M3/18 cells were seeded and grown overnight at 39.5 °C on glass-bottom 35-mm tissue culture dishes (MatTek) in complete growth medium. After siRNA transfection for 48 h, the cells were ready for FRAP at 39.5 °C in 5% CO2. FRAP experiments were performed on a Leica TCS SP5 confocal microscope as previously described (Nikonov et al., 2002 (link)).
35 mm glass bottom tissue culture dishes
Manufactured by MatTek
Sourced in United States
The 35-mm glass bottom tissue culture dishes are a laboratory equipment designed for cell culture applications. They provide a transparent glass surface for microscopic observation and analysis of cells in their growth environment.
Automatically generated - may contain errors
Lab products found in correlation
Tcs sp5 confocal microscope, by Leica (1 mentions)
Stage top incubator, by Okolab (1 mentions)
Ab219801, by Abcam (1 mentions)
Tcs sp8 microscope, by Leica (1 mentions)
Ixon life 888 emccd camera, by Oxford Instruments (1 mentions)
Dragonfly 505, by Leica (1 mentions)
Dmi8 microscope, by Oxford Instruments (1 mentions)
Bz x810, by Keyence (1 mentions)
Anti ki 67 8d5, by Cell Signaling Technology (1 mentions)
F ab 2 fragment alexa fluor 488 conjugate, by Cell Signaling Technology (1 mentions)
Bz x810 imaging system, by Keyence (1 mentions)
Lsm800 confocal microscope, by Zeiss (1 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
7 protocols using 35 mm glass bottom tissue culture dishes
1
Fluorescence Recovery After Photobleaching
2
Imaging HEK cells under Dox induction
3
Quantifying Cell Proliferation with EdU Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EdU cell proliferation assay was performed per manufacturer recommendation (ab219801, Abcam). Briefly, 1 × 105 cells were seeded on 35-mm glass bottom tissue culture dishes (MatTek Corporation) in growth media. At ~ 70–80% confluence, cells were incubated for 6 h with 5-ethyl-2-deoxyuridine (EdU, 10 µM), which incorporates into the newly synthesized DNA. After fixation, permeabilization, and blocking of the cells, EdU incorporation was visualized using the Click it EdU-Alexa-Flour488 imaging kit. The cells were treated with DAPI to visualize nuclei (Thermo Fisher).
4
Multimodal Live-Cell and Fixed-Cell Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 35 mm glass bottom tissue culture dishes (MatTek, USA) or in 35 mm glass bottom tissue culture dish with 4 compartments (Greiner, Austria). Image acquisition was performed by an Andor Dragonfly 505 spinning disk confocal on a Leica DMi8 microscope with an Andor iXon Life 888 EMCCD camera and Andor Zyla 4.2 + sCMOS camera using a 565 nm long pass dichroic image splitter. (Oxford, UK). The microscopy set‐up allowed for the simultaneous acquisition of two channels. For live cell imaging, the microscopes were equipped with a humidified climate control system at 37˚C supplemented with 5% CO2. Specific frame rates are noted at the relevant experiments.
For fixed cell imaging, cells were seeded on glass coverslips and transfected. The day after transfections, cells were washed with PBS and fixed in 3.7% (w/v) formaldehyde in PBS solution for 10min at RT. Next, cells were washed and mounted. Samples were imaged using a Leica TCS SP8 microscope (Leica Microsystems, Wetzlar, Germany) equipped with 405/488/561/647 nm laser lines. Images were analyzed using ImageJ Software.
For fixed cell imaging, cells were seeded on glass coverslips and transfected. The day after transfections, cells were washed with PBS and fixed in 3.7% (w/v) formaldehyde in PBS solution for 10min at RT. Next, cells were washed and mounted. Samples were imaged using a Leica TCS SP8 microscope (Leica Microsystems, Wetzlar, Germany) equipped with 405/488/561/647 nm laser lines. Images were analyzed using ImageJ Software.
5
Senescence Assay in MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WJ and BM MSCs 1 × 105 were seeded on 35 mm glass bottom tissue culture dishes (MatTek Corporation) in growth media. At ~ 70–80% confluence, cells were fixed and Beta-Gal activity was detected with a kit per manufacturer’s recommendation. Stained cells were imaged with brightfield microscopy with the Keyence BZ-X810 imaging system.
6
Cell Cycle Analysis using Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Briefly, 1 × 105 cells were seeded on 35-mm glass bottom tissue culture dishes (MatTek Corporation) in growth media. At ~ 70–80% confluence, cells were fixed, permeabilized with freeze cold methanol for 10 min, and washed in PBS. Subsequently, cells were labeled with primary antibodies: anti-phospho-Histone H3 (Ser10) (6G3) (Cell Signaling, #9706) anti-Ki-67 (8D5) (#9449, Cell Signaling). After overnight staining at 4˚C, cells were washed in PBS and stained with secondary antibody (Anti-mouse IgG (H + L), F(ab')2 Fragment [Alexa Fluor® 488 Conjugate] #4408 Cell Signaling) at room temperature. After 2 h, the cells were washed in PBS and stained with Hoechst H33342 (5 µg/mL) to visualize nuclei. Stained cells were imaged within 24 h with the Keyence BZ-X810 imaging system.
7
Visualizing PCM-1 Dynamics in RPE-1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mock‐ or EHD1‐siRNA‐treated RPE‐1 cells on 35 mm glass‐bottom tissue culture dishes (MatTek, Ashland, MA) were transiently transfected with PCM‐1‐GFP for 24 h, and then serum‐starved in DMEM/F12 with 2 mM L‐glutamine, 1× nonessential amino acids and 0.2% FBS for 15 min at 37°C. The cells were imaged in prewarmed Opti‐MEM medium (Gibco) containing 2 mM L‐glutamine with a Zeiss LSM 800 confocal microscope (Carl Zeiss, Jena, Germany). Images were obtained every 5 s for 10 min with the pinhole set to obtain z‐sections of 1 μm. Images and videos were cropped, adjusted for brightness (whole‐image adjustment), and time‐stamped with minimal manipulation for presentation. For quantification, we performed three independent experiments. pEGFP‐N‐PCM1 (Dammermann & Merdes, 2002 (link)) was a kind gift from Dr. Andreas Merdes (Université Toulouse III, Toulouse, France).
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!