Four-micrometer sections of formalin-fixed and paraffin-embedded liver biopsy specimens were deparaffinized with xylene and rehydrated with decreasing concentrations of ethanol in water. Antigen retrieval was achieved in sodium citrate buffer (pH 6.0) for 20 min at 95 °C. Tissues were stained by immunohistochemistry with either anti-CD8a antibody (1:5000, Ebioscience, ThermoFisher Scientific, San Diego, CA, USA, clone 4SM15, catalog number: 14-0808-82), anti-CD4 (1:400, Cell signaling, Danvers, MA, USA, clone D7D22, catalog number: #25229), anti-CD3 (1:1000, Dako, Agilent, Santa Clara, CA, USA, clone A0452, catalog number: 790-4341), or anti-Ki67 antibody (1:150, Millipore, Merk, Darmstadt, Germany, catalog number: AB9260). The reaction was revealed using EnVision™+System-HRP (Dako, Agilent, catalog number: K4003) or DAB Substrate Kit (Abcam, Cambridge, UK, catalog number: ab64238), and sections were counterstained with hematoxylin. Images were acquired using a panoramic 250 Flash III digital slide scanner (3DHistech, Budapest, Hungary) or with AxioScan.Z1 (Carl Zeiss Microscopy GmbH, Jena, Germany) with a 20× objective and analyzed using Definiens System (Definiens AG, Munich, Germany) or ZEN2 blue edition software (Carl Zeiss Microscopy GmbH, Jena, Germany).
250 flash 3 digital slide scanner
Manufactured by 3DHISTECH
Sourced in Hungary
The 250 Flash III is a digital slide scanner designed for high-throughput scanning of glass microscope slides. It captures high-resolution digital images of tissue samples with a maximum scan area of 15 mm x 15 mm. The scanner utilizes a CMOS camera sensor and can capture images at a maximum resolution of 0.25 μm/pixel.
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Lab products found in correlation
Anti ki 67 antibody, by Merck Group (1 mentions)
Dab substrate kit, by Abcam (1 mentions)
Anti cd4, by Cell Signaling Technology (1 mentions)
Axioscan z1, by 3DHISTECH (1 mentions)
Zen 2 blue edition software, by Zeiss (1 mentions)
Envision system hrp, by Agilent Technologies (1 mentions)
Mowiol mounting medium, by Merck Group (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 secondary antibody, by Thermo Fisher Scientific (1 mentions)
2 protocols using 250 flash 3 digital slide scanner
1
Immunohistochemical Analysis of Liver Biopsy
2
Immunofluorescence Staining of Human Skin
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O.C.T.-embedded human healthy skin samples were snap-frozen in liquid nitrogen, and 8-µm cryostat sections were prepared. After fixation with 4% PFA for 10 min at RT, sections were blocked with PBS containing 5% donkey serum, 0.1% Triton X-100, and 1% BSA for 1 h at RT, followed by incubation with mouse anti-human podoplanin (Agilent; clone D2-40) and polyclonal rabbit anti-human LYVE-1 (Reliatech) antibodies overnight at 4°C. After abundant washes with PBS, sections were incubated with donkey anti-mouse Alexa Fluor 647 and donkey anti-rabbit Alexa Fluor 488 secondary antibodies (Thermo Fisher Scientific) together with Hoechst 33342 (Thermo Fisher Scientific) for 30 min at RT. After abundant washes with PBS, sections were incubated with anti-human αSMA-Cy3 antibody (Sigma-Aldrich) for 2 h at RT. Sections were washed with PBS and mounted with Mowiol mounting medium (Sigma-Aldrich). Images were acquired using a panoramic 250 Flash III digital Slide Scanner (3D Histech). Human studies were approved by the local ethics committee (Kantonale Ethikkommission Zürich, approval 2017–00687), and informed consent was obtained from participants.
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