Cells were fixed with 4% paraformaldehyde, blocked with blocking buffer (10% normal goat serum, 0.3% Triton X-100 in PBS) for 1 hour, and stained with anti-Nanog antibody (1:100 dilution) overnight at 4°C. The next day, cells were washed and stained with Alexa Fluor 488-conjugated secondary antibodies (1:1,000) (Life Technologies). The nuclei were stained with DAPI, and the slides were imaged on an EVOS FL microscope (Life Technologies).
Evos fl microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom
The EVOS FL microscope is a compact, inverted fluorescence microscope designed for live-cell imaging. It features a LED illumination system and a high-resolution camera sensor for capturing images and videos of cells and other biological samples.
Automatically generated - may contain errors
Lab products found in correlation
In situ cell death detection kit, by Roche (10 mentions)
Triton x 100, by Merck Group (6 mentions)
Bovine serum albumin (bsa), by Merck Group (6 mentions)
Axio observer z1, by Zeiss (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (5 mentions)
Paraformaldehyde (pfa), by Merck Group (4 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions)
Dmem f12, by Thermo Fisher Scientific (4 mentions)
Vectashield mounting medium containing 4 6 diamidino 2 phenylindole, by Vector Laboratories (3 mentions)
Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions)
Evos xl microscope, by Thermo Fisher Scientific (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (3 mentions)
Pimozide, by Merck Group (3 mentions)
Ultra low attachment plate, by Corning (3 mentions)
Maraviroc, by Merck Group (3 mentions)
Pannoramic 250, by 3DHISTECH (3 mentions)
227 protocols using evos fl microscope
1
Nanog Expression Analysis in Cells
2
Quantifying Immune Cell Populations in Colon Tissue
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Paraffin-embedded colon tissue sections were cut into 4-μm-thick sections. Sections were deparaffinized in xylene and rehydrated sequentially in 100, 95, and 80% ethanol solutions; antigen retrieval was carried out using 10 mM citrate buffer (Sigma-Aldrich). After washing, the sections were blocked with a blocking buffer containing 1% bovine serum albumin in PBST for 30 min. The sections were incubated overnight at 4 °C with antibodies against FOXP3 (1:50; Santa Cruz Biotechnology) or CD3 (1:50; Santa Cruz Biotechnology). After three washes, the slides were incubated with secondary antibody. The colon sections, stained with an antibody against either FOXP3 or CD3, were washed three times and incubated with fluorescein-conjugated secondary antibodies (1:200; Santa Cruz Biotechnology) or Texas red-conjugated secondary antibodies (1:200; Santa Cruz Biotechnology) for 1 h at room temperature in the dark. Colon sections, stained with antibodies against either FOXP3 or CD3, were washed three times and mounted in VECTASHIELD mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA). The samples were observed using an EVOS FL microscope (Life Technologies, Darmstadt, Germany), and the immuno-reacted cells were counted in 20 random fields per group.
3
Immunohistochemical Analysis of Colon Tissue
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Paraffin-embedded colon tissues were cut into 4-µm thick sections. Sections were deparaffinized, rehydrated, and antigen retrieval was done in 10 mM citrate buffer. Sections were washed and blocked with a buffer comprising 5% bovine serum albumin and 0.3% Triton X-100 for 1 h. The sections were incubated with CD11b-PE (1:100; Santa Cruz Biotechnology) and CD206-FITC (1:100; Biolegend, USA) antibodies at 4°C overnight. The sections were washed thrice and mounted in Vectashield mounting medium containing 4′, 6-diamidino-2-phenylindole (DAPI; Vector Laboratories, USA). The sections were examined under an EVOS FL microscope (Life Technologies, Germany), and the number of immunoreactive cells was counted in 20 random fields per group.
4
Lipid Accumulation Assay in RPE
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Accumulation of lipids was assayed with the BODIPY 493/503 lipid-specific dye (Invitrogen) on RPE cultured on 24-well plates or 8-well slide chambers. Stained RPE were imaged using an EVOS FL microscope (Life Technologies, Carlsbad, CA, USA) and the number of green fluorescent lipid aggregates was counted in diseased versus normal RPE cells.
5
Immunofluorescence Microscopy of Cells
6
Fluorescence Microscopy and FACS Analysis
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Fluorescence microscopy was performed using the EVOS® FL microscope (Life Technology, CA, United States). Cells were analyzed at the bright field and with the green fluorescent protein (GFP) (ex:470 nm/em:524 nm) and red fluorescence protein (RFP) (ex:530 nm/em:593 nm) fixed filters. For an overall analysis of RFP and GFP intensity, fluorescence was quantified using the Cytation 5 Cell Imaging Multi-Mode Reader (BioTeck, VT, United States). Fluorescence experiments were performed in triplicate and the whole experiment, starting from the pSB1C3 transformation, was performed three times on different days, accounting for technical and biological replicates.
Fluorescence-activated cell sorting (FACS) (BD FACS Canto II, BD, NJ, United States) was employed to distinguish between RFP positives and negative cells upon CRISPR-Cas9 treatment. Each measurement analyzed 30,000 events at a low flow rate. The following settings were used: an SSC voltage of 473; a FSC voltage of 398; and a PerCP-Cy5-5-A of 445 V.
Fluorescence-activated cell sorting (FACS) (BD FACS Canto II, BD, NJ, United States) was employed to distinguish between RFP positives and negative cells upon CRISPR-Cas9 treatment. Each measurement analyzed 30,000 events at a low flow rate. The following settings were used: an SSC voltage of 473; a FSC voltage of 398; and a PerCP-Cy5-5-A of 445 V.
7
Evaluating GBM Cell Motility
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To evaluate cell motility, GBM cells were seeded into 6-well culture plates. When the cells reached 90% confluence, a scratch was gently made through the cell monolayer by sterile 100 μL pipette tips, and the detached cells were washed away. The cell migration was observed and imaged under an Evos FL microscope (Life Technologies) for each condition (LCTP 7.5 μM and 10 μM) and timepoint (T0 and T24 h).
8
Quantifying GBM Cell Motility Responses
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Cell motility was assessed by plating ZAR and NULU GBM cells (5 × 104/well) in 6-well plates. At 90% confluency, a “scratch” was gently created across the cell monolayer using sterile 100 μL tips. The detached cells were washed off with PBS. The cells were treated, respectively, with AE 20 μM and TMZ 10 μM as single agents and in combination (AE 20 μM and TMZ 10 μM), and DMSO 0.3% was used for control. The ability of the cells to close the “scratch” was analyzed by acquiring images under the Evos FL microscope (Life Technologies, Thermo Fisher Scientific, San Jose, CA, USA) for each time point (T0, T24 h, T48 h, T72 h, and T96 h). The scratch area was quantified by image analysis with ImageJ 1.52 software National Institutes of Health USA.
9
Fluorescent Imaging with EVOS FL
10
Evaluating MDA-MB-231 Cell Delivery Efficiency
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