Evos fluorescent microscope

Manufactured by Thermo Fisher Scientific

Sourced in United States

The EVOS fluorescent microscope is a high-performance imaging system designed for advanced fluorescence applications. It features an LED light source, digital image capture, and intuitive software for visualization and analysis of fluorescently labeled samples.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc antibody, by BioLegend (1 mentions) Anti phospho histone h3, by New England Biolabs (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Tnf α, by R&D Systems (1 mentions) Importazole, by Merck Group (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Recombinant mouse il 17a, by R&D Systems (1 mentions) Il 17a, by R&D Systems (1 mentions) Donkey anti rabbit alexa 488, by Thermo Fisher Scientific (1 mentions) Rabbit anti ki67, by Abcam (1 mentions) Nunc lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions) Ifn gamma, by Cell Signaling Technology (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Ab125212, by Abcam (1 mentions) Ab178847, by Abcam (1 mentions) Ab104225, by Abcam (1 mentions) Ab6326, by Abcam (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions) Alexa fluor 594 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)

21 protocols using evos fluorescent microscope

Proliferation, Mitosis, and Apoptosis Assays

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For proliferation assays, cells were plated in a 96-well plate, 1000 cells/well, with 8 replicates of each sample. The sulforhodamine-B proliferation assay was used.
Mitotic index was measured by fixation of cells and staining with DAPI (Fisher Scientific) and anti-phospho-Histone-H3 (NEB, Hitchin, UK).
Chromosome counts were performed by trypsinizing the cells, treating them with a hypotonic solution, and then spreading them onto a microscope slide. The DNA was then stained with DAPI, and imaged using an EVOS Fluorescent Microscope (Fisher Scientific). Images were analyzed using the FIJI release of ImageJ (ver. 1.50a) (46 (link)). Apoptosis was measured using an Annexin-V-FITC antibody (Biolegend UK) and a CYAN ADP flow cytometer using standard methods.

Jones P.D., Kaiser M.A., Ghaderi Najafabadi M., McVey D.G., Beveridge A.J., Schofield C.L., Samani N.J, & Webb T.R. (2016). The Coronary Artery Disease-associated Coding Variant in Zinc Finger C3HC-type Containing 1 (ZC3HC1) Affects Cell Cycle Regulation. The Journal of Biological Chemistry, 291(31), 16318-16327.

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Quantifying THP-1 Monocyte Adhesion

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THP-1 monocytes were cultured in RPMI-1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin. Cells were pelleted, resuspended to a concentration of 1 x 10⁵ cells/ml in full medium. Cells were labelled with 5mM Calcein-AM (Fisher Scientific, C3099) for 30 minutes. The HUVEC monolayers were treated with TNFα (R&D Systems, 210-TA-005) for 30 minutes as appropriate, washed with RPMI-1640, then 1x10⁴ THP-1 cells added and incubated for 30 minutes. The THP-1s were then removed and the THP-1-bound monolayer washed 4 times with RPMI-1640. The final wash was removed, then the cells fixed with 4% paraformaldehyde. Images were taken with an EVOS Fluorescent Microscope (Fisher Scientific) and the number of bound cells counted manually in ImageJ.

Jones P.D., Kaiser M.A., Ghaderi Najafabadi M., Koplev S., Zhao Y., Douglas G., Kyriakou T., Andrews S., Rajmohan R., Watkins H., Channon K.M., Ye S., Yang X., Björkegren J.L., Samani N.J, & Webb T.R. (2018). The JCAD gene at the 10p11 coronary artery disease locus regulates Hippo signaling in endothelial cells. Arteriosclerosis, thrombosis, and vascular biology, 38(8), 1711-1722.

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Overexpression of TRIP13 in IMCD-3 Cells

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Full length Trip13 cDNA were cloned into pEGFP‐C1 using standard cloning techniques, and transfected into IMCD‐3 cells using Lipofectamine 3000 (Invitrogen Life Technologies, Carlsbad, CA). A mixed population of genetically modified cells were obtained following incubation with DMEM:F12 containing Geneticin (500–750 μg/mL). The genetically modified IMCD‐3 cells expressing full‐length TRIP13 (1–432) were examined for EGFP fluorescence using EVOS fluorescent microscope (Life Technologies) at 10–40X magnification. In some experiments, the genetically modified IMCD‐3 cells were studied in the presence and absence of H₂O₂ (8.8–88 μM) and importazole (50–75 μM; Sigma‐Aldrich, St. Louis, MO).

