Tissuefaxs platform

Manufactured by TissueGnostics

Sourced in Austria

The TissueFAXS platform is a high-performance automated scanning system designed for the acquisition and analysis of microscopic images of tissue samples. It provides a standardized and efficient workflow for digitizing and processing tissue slides.

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Lab products found in correlation

Tissuequest software, by TissueGnostics (8 mentions) Strataquest, by TissueGnostics (3 mentions) Cd163, by Proteintech (1 mentions) Fitc tsa, by Wuhan Servicebio Technology (1 mentions) Cy3 tsa, by Wuhan Servicebio Technology (1 mentions) 594 tsa, by Wuhan Servicebio Technology (1 mentions) 647 tsa, by Wuhan Servicebio Technology (1 mentions) Strataquest software, by TissueGnostics (1 mentions) Gb23303, by Wuhan Servicebio Technology (1 mentions) Gb23301, by Wuhan Servicebio Technology (1 mentions) Anti cd8 antibody, by Abcam (1 mentions) Anti cd68 antibody, by Abcam (1 mentions) Anti cd4 antibody, by Abcam (1 mentions) Stempro adipogenesis differentiation kit, by Thermo Fisher Scientific (1 mentions) Stempro osteogenesis differentiation kit, by Thermo Fisher Scientific (1 mentions) Superpicture polymer detection kit, by Thermo Fisher Scientific (1 mentions) Bond max, by Leica Microsystems (1 mentions) Clone d13.14.4e, by Cell Signaling Technology (1 mentions) Histoquest software, by TissueGnostics (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)

22 protocols using tissuefaxs platform

Histological Analysis of Murine Organs

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Mice spleen, heart, intestine, liver and kidney were collected and sliced for H&E staining.²⁸ The staining procedure was performed by Bio‐Check Laboratories Ltd. (New Taipei City, Taiwan). Tissue images were acquired by TissueFAXS platform (TissueGnostics) at 200 × magnification.

Liu Y., Lin C.H., Chen K., Lai D, & Hsu F. (2023). Inactivation of EGFR/ERK/NF‐κB signalling associates with radiosensitizing effect of 18β‐glycyrrhetinic acid on progression of hepatocellular carcinoma. Journal of Cellular and Molecular Medicine, 27(11), 1539-1549.

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Tissue Cytometry and Cell-Cell Contact Quantification

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Images of the tissue specimens were acquired using the TissueFAXS platform (TissueGnostics). For quantitative analysis, the entire area of the tissue was acquired as a digital grayscale image in five channels with filter settings for FITC, Cy3, Cy5 and AF75 in addition to DAPI. Cells of a given phenotype were identified and quantitated using the TissueQuest software (TissueGnostics), with cut-off values determined relative to the positive controls. This microscopy-based multicolor tissue cytometry software permits multicolor analysis of single cells within tissue sections similar to flow cytometry. StrataQuest (TissueGnostics) software was also used to quantify cell-to-cell contact. In the StrataQuest cell-to-cell contact application, masks of the nuclei based on DAPI staining establish the inner boundary of the cytoplasm and the software “looks” outwards toward the plasma membrane boundary. The software looks for overlap of 2 cells in a 3-pixel distance from the nucleus boundary. If in this 3-pixel wide ring area, any pixel intersection of the 2 cells is required to establish a ‘‘contact’’ criterion. Although the software has been developed and validated more recently, the principle of the method and the algorithms used have been described in detail elsewhere.³⁰ (link)

Allard-Chamard H., Kaneko N., Bertocchi A., Sun N., Boucau J., Kuo H.H., Farmer J.R., Perugino C., Mahajan V.S., Murphy S.J., Premo K., Diefenbach T., Ghebremichael M., Yuen G., Kotta A., Akman Z., Lichterfeld M., Walker B.D., Yu X.G., Moriyama M., Maehara T., Nakamura S., Stone J.H., Padera R.F, & Pillai S. (2023). Extrafollicular IgD−CD27−CXCR5−CD11c− DN3 B cells infiltrate inflamed tissues in autoimmune fibrosis and in severe COVID-19. Cell Reports, 42(6), 112630.

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Multicolor Tissue Cytometry Analysis

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Images of the salivary gland specimens were acquired using the TissueFAXS platform (TissueGnostics). For quantitative analysis, the entire area of the tissue involved by the lymphoplasmacytic in-filtrate was acquired as digital grayscale images in four channels with filter settings for FITC, Cy3, and Cy5 in addition to DAPI. Cells of a given phenotype were identified and quantitated using Tissue-Quest software (TissueGnostics), with cutoff values determined relative to the positive controls. This microscopy-based multicolor tissue cytometry software permits multicolor analysis of single cells within tissue sections similar to flow cytometry. The principle of the method and the algorithms used have been described in detail elsewhere (Ecker & Steiner, 2004 (link)).

