Evos fl auto fluorescent microscope

Manufactured by Thermo Fisher Scientific

Sourced in United States

The EVOS FL Auto fluorescent microscope is a compact and automated microscope system designed for live-cell imaging and fluorescence microscopy. It features an integrated camera, automated focusing, and a range of objective lenses to enable high-quality imaging of various sample types.

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Lab products found in correlation

Sc 372, by Santa Cruz Biotechnology (1 mentions) Species specific fluorescent secondary antibodies, by Thermo Fisher Scientific (1 mentions) Tubb3, by Thermo Fisher Scientific (1 mentions) Tubb3, by Fortrea (1 mentions) Dapi fluoromount, by Southern Biotech (1 mentions) Mab4470, by R&D Systems (1 mentions)

Close spelling variants of this product

Evos FL Auto 2 fluorescent microscope EVOS FL Auto microscope EVOS FL fluorescent microscope EVOS FL Auto EVOS FL microscope EVOS fluorescent microscope EVOS FL EVOS microscope Fluorescent microscope

7 protocols using evos fl auto fluorescent microscope

Fluorescent Immunostaining of RGCs and AAV-293 Cells

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Primary RGCs and AAV-293 cells were fixed in 4% paraformaldehyde (PFA) for 30 minutes, rinsed in PBS (3 × 10 minutes) and blocked with 5% normal donkey serum with 0.15% Tween-20 in PBS at pH 7.4. Cells were then incubated with either both beta III Tubulin antibody (Tubb3, 1:250, Covance, Denver, PA) and p65 antibody (1:400; sc-372, Santa Cruz Biotechnology, Dallas, TX) or with Tubb3 alone, followed by species-specific secondary fluorescent antibodies (Thermo Fisher Scientific, Grand Island, NY). Negative controls were incubated with secondary antibodies only. The cells were imaged using an EVOS FL Auto fluorescent microscope with onstage incubation capabilities (Thermo Fisher Scientific, Grand Island, NY).

Dvoriantchikova G., Pappas S., Luo X., Ribeiro M., Danek D., Pelaez D., Park K.K, & Ivanov D. (2016). Virally delivered, constitutively active NFκB improves survival of injured retinal ganglion cells. The European journal of neuroscience, 44(11), 2935-2943.

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Potassium-Dependent Dye Uptake Assay

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The low [K⁺] solution (145 mM NaCl, 5 mM KCl, 1.4 mM CaCl₂, 1 mM MgCl₂, 10 mM HEPES diluted in ddH₂O) and high [K⁺] solution (60 mM NaCl, 50 mM KCl, 1.4 mM CaCl₂, 1 mM MgCl₂, 10 mM HEPES diluted in ddH₂O) were prepared as described by [41 (link)]. Cells were washed twice with either the low [K⁺] or high [K⁺] solution, then incubated in fresh low [K⁺] or high [K⁺] solution for 5 min at 37 °C, in 5% CO₂. Next, cells were incubated with 4 mg/ml of sulforhodamine B dye (S1307, Invitrogen, Waltham MA, USA) dissolved in low [K⁺] or high [K⁺] solution for 15 min at 37 °C, in 5% CO₂, washed, and imaged immediately with the EVOS FL Auto fluorescent microscope (Thermo Fisher Scientific) under 10X magnification. Ten random fields of view were imaged per slide. Images were counted for the incidence of dye uptake defined as the percentage of cells that took up the dye.

Freeman E., Langlois S., Leyba M.F., Ammar T., Léger Z., McMillan H.J., Renaud J.M., Jasmin B.J, & Cowan K.N. (2024). Pannexin 1 dysregulation in Duchenne muscular dystrophy and its exacerbation of dystrophic features in mdx mice. Skeletal Muscle, 14, 8.

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Examining PANX1 Role in Myoblast Differentiation

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For the differentiation assays, HSMM (70 000 cells/well) were seeded in 24-well dishes containing one collagen coated glass cover slip. Cells were transduced with a lentivector containing either GFP or PANX1 [46 (link)] one day prior to induction of differentiation. Based on GFP, a transduction efficiency of approximately 50% was achieved. Differentiation was initiated by switching cells into differentiation medium which consisted of DMEM supplemented with 5% horse serum and 1% penicillin/streptomycin. Cells were fixed in 3.7% PFA on day 5 of differentiation. Cells were stained for myosin heavy chain (MHC) (1:500, MF20, MAB4470, R&D Biosystems, Minneapolis, MN, USA), and mounted with Fluoromount-DAPI (Southern Biotech, Birmingham, AL, USA). Cells were imaged with the EVOS FL Auto fluorescent microscope (Thermo Fisher Scientific) under 20X magnification. The differentiation index (number of MHC positive cells/ total number of nuclei (%)) and fusion index (number of nuclei in myotubes/total number of nuclei (%)) were analyzed from 3–5 randomly selected fields of view [34 (link)].