Pressly J.D., Hama T., Brien S.O., Regner K.R, & Park F. (2017). TRIP13-deficient tubular epithelial cells are susceptible to apoptosis following acute kidney injury. Scientific Reports, 7, 43196.

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Quantifying OPC Proliferation with IL-17A

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OPCs were plated on poly-D-lysine-coated eight-well glass chamber slides (BD Biosciences). After 48 h half of the OPC medium was replaced with or without 2x recombinant mouse IL-17A for a final concentration of 0.01, 0.1, 1, 10, or 100 ng/mL IL-17A (R&D Systems). After an additional 48 h, cells were washed with PBS and fixed with 0.5% paraformaldehyde for 30 min at RT. For immunocytochemistry cells were blocked with 10% donkey serum/1% BSA then stained for Ki67 (1:500 rabbit-anti-Ki67, Abcam; 1:1000 Alexa 488 donkey-anti-rabbit, Life Technologies). Cells were counter-stained with DAPI and imaged using an EVOS fluorescent microscope (Life Technologies) with 20× and 40× magnification objectives. Ki67⁺ cells were counted out of 200 cells in each well. The scorer was blinded to the experimental group. Proliferation was calculated as the percentage of Ki67⁺ cells out of total cells.

Rodgers J.M., Robinson A.P., Rosler E.S., Lariosa-Willingham K., Persons R.E., Dugas J.C, & Miller S.D. (2014). IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells. Glia, 63(5), 768-779.

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Enumeration of Microbial Cells in Aquatic Environments

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Cell density was determined in both the pool and vent waters collected in 2020 using fluorescent microscopy. Ten milliliters of water was subsampled in triplicate with a sterile syringe and transferred to sterile 15 mL tubes containing formaldehyde to a final concentration of 2% v/v. At the laboratory, 1.5 µL of SYBR Gold (100× stock solution) and 2 µL of DAPI (2 mg mL⁻¹) were added to each 10 mL water sample and these were incubated in the dark at room temperature for 20 min. Stained cells were collected on 0.22 µm black polycarbonate filters (Millipore, Billerica, MA, USA) and were visualized using an EVOS fluorescent microscope (Life Technologies, Carlsbad, CA, USA). DAPI- and SYBR Gold–stained cells were imaged, and images of filtered cells were overlayed using the “overlay” function in the EVOS software package to better differentiate debris and minerals in the water from cells. Thirty images were randomly captured, and cell counts for each water type (pool and vent) were performed in triplicate.

Keller L.M., Colman D.R, & Boyd E.S. (2023). An active microbiome in Old Faithful geyser. PNAS Nexus, 2(3), pgad066.

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Immunofluorescence Assay for E-cadherin and PD-L1

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Cells were seeded in an 8-well Nunc Lab-Tek II Chamber Slide System (Thermo Scientific, Rochester, NY), and treated the day after with PBS or IFN-gamma (Cell Signaling Technology) for 24 hours. Immunofluorescence was performed according to the manufacture’s protocol. Briefly, cells were fixed with 4% formaldehyde, blocked with blocking buffer, and incubated with primary antibodies for E-cadherin (mouse monoclonal) or PD-L1 (rabbit monoclonal) at 4°C overnight. After the washing with PBS, cells were incubated with fluorochrome-conjugated secondary antibodies for anti-rabbit and anti-mouse (Cell Signaling Technology). Cell nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Microscopy was performed on an EVOS fluorescent microscope (Model FL, Life Technologies, Carlsbad, CA).

Suda K., Rozeboom L., Rivard C.J., Yu H., Ellison K., Melnick M.A., Hinz T.K., Chan D., Heasley L.E., Politi K., Mitsudomi T, & Hirsch F.R. (2017). Therapy-induced E-cadherin downregulation alters expression of programmed death ligand-1 in lung cancer cells. Lung cancer (Amsterdam, Netherlands), 109, 1-8.

