Paraffin blocks of tumor tissues were sliced into 4 µm thick slides and heated at 60°C for 1 hour. The slides were dewaxed with xylene and stained with Leica Bond Rx Automated Stainer (Leica Biosystems). Afterwards, the slides were baked for 30 min, dewaxed, placed in Bond Epitope Retrieval 2 (Leica Biosystems) and Bond Epitope Retrieval 1 (Leica Biosystems) sequentially. The slides were incubated with the PD-L1 primary antibodies (13 684S) and CD8 (98 941S) and detected using the corresponding polymer horseradish peroxidase. Visualization was performed with using the Opal TSA Plus dye 690 and 720, respectively (Akoya Biosciences). Anti-CD8 antibody was incubated after incubation with the Opal TSA-DIG reagent (Perkin-Elmer). The nuclei were stained using DAPI (4′,6-diamidino-2-phenylindole) before the slides were covered using HIGHDEF IHC fluoromount (Enzo). The images were gained with PerkinElmer Vectra V.3.0 Automated Quantitative Pathology Imaging System (Perkin-Elmer) and evaluated with the InForm software V.2.2 and TIBCO Spotfire (Perkin-Elmer).
Vectra v 3.0 automated quantitative pathology imaging system
Manufactured by PerkinElmer
Sourced in Japan
The Vectra V.3.0 Automated Quantitative Pathology Imaging System is a laboratory equipment product designed for high-throughput imaging and analysis of tissue samples. It provides automated, quantitative assessment of multiple biomarkers within a single tissue sample.
Automatically generated - may contain errors
Lab products found in correlation
Bond epitope retrieval 2, by Leica (1 mentions)
Bond epitope retrieval 1, by Leica (1 mentions)
Highdef ihc fluoromount, by Enzo Life Sciences (1 mentions)
Tibco spotfire, by PerkinElmer (1 mentions)
Bond rx automated stainer, by Leica (1 mentions)
Type 1 collagen, by BD (1 mentions)
Matrigel, by BD (1 mentions)
Diff quik staining kit, by Sysmex (1 mentions)
Transwell membrane insert, by Corning (1 mentions)
2 protocols using vectra v 3.0 automated quantitative pathology imaging system
1
Multiplex IHC for PD-L1 and CD8 Analysis
2
Quantifying TNBC Cell Invasion
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Twenty-four-well Transwell membrane inserts (diameter: 6.5 mm, pore size: 8 µm; Corning, Tewksbury, MA, USA) were coated with 10 µL of type I collagen (0.5 mg/mL, BD Biosciences, San Diego, CA, USA) and 20 µL of a 1:20 mixture of Matrigel (BD Biosciences) in PBS. After treatment with the indicated compounds for 24 h, MDA-MB-231 human TNBC cells (parent or docetaxel resistant) were harvested, resuspended in serum-free medium, and plated (2 × 105 cells/chamber) in the upper chamber of the Matrigel-coated Transwell insert. Media containing 30% FBS were used as the chemoattractant in the lower chamber. After 24 h incubation, cells that had invaded outer surfaces of lower chambers were fixed and stained using the Diff-Quik Staining Kit (Sysmex, Kobe, Japan) and imaged using the Vectra v3.0 Automated Quantitative Pathology Imaging System (Perkin Elmer, Waltham, MA, USA). Representative images from three separate experiments were evaluated, and the number of invasive cells was semi-quantified using the ImageJ v1.52a software (National Institutes of Health, Bethesda, MD, USA) [28 (link)].
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