35 mm glass bottom petri dishes

Manufactured by Cellvis

Sourced in United States

The 35-mm glass bottom petri dishes are a type of cell culture dish with a thin glass bottom. They are designed to provide a transparent surface for microscopic observation of cells or other biological samples.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Mccoy s 5a, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Ht 29 cell, by American Type Culture Collection (1 mentions) Penicillin streptomycin, by Sartorius (1 mentions) Hyclone, by Thermo Fisher Scientific (1 mentions) Ixon 888 u3 ultra em ccd, by Oxford Instruments (1 mentions) 2 aminoethoxydiphenyl borate 2 apb, by Bio-Techne (1 mentions) Imageem, by Hamamatsu Photonics (1 mentions) 1 6 hexanediol, by Merck Group (1 mentions)

7 protocols using 35 mm glass bottom petri dishes

1,6-Hexanediol Neuronal and HEK Cell Experiments

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For 1,6-Hexanediol (Sigma) experiments in neurons, transfected neurons were treated immediately prior to imaging. Neurons plated on 25-mm coverslips were mounted onto a metal ring sample holder containing 3/4 of the final volume of cell culture media. Upon initiation of the imaging experiment, the remaining 1/4 cell culture media was added from a stock solution that contains 28% 1,6 Hexanediol to make a final concentration of 7% 1,6 Hexanediol.
For HEK293T cells, cells were plated onto 35-mm glass bottom petri dishes (Cellvis). The stock solution was 16%, for a final concentration of 4%. Dishes contained 3/4 of the final volume of cell culture media. Upon initiation of the imaging experiment, the remaining 1/4 cell culture media was added from a stock solution that contains 16% 1,6 Hexanediol to make a final concentration of 4% 1,6 Hexanediol.
For HEK293 cell experiments, pre-warmed 1,6-Hexanediol was diluted to final concentration of 3% in DMEM (culture media) and loaded onto cells plated on 35-mm glass bottom petri dishes (Cellvis, US).

Ogunmowo T., Hoffmann C., Pepper R., Wang H., Gowrisankaran S., Ho A., Raychaudhuri S., Cooper B.H., Milosevic I., Milovanovic D, & Watanabe S. (2023). Intersectin and Endophilin condensates prime synaptic vesicles for release site replenishment. bioRxiv.

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HT-29 Cells Arid1a Knockdown

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HT-29 cells (American Type Culture Collection, Manassas, VA) were grown in Gibco formulated McCoy’s 5A medium (Life Technologies, Carlsbad, CA) supplemented with 10% (v/v) FBS (Sigma-Aldrich, St. Louis, MO) and grown at 37°C and 5% CO₂. All cells in this study were maintained between passage 5 and 25. Transient HT-29 Arid1a short hairpin RNA knockdown line (A-Kd) was produced using a Lipofectamine vector. Quantitative reverse transcription PCR was used to assess for knockdown: Imaging and microarrays were performed on clones that demonstrated at least an 80% reduction in Arid1a expression compared to the control vector.
Before imaging, cells were cultured in 35-mm glass bottom petri dishes (Cellvis, Mountain View, CA) until at least 50% confluent. Cells were given at least 24 hours to readhere before 5 hours of serum deprivation. For serum deprivation, cells were grown in fresh McCoy’s 5A (Life Technologies) without serum supplementation and maintained at 37°C and 5% CO₂.

Virk R.K., Wu W., Almassalha L.M., Bauer G.M., Li Y., VanDerway D., Frederick J., Zhang D., Eshein A., Roy H.K., Szleifer I, & Backman V. (2020). Disordered chromatin packing regulates phenotypic plasticity. Science Advances, 6(2), eaax6232.

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Cell Culture Protocol for Imaging

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A549, COS-7, and HeLa cell lines were purchased from the Shanghai Institute of Biological Sciences (Shanghai, China). All cells were cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone, Thermo Scientific) supplemented with 10% fetal bovine serum (Bioind), antibiotics (Bioind), and penicillin/streptomycin (Bioind) at 37°C and 5% CO2. The cultured cells were passaged every 2 days. For confocal imaging, cells were plated in 35 mm glass-bottom petri dishes (Cellvis).

Xu H., Zhang J., Zhou Y., Zhao G., Cai M., Gao J., Shao L., Shi Y., Li H., Ji H., Zhao Y, & Wang H. (2022). Mechanistic Insights into Membrane Protein Clustering Revealed by Visualizing EGFR Secretion. Research, 2022, 9835035.

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Live-cell confocal imaging and 1,6-Hexanediol assay

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Live-cell confocal imaging and the 1,6-Hexanediol assay in HEK293 cells were performed as described previously^{71 (link)}. In short, HEK293 cells were plated onto 35-mm glass bottom petri dishes (Cellvis, US) and transfected with plasmids as indicated in the text using Lipofectamine 2000 (Thermo Fisher). Pre-warmed 1,6-Hexanediol was diluted to final concentration of 3% in DMEM (culture media) and loaded to cells. Imaging was performed on the Eclipse Ti Nikon Spinning Disk Confocal CSU-X, equipped with OkoLab Live-cells incubator (for control of temperature at 37C°, 5% CO2), 2 EM-CCD cameras (AndorR iXon 888-U3 ultra EM-CCD), Andor Revolution SD System (CSU-X), objectives PL APO 60x/1.4 NA oil immersion lens. Excitation wave lengths were: 405-nm for BFP, 488-nm for GFP; 561-nm for mCherry, 640-nm for emiRFP. All images were analyzed with ImageJ (NIH).

