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Vectra multispectral imaging system

Manufactured by PerkinElmer
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The Vectra multispectral imaging system is a high-performance microscopy platform designed for advanced tissue analysis. It features a multispectral imaging capability that enables the simultaneous detection and quantification of multiple biomarkers within a single tissue sample.

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Lab products found in correlation

Inform software, by PerkinElmer (2 mentions) Pa5 28647, by Thermo Fisher Scientific (1 mentions) Vectra 2, by PerkinElmer (1 mentions) Vectashield fluorescence mounting medium, by Vector Laboratories (1 mentions) Spectral dapi, by PerkinElmer (1 mentions) Matlab, by MathWorks (1 mentions) Vectashield antifade mounting medium, by Vector Laboratories (1 mentions) Opal 7 color ihc kit, by Akoya Biosciences (1 mentions) Antifade mounting medium, by Beyotime (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by BD (1 mentions) Bond 3, by Leica (1 mentions) M7103, by Agilent Technologies (1 mentions) Ar9640, by Leica (1 mentions) Inform advanced image analysis software, by PerkinElmer (1 mentions)

Close spelling variants of this product

Vectra Polaris multispectral imaging system Vectra 2.0 multispectral imaging system Vectra 3.0 multispectral imaging system Vectra Automated Multispectral Imaging System Vectra Multispectral Imaging System version 2 Vectra 2 multispectral automated imaging system Vectra Multispectral Imaging System version 3 Multispectral imaging system Vectra imaging system

9 protocols using vectra multispectral imaging system

1

Histological Analysis of Pancreatic Neoplasia

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Formalin-fixed, paraffin-embedded, hematoxylin-and-eosin-stained pancreata of KPC and KPP mice were assessed by a veterinary pathologist employing recommended nomenclature for pancreatic exocrine neoplasia (Hruban et al., 2006 (link)). To better characterize the lesions, pancreas sections from endpoint KPP mice were stained for Alcian blue to detect mucin and glycoproteins, Ki67 (1:200), α-smooth muscle actin (α-SMA, 1:5000), and cytokeratin 19 (CK19, 1:500). The sections were imaged using a Vectra Multispectral Imaging System (Perkin-Elmer).
Talbert E.E., Cuitiño M.C., Ladner K.J., Rajasekerea P.V., Siebert M., Shakya R., Leone G.W., Ostrowski M.C., Paleo B., Weisleder N., Reiser P.J., Webb A., Timmers C.D., Eiferman D.S., Evans D.C., Dillhoff M.E., Schmidt C.R, & Guttridge D.C. (2019). Modeling Human Cancer-induced Cachexia. Cell reports, 28(6), 1612-1622.e4.
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2

Immunohistochemical Analysis of GPER1 in Esophageal Cancer

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Esophageal carcinoma tissue microarrays were purchased from Bioaitech Co., Ltd. (Xi’an, China; ethical license: 2005DKA21300), which contained a total of 148 specimens consisting of EAC (n = 50), ESCC (n = 78), and normal esophagus (NE) tissues (n = 20). The streptavidin–peroxidase method was used for immunohistochemistry analysis of the tissue microarrays. The slides were stained with the GPER1 antibody (PA5-28647; Invitrogen, Carlsbad, CA, USA) [12 (link),52 (link),53 (link)] and visualized with DAB after incubation with the secondary antibody.
The staining results were analyzed using a Vectra Multispectral Imaging System (Perkin Elmer, Waltham, MA, USA). Vectra 2.0.8 (Perkin Elmer) was used for scanning the slides. After importing the high-magnification images and spectral information in inForm 1.2 (Perkin Elmer), positive cells were identified in the target area and the positive rate was determined by setting the threshold intensity. The expression of GPER1 in the tissue microarray is shown with an automatically calculated H-score (=Σpi*i, where i means staining intensity, i = 0,1,2,3; pi is the percentage of the number of cells with corresponding staining intensity in the overall cells). Positive staining was defined as an H-score ≥ 100 (maximum = 300) in this study.
Liu J., Niu Y., Zhang B., Sun Q., Li H., Bai L, & Su Z. (2023). Different Expression Pattern of G Protein-Coupled Estrogen Receptor GPER1 in Esophageal Squamous Cell Carcinoma and Adenocarcinoma. International Journal of Molecular Sciences, 24(18), 14055.
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3

