H9c2 cell proliferation was evaluated using the 5-ethynyl-2′-deoxyuridine (EdU) method. Cultured H9c2 cells (1×105/ml) were stained at 37°C with 1 µM EdU solution (Thermo Fisher Scientific, Inc.) for 30 min, incubated with 1X Hoechst 33342 solution (Beyotime Institute of Biotechnology) for 10 min at room temperature, and observed using a fluorescence microscope (magnification, ×200). The apoptosis of H9c2 cells was analyzed by staining cells with 5 µg/ml Hoechst 33258 for 20 min at room temperature, and observed with a confocal microscope (magnification, ×120). H9c2 cell apoptosis analysis was performed using flow cytometry using the Annexin V-FITC Apoptosis Staining/Detection kit (cat. no. ab14085; Abcam), and cells were incubated with 5 µl Annexin V-FITC and 5 µl propidium iodide at room temperature for 5 min in the dark, and analyzed using WinMDI software (Version 2.8, J.; Trotter, Scripps Research Institute). H9c2 cell migration was evaluated using the Transwell system-based assay. H9c2 cells (5×105/ml) resuspended in serum-free DMEM were seeded and cultured in the upper chamber for 24 h. FBS (10%) in DMEM was used as a chemoattractant. The migrated cells were finally observed using a light microscope (TE200, Nikon Corporation; magnification, ×200) by staining with 0.1% crystal violet at RT for 15 min.
Te200
Manufactured by Nikon
Sourced in Japan, United States, United Kingdom
The Nikon TE200 is a microscope designed for laboratory use. It features a rugged and stable construction to provide reliable performance in various research and analysis applications. The TE200 offers essential microscope functions to support your scientific work.
Automatically generated - may contain errors
Lab products found in correlation
Softworx imaging software, by Cytiva (3 mentions)
Oil red o solution, by Merck Group (3 mentions)
Mitosox red, by Thermo Fisher Scientific (3 mentions)
Deltavision deconvolution microscope, by Cytiva (2 mentions)
Coolsnap hq, by Teledyne (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Fura 2 acetoxymethyl ester, by Thermo Fisher Scientific (2 mentions)
C1sir, by Nikon (2 mentions)
Hoechst 33342 solution, by Beyotime (1 mentions)
Edu solution, by Thermo Fisher Scientific (1 mentions)
B8434, by Merck Group (1 mentions)
Softworx software, by Cytiva (1 mentions)
Deltavision deconvolution system, by Cytiva (1 mentions)
Metamorph, by Universal Imaging (1 mentions)
Cm h2dcfda, by Thermo Fisher Scientific (1 mentions)
Fluorescence microplate reader, by Molecular Devices (1 mentions)
Metamorph, by Molecular Devices (1 mentions)
G box imaging system, by Syngene (1 mentions)
Deltavision microscopy system, by Cytiva (1 mentions)
Softworx program, by Cytiva (1 mentions)
65 protocols using te200
1
Evaluating H9c2 cell proliferation, apoptosis, and migration
2
Foam Cell Formation Assay in RAW264.7
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Foam cell assay was carried out in RAW264.7 cell as described previously with slight modification [24 (link)]. Oil Red O (ORO) staining was performed in oxidized-low density lipoprotein (ox-LDL) and/or baicalin with/without 10 μg/mL of MØ-MP treated RAW264.7 cell. Positive stained (red) foam cells were observed via phase-contrast microscope (TE200; Nikon, Tokyo, Japan). Photographs were taken at 40 × magnification. Colorimetric quantification of lipid content in oxidized LDL and/or MØ-MP treated RAW 264.7 cells after ORO staining was performed.
3
Intercellular Communication in HeLa-Cx43 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The intercellular communication via gap junctions between HeLa-Cx43 cells (ATCC) was evaluated in subconfluent cultures (85%) by iontophoretic injection into one cell of 5% wt/vol Lucifer yellow (150 mM) in HCO−3 free F-12 medium buffered with 10 mM HEPES, pH 7.2 through glass microelectrodes. Briefly, coverslips containing cells were placed in a perfusion chamber and visualized in an inverted microscope (TE 200; Nikon, Melville, NY) equipped with xenon arc lamp and filters for Lucifer yellow (excitation wavelength 450–490 nm; emission wavelength above 520 nm). At 1 min after dye injection, surrounding cells were examined to determine whether dye transfer occurred. The incidence of dye coupling was calculated as the percentage of cases in which the dye transferred to at least one adjacent cell and the number of cells to which dye spread was determined and expressed as the index of dye coupling. In all experiments, the incidence of dye coupling was evaluated by injecting a minimum of ten cells.
4
Silicone Oil-Alginate Emulsion Droplet Sizing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The droplet size distributions were measured for the silicone oil-alginate solution emulsions prepared from: 10 cSt silicone oil stirred by a magnetic stir bar, 10 cSt silicone oil stirred by an overhead disk turbine and a 500 cSt silicone oil stirred by a magnetic stir bar. Samples from the emulsions were diluted by a 0.5 wt% alginate solution with TX-100 in order to properly visualize individual drops. Images were recorded on an inverted microscope (Nikon TE-200, Melville, NY, USA) using an attached CMOS camera (MotionPro X3, Tallahassee, FL, USA). The images were then analyzed using the ImageJ particle analysis tool.
