Efluor 780 fixable dye

Manufactured by Thermo Fisher Scientific

The EFluor™ 780 fixable dye is a fluorescent dye used for labeling and detecting cells in flow cytometry applications. It provides a stable, covalent label that remains bound to the cells even after fixation and permeabilization. The dye has an excitation maximum at 780 nm and an emission maximum at 810 nm, making it suitable for detection in the far-red region of the spectrum.

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Lab products found in correlation

Canto flow cytometer, by BD (1 mentions) Facs diva software, by FlowJo (1 mentions)

Close spelling variants of this product

Fixable Viability Dyes eFluor 780 EBioscience Fixable Viability Dyes eFluor™ 506 and 780 EFluor 780

2 protocols using efluor 780 fixable dye

Liver Immune Cell Isolation

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To analyze early changes in the TIME, mice were injected with 5 × 10⁵ tumor cells via the intrasplenic/portal route, and the livers removed 3, 7, or 9 days later (as indicated). Liver homogenates were prepared in cold PBS and filtered through a stainless steel mesh using a plunger. The filtrates were centrifuged at 60 G to separate the hepatocytes, the supernatants containing the nonparenchymal cell fraction centrifuged at 480 g and the pellets resuspended in 10 ml of a 37.5% Percoll solution in HBSS containing 100 Uml⁻¹ heparin and centrifuged at 850 g for 30 min to obtain the immune cell-rich fraction. Prior to FC, red blood cells were removed using the ACK (ammonium–chloride–potassium) solution and 1 × 10⁶ cells were immunostained with the indicated antibodies. Data acquisition was with a BD Canto flow cytometer and FACS Diva software and the data analyzed using the FlowJo software. For flow cytometric experiments on hepatic leukocytes, single cells were gated based on size (forward scatter), granularity (side scatter) and viability using an eFluor™ 780 fixable dye (eBioscience™, ThermoFisher).

Milette S., Hashimoto M., Perrino S., Qi S., Chen M., Ham B., Wang N., Istomine R., Lowy A.M., Piccirillo C.A, & Brodt P. (2019). Sexual dimorphism and the role of estrogen in the immune microenvironment of liver metastases. Nature Communications, 10, 5745.

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Isolation and Analysis of Hepatic Immune Cells

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To analyze early changes in the TIME, mice were injected with 1x10⁵ tumor cells via the intrasplenic/portal route and the livers removed 14 days later (or as indicated). Liver homogenates were prepared in cold PBS and filtered through a stainless steel mesh, using a plunger. The filtrates were centrifuged at 500 rpm to separate the hepatocytes, the supernatants containing the non-parenchymal cell fraction centrifuged at 1400 rpm and the pellets resuspended in 10 ml of a 37.5% Percoll solution in HBSS containing 100 U/ml heparin and centrifuged at 1910 rpm for 30 minutes to obtain the immune cell–rich fraction. Prior to flow cytometry, red blood cells were removed using the ACK (ammonium chloride-potassium) solution and 1x10⁶ cells were immunostained with the indicated antibodies. Data acquisition was with a BD LSRFortessa and FACS Diva software and the data analyzed using the FlowJo software. For FC on hepatic leukocytes, single cells were gated based on size (FSC), granularity (SSC), viability using an eFluor™ 780 fixable dye (eBioscience™, Thermofisher) and the expression of CD45.

Hashimoto M., Konda J.D., Perrino S., Fernandez M.C., Lowy A.M, & Brodt P. (2021). Targeting the IGF-axis potentiates immunotherapy for pancreatic ductal adenocarcinoma liver metastases by altering the immunosuppressive microenvironment. Molecular cancer therapeutics, 20(12), 2469-2482.

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