Multiplex Immunofluorescent Staining of TMAs

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Immunofluorescent staining was performed using the opal multiplex immuno-fluorescent system (Perkin Elmer, Hopkinton, MA). Briefly, TMA slides were deparaffinized and tissues were fixed with 10% neutral buffered formalin. Heat- induced epitope retrieval (HIER) was performed in a conventional microwave oven using citrate buffer (pH 6.0). Each section was subjected to six sequential rounds of staining (pan-cytokeratin, DAPI, CD8, CD4, Foxp3, and CD68). Each round of staining included a protein block followed by staining with primary antibody and its corresponding secondary HRP- conjugated polymer. Signal amplification was achieved by using a specific TSA (Tyramide Signal amplification) solution for each antibody. Following this covalent reaction between the labeled tyramide and the tissue, the TMA slides were further subjected to HIER to remove unbound antibodies before the slides were stained with the next antibody. After all five sequential antibody staining, slides were counterstained with spectral DAPI (Perkin Elmer, Hopkinton, MA) and mounted with vectashield fluorescence mounting medium (Vector Labs, Burlingame, CA). Multiplex stained TMA slides were imaged using Vectra multispectral imaging system (Perkin Elmer, Hopkinton, MA) and the images were analyzed using inform software (Perkin Elmer, Hopkinton, MA)
Barua S., Fang P., Sharma A., Fujimoto J., Wistuba I., Rao A.U, & Lin S.H. (2018). Spatial Interaction of Tumor Cells and Regulatory T cells Correlates with Survival in Non-Small Cell Lung Cancer. Lung cancer (Amsterdam, Netherlands), 117, 73-79.
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4

Histological Analysis of Pancreatic Neoplasia

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Formalin-fixed, paraffin-embedded, hematoxylin-and-eosin-stained pancreata of KPC and KPP mice were assessed by a veterinary pathologist employing recommended nomenclature for pancreatic exocrine neoplasia (Hruban et al., 2006 (link)). To better characterize the lesions, pancreas sections from endpoint KPP mice were stained for Alcian blue to detect mucin and glycoproteins, Ki67 (1:200), α-smooth muscle actin (α-SMA, 1:5000), and cytokeratin 19 (CK19, 1:500). The sections were imaged using a Vectra Multispectral Imaging System (Perkin-Elmer).
Talbert E.E., Cuitiño M.C., Ladner K.J., Rajasekerea P.V., Siebert M., Shakya R., Leone G.W., Ostrowski M.C., Paleo B., Weisleder N., Reiser P.J., Webb A., Timmers C.D., Eiferman D.S., Evans D.C., Dillhoff M.E., Schmidt C.R, & Guttridge D.C. (2019). Modeling Human Cancer-induced Cachexia. Cell reports, 28(6), 1612-1622.e4.
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5

Quantifying CD3+ T Cells in CT26 Tumors

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Tumors were fixed in 10% neutral buffered formalin and paraffin-embedded. 5μm tissue sections were deparaffinized, microwaved while boiling for 15 minutes in ph6 citrate antigen retrieval buffer, treated with blocking buffer and incubated with anti-CD3 optimized at a dilution of 1:750 (Abcam, clone SP7). Slides were then incubated with ImmPRESS goat anti-rabbit peroxidase polymer (Vector Laboratories, MP-7451). Signal amplification was performed using Opal 520 tyramide signal amplification reagent (PerkinElmer). A second microwave step was performed to remove labeled primary and secondary antibodies, leaving fluorescent label directly bound to the target site. Slides were counterstained with DAPI and coverslipped using Vectashield antifade mounting medium (Vector Laboratories, H-1400). Slides were scanned at 20× on a Vectra multispectral imaging system (PerkinElmer). Using inForm software (PerkinElmer), the CD3-positive cells and total CT26 cells were quantified, and CD3 cells per 100 tumor cells was determined. Individual 20× fields were stitched into whole slide images using a custom routine in Matlab (Mathworks). All imaging analyses were conducted by personnel who were blinded to the treatment status of the samples.
Qiao G., Qin J., Kunda N., Calata J.F., Mahmud D.L., Gann P., Fu Y.X., Rosenberg S.A., Prabhakar B.S, & Maker A.V. (2017). LIGHT elevation enhances immune eradication of colon cancer metastases. Cancer research, 77(8), 1880-1891.
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6