5
BrdU Incorporation Assay for Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated onto coverslips, subjected to the indicated treatments and then BrdU (10 mM) was added and cells were incubated for additional 3 h. After fixed in 4% paraformaldehyde and permeabilized with Triton X-100, cells were washed three times with PBS for 5 min and DNA was denaturalized by incubating 10 seconds in 50 mM NaOH. After treatment with a blocking solution, the cells were incubated with anti-BrdU antibody (B8434 Sigma Chemical Co.), washed and incubated with secondary antibody, washed once again and incubated with 4′,6-diamidino-2-phenylindole (DAPI, Molecular Probes) for nuclei staining. The cells were then washed and mounted for observation under a fluorescence microscope (Nikon TE200). Independent areas were captured and BrdU-positive nuclei were counted and expressed as percentage (a total of 500 nuclei on average were counted).
6
Wound Healing Assay in U2OS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Confluent U2OS monolayers, grown in each chamber of a LabTek 8-chamber glass slide in DMEM (Mediatech) supplemented with 10% fetal bovine serum, were wounded with a pipette tip, and filming was initiated 1 h after wounding. Low magnification phase-contrast videos were taken at 37°C using an inverted microscope (TE-200; Nikon) equipped with a 20× Plan Fluor phase lens (NA 0.5), a 12-bit chilled charge-coupled device (CCD) camera (C4742-95 12G04; Hamamatsu Photonics), and a robotic XYZ microscope stage (MS-2000; Applied Scientific Instruments, Inc.) equipped with linear positional feedback controllers (Haidenhain) on all three axes. Images (200-ms exposure) were collected at 7-min intervals for 24 h. Up to 30 fields were simultaneously counted in each experiment. Image stacks were generated in MetaMorph (Universal Imaging Corp.) and then converted to 14-s QuickTime (Apple) videos at 17 frames/s. For Fig. 7 and Fig. S1, cells were grown in Liebovitz’s L-15 medium (Mediatech) supplemented with 10% fetal bovine serum, and live-cell images were captured in differential interference contrast (DIC) at 37°C with an inverted microscope (IX81; Olympus) using an LC Plan FIuor 20×/NA 0.40 objective, within a DeltaVision Deconvolution system (Applied Precision) equipped with a CCD camera (CoolSNAP; Photometrics). The images were acquired with softWoRx software (Applied Precision).
7
Measuring Intracellular ROS and Superoxide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular levels of reactive oxygen species production were measured as described previously (Wei et al., 2000). Detection of mitochondrial superoxide was measured using the fluorogenic probe, MitoSOX™ Red (Invitrogen, Carlsbad, CA, USA) (Won et al., 2015). bEnd.3 cells were subjected to OGD for 4 hours, then switched to a reperfusion condition and incubated for 60 minutes. Cells were loaded with 5 μM CM-H2DCFDA (Invitrogen) at 37°C for 10 minutes (reactive oxygen species levels), or 5 μM MitoSOXTM Red (Invitrogen) at 37°C for 30 minutes (superoxide production). Thereafter, cells were washed three times with PBS to remove residual probe. Cellular fluorescence intensity was measured at ex/em wavelengths of 488/525 nm and 510/580 nm, respectively, using a fluorescence microplate reader (Molecular Devices). Expression of fluorescence intensity was expressed as percentage of the control group. Images were captured using an inverted fluorescence microscope (TE200; Nikon, Tokyo, Japan).
8
Fluorescence Microscopy for Nuclear Fusion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All fluorescence microscopy was performed on a DeltaVision deconvolution microscope (Applied Precision, Issaquah, WA), based on a Nikon TE200 (Melville, NY) with an inverted 100X NA 1.4 objective, a 50-W mercury lamp, and a Photometrics Cool Snap HQ CCD camera (Photometrics, Tucson, AZ). Image pixel size is 49.2 × 49.2 nm. All images were deconvolved using the Applied Precision SoftWoRx imaging software.
For nuclear fusion assay microscopy (fixed, DAPI-stained cells), we acquired large z-stacks that captured the entire cell (typically 19 slices separated by 0.2 μm). The remaining imaging experiments used live cells in growth medium with smaller z-stacks (typically ~9 slices separated by 0.2 μm) to avoid photobleaching.
For nuclear fusion assay microscopy (fixed, DAPI-stained cells), we acquired large z-stacks that captured the entire cell (typically 19 slices separated by 0.2 μm). The remaining imaging experiments used live cells in growth medium with smaller z-stacks (typically ~9 slices separated by 0.2 μm) to avoid photobleaching.
9
Histological Analysis of Mouse Organs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After dewaxing and dehydration, sections of lung, liver and kidney tissues of mice were soaked in Harris hematoxylin for 3–8 min, hydrolyzed with 1% hydrochloric acid alcohol, and treated with 0.6% ammonia to return to blue. Sections were stained with eosin for 1–3 min, and then immersed in 95% alcohol I and II, anhydrous ethanol I and II each for 5 min, and in xylene I and II each for 5 min for dehydration and cleaning purposes. Sections were sealed with neutral gum and examined under a microscope (Nikon, Tokyo, Japan, TE200). The images were collected to analyze lung, liver, and kidney pathology.
10
Microscopic Analysis of Meiotic Spindle Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For microscopy, 100 μl of a sporulated culture expressing GFP-TUB1 integrated at TUB1 (Straight, Marshall et al. 1997 (link)) and SPC42-mCherry was spun down and resuspended in 10 μl of 1% potassium acetate media. Cells were imaged on a DeltaVision deconvolution microscope (Applied Precision, Issaquah, WA) based on a Nikon TE200 (Melville, NY) using a 100x/numerical aperture 1.4 objective, a 50W mercury lamp, and a Photometrics Cool Snap HQ charge-coupled device camera (Photometrics, Tucson, AZ). All images were deconvolved using Applied Precision SoftWoRx imaging software. At least one hundred cells were counted in all cases, and the percentage of cells in each meiotic stage including mature spores were determined using the morphology of the spindle and number of spindle pole bodies.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!