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Evos m500

Manufactured by Thermo Fisher Scientific
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The EVOS M500 is a digital inverted microscope designed for cell and tissue imaging. It features a high-resolution CMOS camera and LED illumination for capturing clear, detailed images. The microscope supports brightfield, phase contrast, and fluorescence imaging modes to accommodate a variety of sample types.

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Lab products found in correlation

Clear round bottom ultra low attachment microplate, by Corning (2 mentions) Filtermax f5, by Molecular Devices (1 mentions)

10 protocols using evos m500

1

Assessing BMA Potency Against Trypanosoma Cruzi

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For the
IC50 assay of BMA, trypomastigotes were added
at 1 × 106 cells/well in 96-well microplates and treated
(maximum concentration of 150 μM) for 24 h. Resazurin (0.011%
in phosphate-buffered saline (PBS)) was used to detect parasite viability.23 (link) Untreated trypomastigotes were used as 100%
viability. The analysis was performed at 570 nm using a spectrophotometer
(FilterMax F5, Molecular Devices). Benznidazole was applied as the
control drug.
The IC50 assay in the amastigote forms
was obtained using peritoneal macrophages in 16-well plates (Thermo)
at 1 × 105 cells/well. Macrophages were infected with
trypomastigotes (1:10 ratio) for 2 h23 (link) and
treated with BMA (2.03–65.0 μM) for 48 h.
Giemsa was used for staining and the material was analyzed with a
digital microscope (EVOS M500, Thermo). The infection index was obtained
in 200 macrophages. Benznidazole was used as the standard.57 (link)
Romanelli M.M., Amaral M., Thevenard F., Santa Cruz L.M., Regasini L.O., Migotto A.E., Lago J.H, & Tempone A.G. (2022). Mitochondrial Imbalance of Trypanosoma cruzi Induced by the Marine Alkaloid 6-Bromo-2′-de-N-Methylaplysinopsin. ACS Omega, 7(32), 28561-28570.
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2

Quantifying Lung Metastasis Progression

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Tumor monitoring and measurements were performed as described in Velazquez et al. [25 (link)]. The area of the lung metastases were calculated through microscopy (EVOS M500, Thermo Fisher Scientific, Waltham, MA, USA).
Velazquez F.N., Viscardi V., Montemage J., Zhang L., Trocchia C., Delamont M.M., Ahmad R., Hannun Y.A., Obeid L.M, & Snider A.J. (2021). A Milk-Fat Based Diet Increases Metastasis in the MMTV-PyMT Mouse Model of Breast Cancer. Nutrients, 13(7), 2431.
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3

Mitochondrial ROS Quantification in ESCs

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Mitochondrial ROS was detected by the MitoSOX Red probe (MedChemExpress, HY-D1055) according to the manufacturer’s instructions. ESCs cultured in 6-well dishes were stained in situ with 1 µM MitoSOX Red staining solution for 30 min at 37 °C. After washing three times with serum-free medium, the stained cells from each group (n = 5) were observed using a fluorescent microscope (EVOS M500, Thermo Scientific) at an excitation/emission wavelength of 510/580 nm. 5 random fields of cells in each well were evaluated, and the mean fluorescence intensity was analyzed by Image-Pro Plus 6.0 software.
Chen P., Ye C., Huang Y., Xu B., Wu T., Dong Y., Jin Y., Zhao L., Hu C., Mao J, & Wu R. (2024). Glutaminolysis regulates endometrial fibrosis in intrauterine adhesion via modulating mitochondrial function. Biological Research, 57, 13.
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4

