Evos fl auto fluorescence microscope

Manufactured by Thermo Fisher Scientific

Sourced in United States

The EVOS FL Auto fluorescence microscope is a versatile imaging system designed for live-cell and fixed-sample analysis. It provides high-quality fluorescence imaging capabilities with a range of objectives and filter sets. The microscope is equipped with automated functions for enhanced imaging performance and ease of use.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Mb49 murine bladder carcinoma cell line, by Merck Group (2 mentions) Umuc 3 and t24 human urinary bladder cancer cell lines, by American Type Culture Collection (2 mentions) Renca murine kidney adenocarcinoma cell line, by American Type Culture Collection (2 mentions) Mycoplasma test, by Lonza (2 mentions) Glowcell 16 fluc f2a gfp lentivirus, by Biosettia (2 mentions) Triton x 100, by Merck Group (2 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (1 mentions) A0564, by Agilent Technologies (1 mentions) Tunel system, by Promega (1 mentions) Gw2580, by Abcam (1 mentions) Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions) Alkaline phosphatase detection kit, by Merck Group (1 mentions) 6 diamidino 2 phenylindole dapi, by Merck Group (1 mentions) Nucblue live readyprobe reagent, by Thermo Fisher Scientific (1 mentions) Costar transwell permeable support, by Corning (1 mentions) Matrigel, by R&D Systems (1 mentions) Hoechst 33342, by Abcam (1 mentions)

Close spelling variants of this product

EVOS FL Auto (301 citations) EVOS FL (286 citations) EVOS FL microscope (227 citations) EVOS microscope (218 citations) EVOS FL Auto microscope (152 citations) Fluorescence microscope (108 citations) EVOS fluorescence microscope (97 citations) EVOS FL fluorescence microscope (57 citations) EVOS FL Auto Fluorescence Inverted Microscope (5 citations) Evos FL Auto 2 fluorescence microscope (3 citations)

35 protocols using evos fl auto fluorescence microscope

Immunofluorescence Assay for ABCB1 and ABCG2

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The immunofluorescence assay was performed as we previously described [32 ]. After cultured overnight in 24-well plates, the cells (2 × 10⁴) were treated with selonsertib for 0 h, 24 h, 48 h, and 72 h respectively. Then cells were fixed in 4% paraformaldehyde for 10 min and permeabilized by 0.1% Triton X-100 for 10 min before blocked with 6% BSA for 1 h. The presence of ABCB1 and ABCG2 was determined using monoclonal antibody F4 (dilution 1:100) and BXP-21 (dilution 1:150) respectively for incubation at 4 °C overnight. Alexa Fluor 488 conjugated secondary antibody (1:1000) was used after washing with iced PBS, DAPI was used to counterstain the nuclei. Immunofluorescence images were collected using an EVOS FL Auto fluorescence microscope (Life Technologies Corporation, Gaithersburg, MD).

Ji N., Yang Y., Cai C.Y., Lei Z.N., Wang J.Q., Gupta P., Shukla S., Ambudkar S.V., Kong D, & Chen Z.S. (2018). Selonsertib (GS-4997), an ASK1 inhibitor, antagonizes multidrug resistance in ABCB1- and ABCG2-overexpressing cancer cells. Cancer letters, 440-441, 82-93.

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Alkaline Comet Assay for Genotoxicity

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Alkaline Comet assay was performed using Trevigen’s Comet assay kit (#4250–050-K). Briefly, Dox-induced cells and empty vector cells treated with and without metals were lysed, resuspended in low melting agarose, and spread evenly onto Comet slides. The slides were immersed in freshly prepared alkaline unwinding solution prior to alkaline electrophoresis. DNA in the nucleoid was visualized with SYBR Gold staining using an EVOS FL auto-fluorescence microscope (Life Technologies). Comet tail moment was scored using the Open Comet plugin for ImageJ software with 50 randomly selected cells.

Vasquez V., Mitra J., Hegde P.M., Pandey A., Sengupta S., Mitra S., Rao K.S, & Hegde M.L. (2017). Chromatin-Bound Oxidized α-Synuclein Causes Strand Breaks in Neuronal Genomes in in vitro Models of Parkinson’s Disease. Journal of Alzheimer's disease : JAD, 60(Suppl 1), S133-S150.

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Apoptosis Evaluation in HeLa Cells

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The Live Cell Caspase-3 Activity and Annexin V Apoptosis Detection Kit (Beyotime, Shanghai, China) were used to evaluate HeLa cell apoptosis. Briefly, HeLa cells were cultured in a 12-well plate for 24 h at 37 °C and then treated with 20(S)-GRh2 (45 μM) for 24 h. HeLa cells were then incubated with Caspase-3 substrate (5 μM) and Annexin V–mCherry (5 μM) for 30 min in the dark at room temperature. The cells were washed twice with PBS and analyzed immediately under an EVOS FL autofluorescence microscope (Life Technologies, Carlsbad, CA, USA).

