specimens were then incubated with goat anti-rabbit IgG (HþL) highly cross-adsorbed secondary antibody for 1 hour at room temperature. After washed with PBS, sections were evaluated under a Leica DM4000 biological microscope equipped with epifluorescent illumination. Raw images were analyzed using Leica LAS Image Analysis by quantifying the average intensities. 29
Pten Forward: 5 0 -CTGCAGAGTTGCACAGTATC-3 0 Pten Reverse: 5 0 -GCTGCAACATGATTGTCATC-3 0 Gapdh Forward: 5 0 -TGACCACAGTCCATGCCATC-3 0 Gapdh Reverse: 5 0 -GACGGACACATTGGGGGTAG-3 0
Abbreviation: RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
phosphate-buffered saline (PBS). Specimens were then incubated with goat anti-rabbit IgG (HþL) highly cross-adsorbed secondary antibody for 1 hour at room temperature. After washed with PBS, sections were evaluated under a Leica DM4000 biological microscope equipped with epifluorescent illumination. Raw images were analyzed using Leica LAS Image Analysis by quantifying the average intensities. 29
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