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Fluorescence Microscopy Imaging Protocol

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Detailed fluorescence microscopy methods are described in supplemental methods. Briefly, Fluorescent microscopy images were captured on an EVOS FL auto fluorescent microscope (Thermo Fisher Scientific). GFP (470 ± 22 nm excitation, 510 ± 42 nm emission) and RFP (531 ± 40 nm excitation, 593 ± 40 nm emission) filter cubes were used to capture green or red fluorescence. An EVOS onstage incubator was used for live cell experiments and images were quantitated using ImageJ/Fiji (49 (link),50 (link)) (see supplemental methods, supplemental appendix).

Lant J.T., Kiri R., Duennwald M.L, & O’Donoghue P. (2021). Formation and persistence of polyglutamine aggregates in mistranslating cells. Nucleic Acids Research, 49(20), 11883-11899.

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Visualizing ECM Protein Distribution

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Spatial distribution of ECM proteins, following their covalent attachment to the surface of the PA hydrogels, was visualized following immunolabeling. Briefly, 100 μL of a 0.5 % Triton X-100 (v/v) solution in PBS was added to each well, followed by incubation of the plate at room temperature for 15 min. Gels were washed three times with blank PBS, followed by addition of a 100 μL of 1 % BSA, 0.1 % Tween-20 in PBS. Samples were incubated for 1 hour at room temperature, and then washed three times with PBS. A 100 μL volume of rabbit biotin-labeled anti-fibronectin antibody (1:100) was prepared in labeling buffer (0.5 % BSA, 0.1 % Tween-20 in PBS) and incubated with each well for 1 hour at room temperature. The primary antibody solution was removed and gels washed three times with PBS before addition labeling buffer containing rhodamine-conjugated streptavidin (1:100) to visualize fibronectin coating. After a 1 hour incubation at room temperature the solution was removed and gels washed three times with PBS. Plates were mounted onto an EVOS FL Auto fluorescent microscope (Life Technologies, Grand Island, NY) and imaged at 10x magnification using the manufacturer LED light cube for RFP (531/40 nm excitation, 593/40 nm emission) to visualize fibronectin. Representative images were collected from three independent samples.

Medina S.H., Bush B., Cam M., Sevcik E., DelRio F.W., Nandy K, & Schneider J.P. (2019). Identification of a Mechanogenetic Link between Substrate Stiffness and Chemotherapeutic Response in Breast Cancer. Biomaterials, 202, 1-11.

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Chondrocyte Immunofluorescence Assay

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Chondrocytes were planted at a density of 1 × 10⁴ cells per well in a 24-well plate. In either the presence or absence of 40 µM DHCA, IL-1β (5 ng/ml) was used to treat the cells. For 15 minutes at room temperature, the cells were fixed with 4% paraformaldehyde. Then, cell membranes were punched using a 0.2% Triton X-100 solution for five minutes, and blocked with 5% bovine serum albumin (BSA) for 30 minutes. After that, collagen II, aggrecan, MMP13, and P65-specific antibodies were used to treat the cells overnight at 4°C. After being washed three times with PBS, the cells were incubated with Cy3/Fitc-conjugated secondary antibodies for one hour at 37°C in the dark. The DAPI was left to soak into the cell nuclei for a full ten minutes. The immunofluorescence images were captured using an Evos Fl Auto fluorescent microscope (Life Technologies).

Lu R., Wang Y.G., Qu Y., Wang S.X., Peng C., You H., Zhu W, & Chen A. (2023). Dihydrocaffeic acid improves IL-1β-induced inflammation and cartilage degradation via inhibiting NF-κB and MAPK signalling pathways. Bone & Joint Research, 12(4), 259-273.

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Quantification of 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 to FluoroBrite DMEM. ASC specks were analysed with EVOS FL Auto fluorescent 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 image processing program ImageJ.

Lučiūnaitė A., Dalgedienė I., Žilionis R., Mašalaitė K., Norkienė M., Šinkūnas A., Gedvilaitė A., Kučinskaitė-Kodzė I., & Žvirblienė A. (2021). Activation of NLRP3 Inflammasome by Virus-like Particles of Human Polyomaviruses in Macrophages.

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