Vectra 3 quantitative pathology imaging system

Manufactured by PerkinElmer

Sourced in United States

The VECTRA 3 Quantitative Pathology Imaging System is a digital pathology platform designed for high-resolution, multiplexed tissue imaging and analysis. The system combines advanced optics, automated slide handling, and specialized image analysis software to enable quantitative assessment of protein expression and tissue morphology in tissue samples.

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Lab products found in correlation

Phenochart software, by PerkinElmer (2 mentions) S3022, by Agilent Technologies (1 mentions) Opal 7 color fluorescent ihc kit, by PerkinElmer (1 mentions) Phenochart, by PerkinElmer (1 mentions) Inform software, by PerkinElmer (1 mentions)

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Vectra 3 (61 citations) Vectra 3.0 Automated Quantitative Pathology Imaging System (32 citations) Vectra 3 Automated Quantitative Pathology Imaging System (17 citations) Vectra imaging system (14 citations) Quantitative Pathology Imaging System (4 citations) Vectra 3.0.5 Automated Quantitative Pathology Imaging system (2 citations) Vectra V.3.0 Automated Quantitative Pathology Imaging System (2 citations)

5 protocols using vectra 3 quantitative pathology imaging system

Multiplex Immunohistochemistry of IPF Lungs

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FFPE tissue sections were first stained with Masson's trichrome to assess extent of fibrosis (supplementary figure S2A). Tissue sections from three histologically normal NDC donors and three patients with IPF whose tissue showed distinct differences between the extent of fibrosis in the lung apex compared to the lung base were used for multiplex staining. A total of nine samples were used in the analysis.
For multiplex staining, tyramide signal amplification was performed. To assess potential crossover staining, tissue was stained with individual markers during optimisation of the panel (supplementary figure S2B).
Sections were scanned using the Vectra 3 Quantitative Pathology Imaging System (Perkin Elmer, Hopkinton, MA, USA) and regions of interest were identified for subsequent analysis at 20× magnification (supplementary figure S2C). Quantification of marker positive cells was performed using HALO imaging processing software on a total of 1101 images. Data are presented as percentage of positive cells per total number of cells counted.

Jaffar J., Wong M., Fishbein G.A., Alhamdoosh M., McMillan L., Gamell-Fulla C., Ng M., Wilson N., Symons K., Glaspole I, & Westall G. (2022). Matrix metalloproteinase-7 is increased in lung bases but not apices in idiopathic pulmonary fibrosis. ERJ Open Research, 8(4), 00191-2022.

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Multispectral Imaging and Automated Immune Cell Phenotyping

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Imaging was performed using the VECTRA 3 Quantitative Pathology Imaging System (PerkinElmer) and Vectra V3.0.4 software (PerkinElmer®, Hopkinton, MA, USA). All standard epi-fluorescent filters were used; DAPI, FITC, CY3, Texas Red, and CY5. Whole slide scans were acquired using x4 magnification and subsequently scanned at x20 magnification for the multispectral regions of interest (Figure 1B). Images were first processed using the inForm software (V.2.4.8, Akoya Biosciences, MA, USA, RRID: SCR_019155) for cell and tissue segmentation and then processed through an in-house AI pipeline to phenotype immune cells, which were thereafter analyzed in FlowJo™ (Ashland, OR, USA, RRID: SCR_008520) as previously described (35 (link), 36 (link)).

Graham Martínez C., Barella Y., Kus Öztürk S., Ansems M., Gorris M.A., van Vliet S., Marijnen C.A, & Nagtegaal I.D. (2022). The immune microenvironment landscape shows treatment-specific differences in rectal cancer patients. Frontiers in Immunology, 13, 1011498.