Liu Y., Wang X., Qiao J., Wang J., Jiang L., Wang C., Yu S., Zhang P., Zhao D., Fan M, & Liu M. (2022). Ginsenoside Rh2 Induces HeLa Apoptosis through Upregulating Endoplasmic Reticulum Stress-Related and Downstream Apoptotic Gene Expression. Molecules, 27(22), 7865.

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Immunofluorescence Analysis of ABCB1 and ABCG2 Expression

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The immunofluorescence assay was performed as previously described (Zhang X. Y. et al., 2016 (link)). After being cultured overnight in 24-well plates, the cells (2 × 10⁴) were treated with VS-4718 for 0, 24, 48, and 72 h. Then cells were fixed in 4% paraformaldehyde for 10 min and permeabilized by 0.1% Triton X-100 for 10 min before blocking with 6% BSA for 1 h. Cells were incubated with monoclonal antibodies ABCB1 (F4, dilution 1:100) and ABCG2 (BXP-21, dilution 1:150) at 4 °C overnight. Alexa Fluor 488 conjugated secondary antibody (1:1,000) was used after washing with iced PBS. DAPI was used to counterstain the nuclei. Immunofluorescence images were collected using an EVOS FL Auto fluorescence microscope (Life Technologies Corporation, Gaithersburg, MD).

Ji N., Yang Y., Cai C.Y., Lei Z.N., Wang J.Q., Gupta P., Teng Q.X., Chen Z.S., Kong D, & Yang D.H. (2018). VS-4718 Antagonizes Multidrug Resistance in ABCB1- and ABCG2-Overexpressing Cancer Cells by Inhibiting the Efflux Function of ABC Transporters. Frontiers in Pharmacology, 9, 1236.

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Breast Cancer Cell Imaging Assay

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MDA-MB-468 or MDA-MB-231 cells (5 × 10³ cells/well) were seeded on 8-well Ibidi chamber slides overnight and then treated with PBS (control), Sal (0.5 μM), Das (15 μM), or the drug combination (0.5 + 15 μM) for different time intervals (6, 12, 24, 48, and 72 h). DAPI (9 μM), DCF-DA (10 μM), or phycoerythrin-labeled anti-estrogen receptor β antibody (1:100 dilution) were used for staining the nucleus, the iROS, or the estrogen receptors β (ESR2), respectively, 30 min before imaging. The cells were then washed with PBS. The fluorescence images were acquired using EVOS FL Auto Fluorescence Microscope (Life Technologies, Carlsbad, CA).

Bellat V., Verchère A., Ashe S.A, & Law B. (2020). Transcriptomic insight into salinomycin mechanisms in breast cancer cell lines: synergistic effects with dasatinib and induction of estrogen receptor β. BMC Cancer, 20, 661.

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Immunofluorescence Staining of 2D Cell Cultures

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For 2D culture, cells were plated in 8-well chamber slides and kept in conditioned medium [13 (link)] (three volumes of feeder cell-conditioned F-medium mixed with one volume of fresh F-medium supplemented with 5 μM Y-27632) to grow in a monolayer and then fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.5% Triton X-100 for 15 minutes, and blocked with 5% bovine serum albumin for one hour at room temperature. After blocking, cells were incubated overnight at 4°C with primary antibodies (Supplementary Table 2). Alexa Fluor (AF) 594-(red, Molecular Probes, Eugene, OR) or AF488-conjugated secondary antibody (green, Molecular Probes) were used to visualize the signal. Following three washes with PBS, slides were mounted with the VECTASHIELD mounting medium with DAPI (Vector Laboratories, Burlingame, CA). The fluorescence images were taken using the EVOS FL Auto fluorescence microscope (Life Technologies, Carlsbad, CA).

Jin L., Qu Y., Gomez L.J., Chung S., Han B., Gao B., Yue Y., Gong Y., Liu X., Amersi F., Dang C., Giuliano A.E, & Cui X. (2017). Characterization of primary human mammary epithelial cells isolated and propagated by conditional reprogrammed cell culture. Oncotarget, 9(14), 11503-11514.

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Quantifying Islet Cell Apoptosis

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Paraffin-embedded islet grafts within the kidneys or intact mouse or human islets were stained for insulin using polyclonal guinea pig anti-insulin antibody (Dako, A0564), and apoptosis was measured using the TUNEL system (Promega, #G3250). Images from each islet graft were obtained using an EVOS FL Autofluorescence microscope (Life Technologies). ImageJ software was used to quantify total insulin-positive cells and TUNEL-insulin co-staining cells. Image quantification was done blinded to experimental group. A total of 7,471 β-cells were counted from control mouse grafts and 8,837 β-cells were counted from CCK treated mouse grafts.