Quantitative Analysis of Tumor Spheroid Growth

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2.5 × 103 HeLa cells were seeded in 96-well Clear Round Bottom Ultra-Low Attachment Microplates (7007, Corning®) in a final volume of 200 μl. Cells were centrifuged at 2,000 rpm for 10 min to encourage the formation of a single spheroid per well. 12 replicates of each condition were plated in each biological replicate. Every three days, 100 μl cell culture medium was replaced, and every two days spheroids were photographed in an EVOS™ M500 (Thermo Fisher Scientific™) at 4× magnification. Spheroids were cultured for 7 days. Fiji Image J Software was used to measure the volume and circularity of spheroids. Spheroid area was calculated as previously reported61 (link), and used to determine spheroid radius (R = √(area/π)), from which the volume (V = (4/3) πR3) was calculated. Each spheroid’s GFP levels were quantified using Fiji Image J Software and the intensity relative to spheroid volume was depicted.
Ortmann B.M., Burrows N., Lobb I.T., Arnaiz E., Wit N., Bailey P.S., Jordon L.H., Lombardi O., Peñalver A., McCaffrey J., Seear R., Mole D.R., Ratcliffe P.J., Maxwell P.H, & Nathan J.A. (2021). The HIF complex recruits the histone methyltransferase SET1B to activate specific hypoxia-inducible genes. Nature genetics, 53(7), 1022-1035.
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5

Quantitative Analysis of Tumor Spheroid Growth

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2.5 × 103 HeLa cells were seeded in 96-well Clear Round Bottom Ultra-Low Attachment Microplates (7007, Corning®) in a final volume of 200 μl. Cells were centrifuged at 2,000 rpm for 10 min to encourage the formation of a single spheroid per well. 12 replicates of each condition were plated in each biological replicate. Every three days, 100 μl cell culture medium was replaced, and every two days spheroids were photographed in an EVOS™ M500 (Thermo Fisher Scientific™) at 4× magnification. Spheroids were cultured for 7 days. Fiji Image J Software was used to measure the volume and circularity of spheroids. Spheroid area was calculated as previously reported61 (link), and used to determine spheroid radius (R = √(area/π)), from which the volume (V = (4/3) πR3) was calculated. Each spheroid’s GFP levels were quantified using Fiji Image J Software and the intensity relative to spheroid volume was depicted.
Ortmann B.M., Burrows N., Lobb I.T., Arnaiz E., Wit N., Bailey P.S., Jordon L.H., Lombardi O., Peñalver A., McCaffrey J., Seear R., Mole D.R., Ratcliffe P.J., Maxwell P.H, & Nathan J.A. (2021). The HIF complex recruits the histone methyltransferase SET1B to activate specific hypoxia-inducible genes. Nature genetics, 53(7), 1022-1035.
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6

Evaluating Anti-Trypanosoma cruzi Activity

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The IC50 values for amastigote forms of T. cruzi were determined as follows. Peritoneal macrophages were collected from BALB/c mice and seeded (1x105) in 16-well chamber slides (NUNC, Thermo Fisher Scientific). These macrophages were infected with T. cruzi trypomastigotes (1:10 macrophage-parasite ratio) during 2 h. After washing out the extracellular parasites with medium, the infected cells were treated with the test compounds in different concentrations (60 to 0.93 μM, 1:2 dilution) at 37 °C in a 5% CO2 humidified incubator for 48 h. The cells were transferred to slides and were then fixed with MeOH, stained with Giemsa and observed under a light microscope (EVOS M500, Thermo). Investigation of compounds’ cytotoxicity to macrophages was verified at the highest concentration (i.e. 60 µM) through macrophages’ counts, and only compounds that did not caused relevant cytotoxicity at this concentration were considered in the assay. The infection index was obtained in 200 macrophages and used to calculate the IC50 through non-linear regression. DMSO was added at final concentration of 0.5% (v/v) per well and an internal control was used to confirm the lack of toxicity of the solvent for the parasites at this concentration. BZN was used as the reference drug (Romanelli et al., 2022 (link)).
Varela M.T., Romanelli M., Amaral M., Tempone A.G, & Fernandes J.P. (2023). Piperazine amides with desirable solubility, physicochemical and drug-like properties: Synthesis and evaluation of the anti-Trypanosoma cruzi activity. Saudi Pharmaceutical Journal : SPJ, 31(7), 1265-1273.
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7