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Immunofluorescence Staining of Teratoma

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Teratoma sections were de-paraffinized, fixed in 4% paraformaldehyde, and stained with antibodies targeted against alpha fetoprotein (R & D Systems, Minneapolis, MN; 1:100), glial fibrillary acidic protein (GFAP; Life Technologies; 1:100), and smooth muscle actin (Abcam; 1:100). Slides were coverslipped as described above and imaged on an EVOS fluorescent microscope (Life Technologies).

Burnight E.R., Wiley L.A., Drack A.V., Braun T.A., Anfinson K.R., Kaalberg E.E., Halder J.A., Affatigato L.M., Mullins R.F., Stone E.M, & Tucker B.A. (2014). CEP290 gene transfer rescues Leber Congenital Amaurosis cellular phenotype. Gene therapy, 21(7), 662-672.

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Transfection Efficiency Evaluation

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All the cell lines used in this study were assessed for transfection efficiency using GFP expressing plasmid pGLO (Bio-Rad, Hercules, CA, USA). Each cell line was seeded (3 × 10⁴) in 48-well plates and transfected with 100 ng/well pGLO plasmid using Lipofectamine 2000 (Invitrogen, Grand Island, NY, USA) according to manufacturer’s instructions. After 24 h post-transfection, cells were visualized and images were captured on an EVOS fluorescent microscope (Life Technologies, Grand Island, NY, USA). GFP-positive cells were determined using flow cytometry and the percent positive population was analyzed using FlowJo software (Ashland, OR, USA).

Kulkarni V., Naqvi A.R., Uttamani J.R, & Nares S. (2016). MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines. International Journal of Molecular Sciences, 17(1), 72.

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Immunofluorescence Staining of Neural Markers

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Immunofluorescence staining was conducted as previously described (13 (link)). Staining was conducted overnight at 4°C with anti-NeuN (1:500; ab104225; Abcam, Waltham, MA), rabbit anti-CD68 (1:500; ab125212; Abcam), rat anti-BrdU (1:500; ab6326; Abcam) and/or rabbit anti-Iba1 (1:500; ab178847; Abcam) primary antibodies. Sections were then incubated with appropriate secondary antibodies for 60 mins at room temperature (1:500; Alexa Fluor goat anti-rabbit 594, A11037; Alexa Fluor goat anti-rabbit 488, A11008; Alexa Fluor goat anti-rat 594, A11007; Invitrogen, Waltham, MA) before mounting with DAPI Fluoromount-G (0100–20; SouthernBiotech, Birmingham, AL). Fluorescent images were captured with either an EVOS fluorescent microscope (ThermoFisher Scientific, Waltham, MA) or BZ-X800E fluorescent microscope (Keyence, Itasca, IL). Image analysis was performed using FIJI (ImageJ/Fiji, version 1.53o; National Institutes of Health, Bethesda, MD).

Bosco D.B., Kremen V., Haruwaka K., Zhao S., Wang L., Ebner B.A., Zheng J., Dheer A., Perry J.F., Xie M., Nguyen A.T., Worrell G.A, & Wu L.J. (2024). Impaired microglial phagocytosis promotes seizure development. bioRxiv.

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SARS-CoV-2 RBD Immunofluorescence Assay

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After the first and third passage, Vero E6 monolayers were fixed and stained with mouse monoclonal antibodies specific for the SARS-CoV-2 RBD region and a fluorescein-labeled goat anti-mouse antibody as previously described [11 ]. Monolayers were examined for fluorescein isothiocyanate-positive cells with an EVOS fluorescent microscope (Thermo Fischer Scientific, Waltham, MA, USA). Mock-infected and SARS-CoV-2 infected Vero E6 cells were used as negative and positive controls, respectively, as shown in Additional file 1: Figure S1.

Balaraman V., Drolet B.S., Mitzel D.N., Wilson W.C., Owens J., Gaudreault N.N., Meekins D.A., Bold D., Trujillo J.D., Noronha L.E., Richt J.A, & Nayduch D. (2021). Mechanical transmission of SARS-CoV-2 by house flies. Parasites & Vectors, 14, 214.

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