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Leica tcs sp5 x system

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The Leica TCS SP5 X is a laser scanning confocal microscope system designed for advanced imaging applications. It provides high-resolution, multi-channel imaging capabilities with a range of laser options and detection configurations.

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Lab products found in correlation

Image pro plus software, by Media Cybernetics (2 mentions) Image pro premier 9, by Media Cybernetics (1 mentions) Scn400, by Leica Microsystems (1 mentions)

5 protocols using leica tcs sp5 x system

1

Measuring Liver Fibrosis and VEGF-A

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The expression of Sct-dependent microRNA 125b (that regulates IBDM and liver fibrosis through changes in the expression of biliary VEGF-A) was measured by qPCR9 (link). By immunohistochemistry, we evaluated the semiquantitative immunoreactivity for VEGF-A in liver sections (4–5 µm thick, 10 different fields analyzed from 3 different samples). When 0–5% of bile ducts were positive for VEGF-A, a negative score was assigned; a +/− score was assigned when 6–10% of bile ducts were positive; a + score was assigned when 11–30% of bile ducts were positive; a ++ score was assigned with 31–60% of bile ducts positive; and a score +++ was assigned when more than 61% of bile ducts were positive. The expression of: (i) VEGF-A/R-2 in cholangiocytes and total liver; (ii) IL-6 and TNF-α in cholangiocytes; and (iii) CD31 and vWF in total liver was evaluated by immunoblots and/or qPCR. Immunofluorescence for the expression of CD31 was performed in frozen liver sections (10 µm thick). Immunofluorescent staining was visualized using Leica TCS SP5 X system (Leica Microsystems Inc.).
Zhou T., Wu N., Meng F., Venter J., Giang T.K., Francis H., Kyritsi K., Wu C., Franchitto A., Alvaro D., Marzioni M., Onori P., Mancinelli R., Gaudio E., Glaser S, & Alpini G. (2018). Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2−/− mice by diminishing senescence of cholangiocytes. Laboratory investigation; a journal of technical methods and pathology, 98(11), 1449-1464.
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2

Liver Fibrosis Assessment by Sirius Red and Immunofluorescence

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Liver fibrosis was assessed by Sirius red staining to quantify collagen deposition in liver sections [14 (link)]. Following Sirius red staining, slides were scanned by a digital scanner (SCN400; Leica Microsystems, Buffalo Grove, IL) and quantified by the Image-Pro Premier 9.1 (Media Cybernetics, Inc., Rockville, MD). In addition, immunofluorescence staining was performed for Col1a1 co-stained with CK-19 and desmin in frozen liver sections (8 μm). Immunofluorescent staining was visualized using Leica TCS SP5 X system (Leica Microsystems Inc.) and Olympus F300 from Texas A&M Integrated Microscopy Imaging Laboratory. We also measured the expression of Col1a1, Fn1 and TGF-β1 in isolated cholangiocytes and total liver samples by qPCR.
Zhou T., Kyritsi K., Wu N., Francis H., Yang Z., Chen L., O'Brien A., Kennedy L., Ceci L., Meadows V., Kusumanchi P., Wu C., Baiocchi L., Skill N.J., Saxena R., Sybenga A., Xie L., Liangpunsakul S., Meng F., Alpini G, & Glaser S. (2019). Knockdown of vimentin reduces mesenchymal phenotype of cholangiocytes in the Mdr2−/− mouse model of primary sclerosing cholangitis (PSC). EBioMedicine, 48, 130-142.
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3

Measuring Liver Fibrosis and VEGF-A

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The expression of Sct-dependent microRNA 125b (that regulates IBDM and liver fibrosis through changes in the expression of biliary VEGF-A) was measured by qPCR9 (link). By immunohistochemistry, we evaluated the semiquantitative immunoreactivity for VEGF-A in liver sections (4–5 µm thick, 10 different fields analyzed from 3 different samples). When 0–5% of bile ducts were positive for VEGF-A, a negative score was assigned; a +/− score was assigned when 6–10% of bile ducts were positive; a + score was assigned when 11–30% of bile ducts were positive; a ++ score was assigned with 31–60% of bile ducts positive; and a score +++ was assigned when more than 61% of bile ducts were positive. The expression of: (i) VEGF-A/R-2 in cholangiocytes and total liver; (ii) IL-6 and TNF-α in cholangiocytes; and (iii) CD31 and vWF in total liver was evaluated by immunoblots and/or qPCR. Immunofluorescence for the expression of CD31 was performed in frozen liver sections (10 µm thick). Immunofluorescent staining was visualized using Leica TCS SP5 X system (Leica Microsystems Inc.).
Zhou T., Wu N., Meng F., Venter J., Giang T.K., Francis H., Kyritsi K., Wu C., Franchitto A., Alvaro D., Marzioni M., Onori P., Mancinelli R., Gaudio E., Glaser S, & Alpini G. (2018). Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2−/− mice by diminishing senescence of cholangiocytes. Laboratory investigation; a journal of technical methods and pathology, 98(11), 1449-1464.
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4

Evaluating Liver Fibrosis via Sirius Red and Immunofluorescence

Cited 1 time
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Liver fibrosis was evaluated by Sirius Red staining in paraffin-embedded liver sections (4–5 µm thick, 10 different fields analyzed from 3 samples from 3 animals). Collagen content was quantified by Image-Pro Plus software (Media Cybernetics, Silver Springs, MD)22 (link). Immunofluorescence double staining was performed for Col1a1 (co-stained with CK-19) or α-SMA (costained with desmin) in frozen liver sections (10 µm thick). Immunofluorescent staining was visualized using Leica TCS SP5 X system (Leica Microsystems Inc.). The mRNA expression of Col1a1 and FN-1 was evaluated in cholangiocytes and/or HSCs by qPCR and/or immunoblots. TGF-β1 levels in serum and cholangiocyte supernatant were measured by ELISA kits2 (link).
Zhou T., Wu N., Meng F., Venter J., Giang T.K., Francis H., Kyritsi K., Wu C., Franchitto A., Alvaro D., Marzioni M., Onori P., Mancinelli R., Gaudio E., Glaser S, & Alpini G. (2018). Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2−/− mice by diminishing senescence of cholangiocytes. Laboratory investigation; a journal of technical methods and pathology, 98(11), 1449-1464.
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5

Evaluating Liver Fibrosis via Sirius Red and Immunofluorescence

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Liver fibrosis was evaluated by Sirius Red staining in paraffin-embedded liver sections (4–5 µm thick, 10 different fields analyzed from 3 samples from 3 animals). Collagen content was quantified by Image-Pro Plus software (Media Cybernetics, Silver Springs, MD)22 (link). Immunofluorescence double staining was performed for Col1a1 (co-stained with CK-19) or α-SMA (costained with desmin) in frozen liver sections (10 µm thick). Immunofluorescent staining was visualized using Leica TCS SP5 X system (Leica Microsystems Inc.). The mRNA expression of Col1a1 and FN-1 was evaluated in cholangiocytes and/or HSCs by qPCR and/or immunoblots. TGF-β1 levels in serum and cholangiocyte supernatant were measured by ELISA kits2 (link).
Zhou T., Wu N., Meng F., Venter J., Giang T.K., Francis H., Kyritsi K., Wu C., Franchitto A., Alvaro D., Marzioni M., Onori P., Mancinelli R., Gaudio E., Glaser S, & Alpini G. (2018). Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2−/− mice by diminishing senescence of cholangiocytes. Laboratory investigation; a journal of technical methods and pathology, 98(11), 1449-1464.
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