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Measuring Intracellular Calcium Dynamics in MEC

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MEC were seeded on 35 mm glass bottom petri dishes (Cellvis, Sunnyvale, CA, USA) at a density of 5000 cells per dish. They were loaded with the intracellular Ca²⁺ indicator fura-2/AM (0.5 µM) for 1 hour in the dark. Then cells were stimulated with OXT (10⁻⁸, 10⁻⁷, and 10⁻⁶ M). In selected experiments, cells were preincubated for 20 minutes with the IP₃ receptor antagonist, 2-aminoethoxydiphenyl borate (2-APB; 10⁻⁴ M; Tocris Bioscience, Bristol, UK) before addition of OXT. UTP (10⁻⁵ M) was used as a positive control in each experiment. Changes in intracellular [Ca²⁺] were recorded using a ratio imaging system (InCyt Im2; Intracellular Imaging, Cincinnati, OH, USA).⁷ (link) As fura-2 binds to free calcium in the cell, its peak absorption wavelength changes from 380 nm (unbound) to 340 nm (bound), whereas the emission wavelength remains at 510 nm. The 510 nm emissions are captured by the camera as black and white images and the ratio of emission intensity excited by 340 nm and 380 nm light is recorded. These ratios are compared to a standard curve derived from standard calcium solutions to determine ion concentration. The use of ratios ensures that measurements are not changed by variable dye concentration or cell thickness.

Gárriz A., Aubry S., Wattiaux Q., Bair J., Mariano M., Hatzipetrou G., Bowman M., Morokuma J., Ortiz G., Hamrah P., Dartt D.A, & Zoukhri D. (2021). Role of the Phospholipase C Pathway and Calcium Mobilization in Oxytocin-Induced Contraction of Lacrimal Gland Myoepithelial Cells. Investigative Ophthalmology & Visual Science, 62(14), 25.

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Quantifying zMADM-labeled Cells in Zebrafish Larvae

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The 24-hpf zMADM embryos were manually dechorionated and treated with 0.004% PTU in egg water to reduce pigmentation and raised to 4 dpf. Then larvae were fixed with 4% paraformaldehyde (PFA) at 4 °C overnight. On the next day, fixed larvae were washed with 0.3% PBT three times and stained with 1 μg/mL DAPI in 0.3% PBT (PBS containing 0.3% Triton-X 100) for 2 h at room temperature in the dark. After washing three times with 0.3% PBT, the larvae were immersed in 0.8% low-melting-point agarose and mounted in 35-mm glass-bottom Petri dishes (Cellvis) for imaging. For each larva, three optic slices were taken to quantify the total number of DAPI- and zMADM-labeled cells. The ImageJ plugin, Image-based Tool for Counting Nuclei (ITCN), was used to count the DAPI number with 15 pixels in width and 6 pixels in minimum distance. zMADM-labeled cells were manually counted. Labeling efficiency was determined by dividing zMADM-labeled cells by total DAPI number.

Xu B., Kucenas S, & Zong H. (2022). zMADM (zebrafish mosaic analysis with double markers) for single-cell gene knockout and dual-lineage tracing. Proceedings of the National Academy of Sciences of the United States of America, 119(9), e2122529119.

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Probing Chromatin Structure in hMSCs

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Bone Marrow-Derived Mesenchymal Stromal Cells (hMSCs) were cultured in 35 mm glass-bottom Petri dishes (Cellvis, Mountain View, CA) with a micropatterned or flat mPOC surface in growth medium or osteogenic differentiation medium at 37°C and 5% CO2. The PWS microscopy images were acquired on a commercial inverted microscope (Leica, Buffalo Grove, IL, DMIRB) with a Hamamatsu Image EM charge-coupled device camera (C9100-13) coupled to a liquid crystal tunable filter (CRi, Woburn, MA) to collect spectrally resolved images between 500 to 700 nm with 1 nm step size. Further, broadband illumination is provided by an Xcite-120 LED lamp (Excelitas, Waltham, MA). PWS microscopy was used to capture spatial variations of the refractive index distribution or chromatin packing density heterogeneity (

\sum

) within the nucleus. Further, the statistical parameter of chromatin structure, packing scaling (

D

) was calculated from

\sum

.^{31 (link)} At least 10 independent fields of view were utilized for each experiment and four biological replicates were used for the analysis.

D

was calculated for 111 hMSCs from the flat surface, and 110 hMSCs on micropillar surfaces.

Wang X., Agrawal V., Dunton C.L., Liu Y., Virk R.K., Patel P.A., Carter L., Pujadas E.M., Li Y., Jain S., Wang H., Ni N., Tsai H.M., Rivera-Bolanos N., Frederick J., Roth E., Bleher R., Duan C., Ntziachristos P., He T.C., Reid R.R., Jiang B., Subramanian H., Backman V, & Ameer G.A. (2023). Chromatin reprogramming and bone regeneration in vitro and in vivo via the microtopography-induced constriction of cell nuclei. Nature biomedical engineering, 7(11), 1514-1529.

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