Maehara T., Mattoo H., Mahajan V.S., Murphy S.J., Yuen G.J., Ishiguro N., Ohta M., Moriyama M., Saeki T., Yamamoto H., Yamauchi M., Daccache J., Kiyoshima T., Nakamura S., Stone J.H, & Pillai S. (2018). The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo. Life Science Alliance, 1(1), e201800050.

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Multiplex Immunohistochemistry Analysis of Glioma Tissue

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We purchased the glioma tissue array from Wuhan Tanda Scientific Co., Ltd. (NGL1021), with ethics approvement. SOX10 (Mouse, 1:100, Proteintech, China), CD163 (Rabbit, 1:3,000, Proteintech, China), and CD68 (Rabbit, 1:3,000, Servicebio, China) were the primary Abs. Horseradish peroxidase-conjugated secondary antibody incubation (GB23301, GB23303, Servicebio, China) was the secondary antibody. The tyramide signal was amplified into TSA [FITC-TSA, CY3-TSA, 594-TSA, and 647-TSA (Servicebio, China)]. The stained slides were scanned using the TissueFAXS platform (TissueGnostics, Vienna, Austria). The spatial analysis of the stained cells was performed using the StrataQuest software (TissueGnostics, Vienna, Austria).

Xiao G., Wang K., Wang Z., Dai Z., Liang X., Ye W., Luo P., Zhang J., Liu Z., Cheng Q, & Peng R. (2022). Machine learning-based identification of SOX10 as an immune regulator of macrophage in gliomas. Frontiers in Immunology, 13, 1007461.

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Multicolor Immunohistochemistry Analysis

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The 5 um non-stained formalin-fixed and paraffin-embedded slides were applied for immune staining. Paraffin sections were then placed in a 70°C paraffin oven for 1 hour, followed by deparaffinized in xylene and then rehydration. Ag was recovered with citric acid buffer (pH 6.0) in the oven for 60 minutes. Inactivate endogenous peroxidase by incubation in 3% H₂O₂ for 15 minutes. After preincubation with 10% normal goat serum to block nonspecific sites for 10 minutes, sections are incubated overnight with primary antibodies including anti-CD8 antibody (Abcam, cat.189926), anti-CD4 antibody (Abcam, cat.133616), anti-Anti-Osteopontin antibody (Abcam, cat.269411), anti-CD68 antibody (Abcam,cat.213363), anti-BCA1 (Abcam, cat.246518), and antibody CD19 (Abcam, cat.134114) in a humidified chamber at 4°C. Secondary HRP-conjugated antibodies were added and incubated at room temperature for 10 minutes. For multicolored IHC analysis, fluorophore-conjugated TSA were used after incubation with secondary antibodies. After washing with PBS twice, the images were visualized using the TissueFAXS platform (TissueGnostics).

Xie L., Ning Z., Hua Y., Wang P, & Meng Z. (2023). Single-cell transcriptome analysis revealed the immune profile of PD-1 blockade in gallbladder carcinoma liver metastasis. Hepatology Communications, 7(5), e0131.

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Multicolor Tissue Cytometry Analysis

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Images of the tissue specimens were acquired using the TissueFAXS platform (TissueGnostics). For quantitative analysis, the entire area of the tissue was acquired as a digital grayscale image in five channels with filter settings for FITC, Cy3, Cy5 and AF75 in addition to DAPI. Both in the lungs and in lymph nodes, we analyzed the entire area including airspaces such as alveoli, blood vessels and lymph vessels. Cells of a given phenotype were identified and quantitated using the TissueQuest software (TissueGnostics), with cut-off values determined relative to the positive controls. This microscopy-based multicolor tissue cytometry software permits multicolor analysis of single cells within tissue sections similar to flow cytometry. StrataQuest (TissueGnostics) software was also used to quantify cell-to-cell contact. In the StrataQuest cell-to-cell contact application, masks of the nuclei based on DAPI staining establish the inner boundary of the cytoplasm and the software “looks” outwards towards the plasma membrane boundary. Overlap of at least 3 pixels of adjacent cell markers is required to establish a “contact” criterion. Although the software has been developed and validated more recently, the principle of the method and the algorithms used have been described in detail elsewhere [35 (link)].

Kaneko N., Boucau J., Kuo H.H., Perugino C., Mahajan V.S., Farmer J.R., Liu H., Diefenbach T.J., Piechocka-Trocha A., Lefteri K., Waring M.T., Premo K.R., Walker B.D., Li J.Z., Gaiha G., Yu X.G., Lichterfeld M., Padera RF J.r, & Pillai S. (2022). Temporal changes in T cell subsets and expansion of cytotoxic CD4+ T cells in the lungs in severe COVID-19. Clinical Immunology (Orlando, Fla.), 237, 108991.