Multiplex Immunohistochemistry Staining Protocol

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mIHC was performed using an Opal 7‐color IHC kit (Akoya Biosciences, Marlborough, MA, USA) as previously described [21 (link)]. First, the slices were baked, dewaxed, rehydrated, and then subjected to antigen retrieval in AR6 (Perkin Elmer, pH = 6.0) or EDTA (50×, pH = 9.0, Proandy, Xi'an, Shaanxi, China) antigen repair solution in a microwave oven for 10 min and then allowed to cool to room temperature. Then, the sections were blocked in blocking buffer for 10 min, incubated with primary antibody (Supplementary Table S4) for 1 h, and incubated with a polymer HRP‐conjugated secondary antibody for 10 min. Subsequently, the tissues were stained with the fluorophore‐4 tyramine signal amplification dye. After that, the paraffin sections were again subjected to antigen repair. The aforementioned steps were iteratively repeated until all antigens were labeled. Finally, the slides were stained with DAPI (BD Biosciences, San Jose, CA, USA) for 5 min and mounted with antifade mounting medium (Beyotime, Shanghai, China) after elution. Multispectral imaging and segmentation imaging analysis were performed using a Vectra multispectral imaging system (Perkin‐Elmer, Waltham, MA, USA).
Wang J., Zhang J., Liu H., Meng L., Gao X., Zhao Y., Wang C., Gao X., Fan A., Cao T., Fan D., Zhao X, & Lu Y. (2024). N6‐methyladenosine reader hnRNPA2B1 recognizes and stabilizes NEAT1 to confer chemoresistance in gastric cancer. Cancer Communications, 44(4), 469-490.
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7

Multispectral Imaging for Tissue Analysis

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Imaging was performed using a Vectra Multispectral Imaging System (PerkinElmer). One image per core was captured at 200 x magnification. Each 200 x multispectral image cube was created by combining images obtained every 10 nm of the emission light spectrum across the range of each emission filter cube. Five filter cubes were used for each image capture, including DAPI (440–680 nm), FITC (520–680 nm), CY3 (570–690 nm), CY5 (670–720 nm), and Texas Red (580–700 nm).
Pan C., Wang Y., Liu Q., Hu Y., Fu J., Xie X., Zhang S., Xi M, & Wen J. (2021). Phenotypic profiling and prognostic significance of immune infiltrates in esophageal squamous cell carcinoma. Oncoimmunology, 10(1), 1883890.
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8

Dual-Color IHC and Multispectral Imaging for CD8+ T-Cell Analysis

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Two color immunohistochemical staining for human CD8 alpha (Clone C8/144B; Dako M7103; 1:100 dilution) and mesothelin (Clone 5B2, Thermo Scientific MS-1320; 1:30 dilution) was performed sequentially on a Leica Bond III using the Bond Polymer Refine Detection System and the Bond Polymer Refine Red Detection System. Heat-induced epitope retrieval was done for 20 minutes with ER2 solution (Leica Microsystems AR9640). Following dual color immunohistochemistry, multispectral imaging was performed on the stained sections using a Vectra multispectral imaging system (Perkin Elmer, Waltham MA) and the resulting multispectral images were analyzed using InForm Imaging software (Perkin Elmer, Waltham MA). Ten random 20x fields were selected for analysis of each tumor. The images were segmented into tumor and stromal regions using mesothelin staining and morphologic features of the tumor cells from the hematoxylin counterstain using Informs “train by example” segmentation algorithm. CD8+ T-cell counting was then performed by counting CD8+ T cells within tumor and stromal regions.
Wang E., Wang L.C., Tsai C.Y., Bhoj V., Gershenson Z., Moon E., Newick K., Sun J., Lo A., Baradet T., Feldman M.D., Barrett D., Puré E., Albelda S, & Milone M.C. (2015). Generation of Potent T-cell Immunotherapy for Cancer using DAP12-based, Multichain, Chimeric Immunoreceptors. Cancer immunology research, 3(7), 815-826.
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9

Profiling Immune Cells in HPV16+ vHSIL

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Formalin-fixed paraffin-embedded biopsies of 24 HPV16+ vulvar high-grade squamous intraepithelial lesions (vHSIL) patients, treated with an experimental vaccine containing 13 SLPs covering the entire amino acid sequence of HPV16 oncoproteins E6 and E7, were cut in 4 um thick sections. These samples were stained with two multiplex immunofluorescence panels,20 (link) one for T cells and one for myeloid cells. The slides were then scanned with the Vectra multispectral imaging system (PerkinElmer), and cells were, after manual training, automatically phenotyped and counted with inForm advanced image analysis software (PerkinElmer).
Beyranvand Nejad E., Labrie C., Abdulrahman Z., van Elsas M.J., Rademaker E., Kleinovink J.W., van der Sluis T.C., van Duikeren S., Teunisse A.F., Jochemsen A.G., Oosting J., de Miranda N.F., Van Hall T., Arens R, & van der Burg S.H. (2020). Lack of myeloid cell infiltration as an acquired resistance strategy to immunotherapy. Journal for Immunotherapy of Cancer, 8(2), e001326.
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