Cytotoxicity Evaluation of Complexes

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HeLa cells (2 × 104 per well) were seeded into 24-well plates respectively and cultured for 24 h. In this experiment, we set up control groups, IC50 groups. In the control group, the cells were cultured then the cells were treated with complexes 1 and 2 for 24 h at IC50 concentrations. Then we determined the cell viability with Calcein-AM/PI live/dead cell co-staining kit (Solarbio, Beijing, China). The concentration of Calcein-AM to stain live cells (green channel) is 2.5 μM while that of PI for the dead cells (red channel) is 4.5 μM. The photographs were captured on an inverted fluorescence microscope (Evos M500, Invitrogen by Thermo Fisher Scientific).
Tan L.T., Shen T.X., Jiang J.Y., Zhong Y.J., Lin F.Q., Xue H., Yao Y.X., Jiang X., Shen L, & He X. (2022). Bifunctional tetrazole–carboxylate ligand based Zn(ii) complexes: synthesis and their excellent potential anticancer properties. RSC Advances, 12(52), 33808-33815.
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8

Cellular Uptake of Doxorubicin-Loaded Mesoporous Silica Nanoparticles

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MCF-7 cells were seeded in 12 well plates at a density of 1 × 104 per plate and incubated in a DMEM medium for 24 h at 37 °C. Subsequently, the culture medium was replaced with PBS solutions containing MSNs, MSN@DOX, and free DOX. After 3 h of incubation, cells were washed 3 times with PBS solution, and cellular uptake of MSN@DOX was evaluated using a fluorescence microscope (EVOS M500; Invitrogen, CA, USA).
Kim M.K., Ki D.H., Na Y.G., Lee H.S., Baek J.S., Lee J.Y., Lee H.K, & Cho C.W. (2021). Optimization of Mesoporous Silica Nanoparticles through Statistical Design of Experiment and the Application for the Anticancer Drug. Pharmaceutics, 13(2), 184.
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9

Microscopic Visualization of Porous Collagen Constructs

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After the 5-day culture period, excess media were removed from each well and constructs were dehydrated using Whatman paper. Collagen constructs were placed back into respective wells of the 24-well plate and light-microscopy images obtained using a digital imaging microscope (EVOS M500, Invitrogen—Waltham MA, USA). Images of pores within the PP Medium Pore, PP macroporous and mosquito net mesh high cell density constructs (1.5 × 106 cells/ml) were captured at 4× magnification. Qualitative observation was made comparing different regions of the mesh and the collagen matrices to examine for any structural differences.
Khader R., Whitehead-Clarke T., Mudera V, & Kureshi A. (2024). Assessment of mesh shrinkage using fibroblast-populated collagen matrices: a proof of concept for in vitro hernia mesh testing. Hernia, 28(2), 495-505.
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10

Viability Assay of 3D Cell Cultures

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Single cells were encapsulated as before for six days (intestinal) or eight days (endometrial) before the addition of Calcein AM (2 mM) and Ethidium homodimer-1 (2 mM) for 20 minutes. Images were captured using either a ZEISS confocal Laser Scanning Microscope (LSM 880) equipped with temperature (37 °C), humidity, and CO2 (5%) controls or an EVOS M500 (Invitrogen) microscope (no incubation). A 1.6 mm by 1.6 mm area and ~600 μm thick section of either Matrigel or the synthetic ECM were imaged with the confocal. With the EVOS, we captured the center of the droplet, which is ~ 1/3 of the total matrix area. The final images were processed using the ZEN blue ZEISS companion software or Fiji [52 (link)]. For time-course live/dead imaging analysis, 60 z-stacks images were taken the day of seeding (day 0) then every two days for up to ten days, using the EVOS M500 microscope and processed as above.
Hernandez-Gordillo V., Kassis T., Lampejo A., Choi G., Gamboa M.E., Gnecco J.S., Brown A., Breault D.T., Carrier R, & Griffith L.G. (2020). Fully synthetic matrices for in vitro culture of primary human intestinal enteroids and endometrial organoids. Biomaterials, 254, 120125.
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