Opal 540

Manufactured by Akoya Biosciences

The Opal 540 is a fluorescence-based imaging system designed for multiplexed protein detection and quantification in tissue samples. It enables the simultaneous visualization of multiple protein targets within a single tissue section.

Automatically generated - may contain errors

Lab products found in correlation

2 protocols using opal 540

Multiplex Immunohistochemistry for Tissue Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Slides were stained in accordance with the manufactures protocol (Akoya Biosciences, Catalog #NEL821001KT). The following primary antibodies were used: CD3 (Clone Sp7, Abcam, #ab86734), CD8 (Clone C8/144B, Dako Agilent, Catalog #M710301-2), FoxP3 (Clone 236A/E7, EBioscience, Catalog #14-4777-82), CD20 (Clone L26, ThermoFisher, Catalog #14-0202-82), CD68 (Clone PG-M1, ThermoFisher, Catalog #MA5-12407), CD163 (Clone 10D6, ThermoFisher, Catalog # MA5-11458), Ki-67 (Clone MIB-1, Dako, Catalog #M724029-2), CD14 (Clone EPR3653, Abcam, Catalog #ab133335), PanCK (Clone AE1/AE3 + 5D3, Abcam, Catalog #ab86734), in combination with Opal Polaris 480 (Akoya Biosciences, Catalog #FP1500001KT), Opal Polaris 780 (Akoya Biosciences, Catalog #FP1501001KT) and Opal 520, Opal 540, Opal 570, Opal 620, Opal 650 and Opal 690 (Akoya Biosciences, Catalog #NEL821001KT). Slides were imaged using the Vectra Polaris.

Zwart E.S., van Ee T., Affandi A.J., Boyd L.N., Rodriguez E., den Haan J.M., Farina A., van Grieken N.C., Meijer L.L., van Kooyk Y., Mebius R.E, & Kazemier G. (2023). Spatial immune composition of tumor microenvironment in patients with pancreatic cancer. Cancer Immunology, Immunotherapy : CII, 72(12), 4385-4397.

+ Open protocol

+ Expand

Multiplex Immunofluorescence Tissue Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols

Multiplex immunofluorescence (IF) staining, validation and analysis are detailed in Supplementary Methods and Supplementary Figs. S2 and S3. Briefly, 4-μm sections of formalin-fixed paraffin-embedded (FFPE) tissue samples were deparaffinised and antigen retrieval was performed using DAKO PT-Link heat-induced antigen retrieval with a low- (pH 6) or high-pH (pH 9) solution (DAKO). Samples were stained with mAbs targeting cytokeratin (CK), CD4, CD8, FOXP3, PD-1 and CD137 followed by TSA visualisation with fluorophores Opal 520, Opal 540, Opal 570, Opal 620, Opal 650 and Opal 690 (Akoya Biosciences), as described earlier.²⁰ Each tissue section was put through several sequential rounds of antibody staining. In the seventh round, nuclei were counterstained with spectral DAPI (Akoya Biosciences) and sections mounted with Faramount Aqueous Mounting Medium (Dako). Multiplexed immunofluorescence slides were scanned on a Vectra-Polaris Automated Quantitative Pathology Imaging System (Akoya Biosciences). Tissue imaging, spectral unmixing and phenotyping were performed using inForm software (version 2.4.8, Akoya Biosciences), as described previously.²⁰

Salas-Benito D., Conde E., Tamayo-Uria I., Mancheño U., Elizalde E., Garcia-Ros D., Aramendia J.M., Muruzabal J.C., Alcaide J., Guillen-Grima F., Minguez J.A., Amores-Tirado J., Gonzalez-Martin A., Sarobe P., Lasarte J.J., Ponz-Sarvise M., De Andrea C.E, & Hervas-Stubbs S. (2021). The mutational load and a T-cell inflamed tumour phenotype identify ovarian cancer patients rendering tumour-reactive T cells from PD-1+ tumour-infiltrating lymphocytes. British Journal of Cancer, 124(6), 1138-1149.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!