Exponentially growing NCI‐H460 cells were seeded in 12‐well plates (300 cells/well) and treated with 25 μmol/l dihydroberberine, 2 μmol/l sunitinib, or 25 μmol/l dihydroberberine plus 2 μmol/l sunitinib for 48 hrs. Subsequently, the media was changed, and the cells were cultured in drug‐free media for an additional 10–15 days, until colonies were obviously visible and countable. Then, the colonies were fixed with methanol and stained with crystal violet. Images were photographed under the chemiluminescent and fluorescent imaging system (Champchemi Professional, SG2010084, Sage creation, Beijing, China) and an inverted fluorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
Dm505
Manufactured by Nikon
Sourced in Japan
The DM505 is a digital microscope camera from Nikon. It features a high-resolution CMOS image sensor and is designed for image capture and analysis in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Ex460 500, by Nikon (5 mentions)
Ba510 560, by Nikon (5 mentions)
Eclipse ti, by Nikon (4 mentions)
Neo scmos, by Oxford Instruments (4 mentions)
Ex540 25, by Nikon (3 mentions)
Dm565, by Nikon (3 mentions)
Plan apo 60 n a 1.4 objective, by Nikon (2 mentions)
Nis elements viewer 4, by Nikon (1 mentions)
Eclipse 80i upright microscope, by Nikon (1 mentions)
Ds ri2, by Nikon (1 mentions)
Nis elements f, by Nikon (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
Eclipse te2000 u, by Nikon (1 mentions)
Dm575, by Nikon (1 mentions)
Ex 535 50, by Nikon (1 mentions)
Zyla 5, by Oxford Instruments (1 mentions)
19 protocols using dm505
1
Synergistic Anti-Cancer Effects of Dihydroberberine and Sunitinib
2
Mitochondrial and Immunofluorescence Assay
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Cells were stained with Mitotracker and then xed with 4% paraformaldehyde for 10 min and then blocked with 10% BSA for 30 min at room temperature. After this procedure, the cells were incubated with primary antibodies (1:200) at 37°C for 4 h. Then, cells were incubated with an CoraLite488-conjugated anti-rabbit secondary antibody (1:50) at room temperature for 1 h. The nucleus was stained with DAPI.
Fluorescent signals were detected using an inverted uorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
Fluorescent signals were detected using an inverted uorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
3
Investigating Cell Migration Dynamics
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After reaching 80% confluence, Bel-7402 and SMMC-7721 cell monolayers were scratched using pipette tips (10–100 μL) and treated with different stimuli (LPS: 5 μg/mL, ATP: 2 mM, 5 μg/mL LPS + 2 mM ATP) in the presence of 3% fetal bovine serum for 48 h. The migration distance of cells in each group was determined using image analysis software (NIS-Elements Viewer 4.2.0; Nikon Corporation) and calculated as the migration rate. Images of cells were captured at 0 h and 24 h after stimulation treatment using an inverted fluorescence microscope (DM505, Nikon Corp). By comparing the images taken at 0 h and 24 h and analysing the migration distances using the image analysis software, the effects of the different stimuli on cell migration can be assessed.
4
Cantharidin Cytotoxicity in Breast Cancer
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MDA-MB-231 and MDA-MB-468 cells were seeded and then treated with different concentrations of cantharidin for 48 h. MDA-MB-231 and MDA-MB-468 cells were seeded and incubated with miR-607 NC, miR-607 mimic and miR-607 inhibitor for 24 h. The medium was then replaced with drug-free medium, and the plates were cultured for an additional 10–15 days until the colonies were clearly visible and countable. Colonies were fixed with methanol and stained with crystal violet. Images were obtained using an imaging system (Champchemi Professional, SG2010084, Sage creation, Beijing, China) and an inverted fluorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
5
Visualizing Nitric Oxide in Kas Roots
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The accumulation of NO in Kas roots was visualized using a NO probe, DAF-FM DA (4-amino-5-methylamino-2,7-difluorofluorescein diacetate). First, after washing with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)-KOH (pH 7.4), the root apex was immersed in 200 μL DAF-FM DA (10 μM) for 30 min in dark. Excess fluorescence was removed by washing with HEPES-KOH (pH 7.4) three times, then visualized using an Eclipse 80i upright microscope with the following filters: EX 460-500, DM 505, and BA 510-560 (Nikon, Minato, Tokyo, Japan). The intensity of the fluorescence was calculated using Photoshop 7.0 (Adobe Systems Inc., San Jose, CA, USA).
6
Azobenzene Microscopy Imaging Protocol
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The samples were illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using an oil-coupled Plan Apo 60× N.A.1.4 objective (Nikon). UV cut-off filter blocks (TRITC: EX 540/25, DM565, BA605/55; GFP-B: EX460-500, DM505, BA510-560; Nikon) were used in the optical path of the microscope. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. Two ND filters (ND4, 25% transmittance for TRITC and ND1, 100% transmittance for GFP-B) were inserted into the illumination light path of the fluorescence microscope to reduce photobleaching of the samples. In order to isomerize the azobenzene units, the flow cell was irradiated with the light passed through a UV-1A filter block (UV-1A: EX 365-410, DM400, BA400; Nikon).
7
Immunofluorescence Staining of Fixed Cells
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Cells were fixed with 4% paraformaldehyde for 10 min and then blocked with 10% BSA for 30 min at room temperature. After this procedure, the cells were incubated with primary antibodies (1:200) at 37 °C for 4 h. Then, cells were incubated with an CoraLite488-conjugated anti-rabbit and CoraLite594-conjugated anti-mouse secondary antibody (1:50) at room temperature for 1 h. The nucleus was stained with DAPI. Fluorescent signals were detected using an inverted fluorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
8
Sanguinarine Concentration Impacts on Cell Colony Formation
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Cells were seeded in a 12-well plate (200 cells/well) and treated with different concentrations of sanguinarine for 48 h. The medium was changed to drug-free medium for additional 10–15 days. Then, the colonies were fixed with methanol and stained with 0.2% crystal violet for 15 min at room temperature. Images were taken using the white light imaging system (Champchemi Professional SG2010084, Sage Creation, Beijing, China) and the inverted fluorescence microscope (DM505, Nikon Co., Ltd., Otawara, Tochigi, Japan).
9
Visualizing Fungal Infection Stages with Fluorescence Microscopy
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A manual inverted microscope (Ti-S; Nikon, Tokyo, Japan) equipped with a Super Plan Fluor ELWD ADM 20/0.45 was used to observe the eGFP expression in mycelia, conidia, germ tube, appressorium and invasive hyphae. eGFP was exited using a HG fiber illuminator with filter cube (Excitation Filter EX450-490, Dichroic Mirror DM505, Barrier Filter BA520, Tokyo, Japan) and images were acquired using a high-resolution color camera head DS-Ri2 (Nikon, Tokyo, Japan). All parts of the system were under the control of the NIS-elements F (Nikon, Tokyo, Japan).
10
Visualizing Tubulin Microtubule Assembly
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The droplet of tubulin solution after drying and stabilizing with taxol buffer, was illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using 2×, 20× objective lens (Nikon). UV cut-off filter block (GFP-B: EX460-500, DM505, BA510-560; Nikon) was used in the optical path of the microscope. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. ND filter (ND32, 3.1% transmittance for GFP-B) was inserted into the illumination light path of the fluorescence microscope to reduce photobleaching of the samples.
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