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Quantitative Tissue Cytometry Analysis

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Images of the salivary gland specimens were acquired using the TissueFAXS platform (TissueGnostics). For quantitative analysis, the entire area of the tissue involved by the lymphoplasmacytic infiltrate was acquired as digital grayscale images in four channels with filter settings for FITC, Cy3, and Cy5 in addition to DAPI. Cells of a given phenotype were identified and quantitated using TissueQuest software (TissueGnostics), with cutoff values determined relative to the positive controls. This microscopy-based multicolor tissue cytometry software permits multicolor analysis of single cells within tissue sections similar to flow cytometry. The principle of the method and the algorithms used have been described in detail elsewhere (Ecker & Steiner, 2004 (link)).

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Evaluating Stem Cell Differentiation

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We used the StemPro^® Osteogenesis Differentiation Kit and StemPro^® Adipogenesis Differentiation Kit from Gibco^TM to test the osteogenic and adipogenic differentiation capabilities of cells, respectively. We tested the differentiation capabilities of FGF21 MSCs and mCherry MSCs with and without labeling with 25 µg/mL of IO-MI NPs in medium without FBS supplementation at 37°C and 5% CO₂. We followed the kit's manufacturer’s protocols to induce differentiation of cells. We tested cell differentiation into the osteogenic lineage by Alizarin Red S staining and into the adipogenic lineage by Oil Red O staining. Images were acquired using the TissueFAXS platform (TissueGnostics, Vienna, Austria) inverted microscope.

Ali A.A., Shahror R.A, & Chen K.Y. (2020). Efficient Labeling Of Mesenchymal Stem Cells For High Sensitivity Long-Term MRI Monitoring In Live Mice Brains. International Journal of Nanomedicine, 15, 97-114.

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Quantitative Immunofluorescence Analysis of Salivary Gland Tissue

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Tissue samples were obtained from whole SMGs^{14 (link)} of 16 patients with IgG4-DS (see online supplementary E4) and 6 with CS (see online supplementary table E5) and from LSGs of 15 patients with active-SS (see online supplementary table E6) and 5 healthy controls (see online supplementary table E5).
These tissue samples were fixed in formalin, embedded in paraffin and sectioned. These specimens were incubated with antibodies to transforming growth factor -β1 (TGF-β1) (clone MAB246; R&D Systems), IFN-γ (clone sc-74108; Santa Cruz Biotechnology), GZMA (clone LS-C312742; LSBio), GATA3 (clone CM405A; Biocare), CD8 (clone ab85792; abcam) and CD4 (clone CM153A; Biocare) followed by incubation with secondary antibody using a SuperPicTure Polymer Detection Kit (Invitrogen) and an Opal 3-Plex Kit (Fluorescein, Cyanine3 and Cyanine5) (Perkin Elmer). The samples were mounted with ProLong antifade-containing 4′, 6-diamidino-2-phenylindole (DAPI) (Invitrogen). Images were acquired with the TissueFAXS platform (TissueGnostics).¹² TissueFAXS is an analytical instrument that combines the advantages of multichannel microscopy and automated high-resolution imaging with the scientific accuracy of flow cytometry (http://www.bga.su/info/TissueFAXS). Stained cells were automatically counted in the salivary gland specimens.

Maehara T., Mattoo H., Ohta M., Mahajan V.S., Moriyama M., Yamauchi M., Drijvers J., Nakamura S., Stone J.H, & Pillai S.S. (2016). Lesional CD4+ IFN-γ+ cytotoxic T lymphocytes in IgG4-related dacryoadenitis and sialoadenitis. Annals of the rheumatic diseases, 76(2), 377-385.

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Immunohistochemical Analysis of Tumor Markers

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Mice were killed on day 14 after treatments. Tumors were removed from mice and fixed in 4% paraformaldehyde (PFA) at 4°C. Paraffin-embedded tumor tissues were sectioned in 5 μm-thick slices by Bio-Check Laboratories Ltd (New Taipei City, Taiwan). Protein levels in tumor tissues were evaluated with IHC staining. The instructions provided with the kit were followed to perform IHC staining. The sectioned slices were immunohistochemically stained with MMP-9, VEGF, MCL-1, XIAP, C-FLIP, Cyclin-D1, anti-P-ERK, NF-κB p65 (Ser536), P-AKT (Ser473), Caspase-9, -8, and -3 antibodies respectively. The stained slides were scanned using the microscopy-based TissueFAXS platform (TissueGnostics, Vienna, Austria) and images were captured at 100× magnification [12 (link)]. ImageJ software version 1.50 (National Institutes of Health, Bethesda, MD, U.S.A.) was used to evaluate indices of positivity on IHC slides.

Weng M.C., Wang M.H., Tsai J.J., Kuo Y.C., Liu Y.C., Hsu F.T, & Wang H.E. (2018). Regorafenib inhibits tumor progression through suppression of ERK/NF-κB activation in hepatocellular carcinoma bearing mice. Bioscience Reports, 38(3), BSR20171264.

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