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Multiplexed Immunohistochemistry for Immune Cell Analysis in Colorectal Cancer

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Analysis of infiltrating immune cells using multiplexed immunohistochemistry and multispectral imaging has been described previously [15 (link)]. In brief, the TMA slides were sequentially stained with antibodies against CD66b, CD8, CD20, FoxP3, CD68 and pan-Cytokeratin. The VECTRA 3 Quantitative Pathology Imaging System (PerkinElmer) was used for multispectral imaging. All standard epi-flourescent filters were used; DAPI, FITC, CY3, Texas Red, and CY5. Whole slide scans were acquired using × 10 magnification. The Phenochart software (PerkinElmer) was subsequently used to mark TMA cores from the whole slide scans for subsequent multispectral imaging using x20 magnification. Images were quantified using the InForm software in two steps. Firstly, 15 TMA cores representing the heterogeneous nature of CRC, were selected to train machine-learning algorithms for tissue segmentation, cell segmentation and cell phenotyping. These were then applied to all TMA cores. Each scanned image was examined by one observer under the supervision of an experienced gastropathologist.

Gunnarsson U., Strigård K., Edin S., Gkekas I., Mustonen H., Kaprio T., Böckelman C., Hagström J., Palmqvist R, & Haglund C. (2020). Association between local immune cell infiltration, mismatch repair status and systemic inflammatory response in colorectal cancer. Journal of Translational Medicine, 18, 178.

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Multiparametric Immunofluorescence Staining

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Rabbit monoclonal antibodies to CD4 (Cell Marque, Rocklin, CA) clone 104R-1, 1/100, high pH retrieval), mouse monoclonal antibody to FOXP3 (Abcam, Cambridge, UK), clone IgG1, 1/1000, high pH retrieval), mouse polyclonal antibody to PD-1 (Abcam, IgG1 Nat105, 1/500, low pH retrieval), mouse monoclonal antibody to CTLA-4 (MyBiosource, San Diego, CA) IgG2a/k, 1/100, high pH retrieval) were used. Antibodies were diluted using a background-reducing antibody diluent buffer S3022 (Agilent, Santa Clara, CA). A horseradish hydrogen peroxidase (HRP) linked anti-mouse and anti-rabbit secondary antibody, EnvisionTM HRP (Agilent), was used for each primary antibody species according to the manufacturer’s recommendation. Immunofluorescent signal was visualized using the TSA amplification system, OPAL™ 7-color fluorescent IHC kit (Perkin Elmer), TSA dyes 540, 620, 650, and 690 (1:50) for ten minutes, counterstained with Spectral DAPI. All slides were imaged on the Vectra® 3 Quantitative Pathology Imaging System (Perkin Elmer). Images were then examined using color separation and inForm® Software v2.1 (Perkin Elmer). All slides were scanned on the Vectra at 10× magnification in order to select for high-powered imaging at 20× (resolution of 0.5 μm per pixel) using Phenochart (Perkin Elmer).

Richardson Z.A., Deleage C., Tutuka C.S., Walkiewicz M., Del Río-Estrada P.M., Pascoe R.D., Evans V.A., Reyesteran G., Gonzales M., Roberts-Thomson S., González-Navarro M., Torres-Ruiz F., Estes J.D., Lewin S.R, & Cameron P.U. (2021). Multiparameter immunohistochemistry analysis of HIV DNA, RNA and immune checkpoints in lymph node tissue. Journal of immunological methods, 501, 113198.

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Multispectral Imaging of TMAs

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Imaging was performed using the VECTRA 3 Quantitative Pathology Imaging System (PerkinElmer). All standard epi-flourescent filters were used; DAPI, FITC, CY3, Texas Red, and CY5. Whole slide scans were acquired using x10 magnification. The Phenochart software (PerkinElmer) was subsequently used to mark TMA cores from the whole slide scans for subsequent multispectral imaging using x20 magnification. A spectral library was collected by single stainings of colorectal FFPE tissue sections with the CD20 antibody and the individual Opal dyes and subsequent imaging. An unstained sample was used as autofluorescence control and utilized together with the spectral library for spectral unmixing in the inForm software (PerkinElmer). Composite images were compared to single stained slides and inspected for crosstalk and interference.

Edin S., Kaprio T., Hagström J., Larsson P., Mustonen H., Böckelman C., Strigård K., Gunnarsson U., Haglund C, & Palmqvist R. (2019). The Prognostic Importance of CD20+ B lymphocytes in Colorectal Cancer and the Relation to Other Immune Cell subsets. Scientific Reports, 9, 19997.

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