Kim H.T., de Souza A.H., Umhoefer H., Han J., Anzia L., Sacotte S.J., Williams R.A., Blumer J.T., Bartosiak J.T., Fontaine D.A., Baan M., Kibbe C.R, & Davis D.B. (2021). Cholecystokinin Attenuates β-Cell Apoptosis in both Mouse and Human Islets. Translational research : the journal of laboratory and clinical medicine, 243, 1-13.

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Quantifying P-gp Expression in Cancer Cells

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SW620, SW620/Ad300 cells were seeded (1 × 10⁴/well) in 24-well plates and cultured at 37°C for 24 h, followed by incubation with 3 μM glesatinib for 0, 24, 48, and 72 h, respectively. Then cells were fixed in 4% paraformaldehyde for 5 min and permeabilized by 0.1% Triton X-100 for 5 min before blocked with 6% BSA for 1 h at 37°C. The presence of P-gp was determined using monoclonal antibody F4 (dilution 1:1000) for incubation at 4°C overnight. Alexa Fluor 488 conjugated secondary antibody (1:1000) was used for incubation at 37°C for 1 h. After washing with iced PBS, DAPI (1 μg/mL) was used to counterstain the nuclei. Immunofluorescence images were collected using an EVOS FL Auto fluorescence microscope (Life Technologies Corporation, Gaithersburg, MD).

Cui Q., Cai C.Y., Gao H.L., Ren L., Ji N., Gupta P., Yang Y., Shukla S., Ambudkar S.V., Yang D.H, & Chen Z.S. (2019). Glesatinib, a c-MET/SMO Dual Inhibitor, Antagonizes P-glycoprotein Mediated Multidrug Resistance in Cancer Cells. Frontiers in Oncology, 9, 313.

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Transduction of Bladder and Kidney Cancer Cell Lines

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MB49 murine bladder carcinoma cell line (Cat# SCC148) was supplied by EMD Millipore Corporation (Temecula, CA). UMUC-3 and T24 human urinary bladder cancer cell lines (Cat# CRL-1749 and Cat# HTB-4), and Renca murine kidney adenocarcinoma cell line (Cat# CRL-2947), were obtained from ATCC (Manassas, VA). Each cell line was cultured and maintained (no more than 10 passages after initial thawing) according to the company’s instructions. Mycoplasma tests (Lonza, Basel, Switzerland) were performed periodically to ensure that there was no contamination. Both UMUC-3 and Renca cell lines were further transduced with GlowCell 16 FLuc-F2A-GFP lentivirus (Biosettia, San Diego, CA) carrying both firefly luciferase and green fluorescent protein (GFP) genes. Briefly, the cells were seeded in 6-well plates (0.25 × 10⁶ cells/well) and incubated for 3 days with the virus (2×10⁷ IU/well) in the presence of polybrene (10 μg/mL). To ensure more than 95% of cell purity, the cell lines were analyzed and sorted for GFP expression using flow cytometry. The successful transduction was further confirmed by imaging, using an EVOS FL Auto Fluorescence microscope (Life Technology, Carlsbad, CA).

Bellat V., Michel A.O., Thomas C., Stokol T., Choi B.B, & Law B. (2022). A Urinary Drug-Disposing Approach as an Alternative to Intravesical Chemotherapy for Treating Nonmuscle Invasive Bladder Cancer. Cancer Research, 82(7), 1409-1422.

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Quantifying ASC Speck Formation

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THP-1 monocytes expressing ASC fused with green fluorescent protein (GFP) was used for ASC speck formation. The cells were cultivated in RPMI 1640 medium supplemented with 10% FBS, 1% P/S, 100 μg/ml normocin, and 100 μg/ml selective antibiotic zeocin. THP-1-ASC-GFP cells were differentiated to macrophages as origin THP-1 using RPMI 1640 medium supplemented with 10% FBS, 1% P/S, and PMA. THP-1-ASC-GFP macrophages were treated with viral proteins in serum-free RPMI. Thirty minutes before the treatment termination, Hoechst33342 was added to stain cell nuclei. After 24 h of treatment, cell culture medium was replaced with FluoroBrite DMEM. ASC specks were analyzed with an EVOS FL Auto fluorescence microscope (Life Technologies, USA) by taking photos with 20× objective. Cells were counted according to the number of nuclei. ASC speck number per cell was counted using the image processing program ImageJ.

Lučiūnaitė A., Dalgėdienė I., Žilionis R., Mašalaitė K., Norkienė M., Šinkūnas A., Gedvilaitė A., Kučinskaitė-Kodzė I, & Žvirblienė A. (2022). Activation of NLRP3 Inflammasome by Virus-Like Particles of Human Polyomaviruses in Macrophages. Frontiers in Immunology, 13, 831815.

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