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35 mm glass bottom culture dishes

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35-mm glass-bottom culture dishes are a type of cell culture vessel designed for microscopic observation. They feature a thin glass bottom that allows for the use of high-magnification objectives and high-resolution imaging of cells or other specimens grown in the dish.

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Lab products found in correlation

Lsm 710, by Zeiss (6 mentions) Fugene hd, by Promega (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions) Lsm 880, by Zeiss (3 mentions) Dmem f12 medium, by Fujifilm (3 mentions) Sh sy5y cell, by American Type Culture Collection (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Tcs sp8, by Leica camera (2 mentions) Tcs sp8 x confocal microscope, by Leica camera (2 mentions) Calcein green, by Thermo Fisher Scientific (2 mentions) Hcx pl apo cs 10x 0.40 dry objective, by Leica camera (2 mentions) Lasx 3, by Leica camera (2 mentions) Collagen 1, by Thermo Fisher Scientific (2 mentions) Matrigel, by BD (2 mentions) Electron multiplying charge coupled device digital camera, by Hamamatsu Photonics (2 mentions) Metamorph, by Molecular Devices (2 mentions) Vthawk two dimensional array laser scanning confocal microscopy system, by Visitech (2 mentions) Live dead biofilm viability kit, by Thermo Fisher Scientific (2 mentions)

72 protocols using 35 mm glass bottom culture dishes

1

Lipid Visualization Assay in Huh-9-13 Cells

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Nile Red was obtained from Sigma. Monodansylcadaverine
and LipidTOX Red Neutral lipid stain was obtained from life technologies.
Huh-9-13 cells were seeded in glass bottom 35 mm culture dishes (Mattek)
at a density of 25.000 cells/dish in complete DMEM without G418 at
37 °C and 5% CO2. The next day medium was replaced
with complete DMEM containing either the proper dilution of the compound
to be tested or DMSO. The cells were further incubated for 3 days
at 37 °C and 5% CO2, after which the medium was replaced
with complete medium without neutral red and either Nile Red (2 μg/mL),
monodansylcadaverine (50 μM), or LipidTOX Red Neutral lipid
stain (diluted 1/1000). Next, the cells were incubated for an additional
30 min at 37 °C and 5% CO2 after which the cells were
imaged using a laser-scanning SP5 confocal microscope (Leica Microsystems)
equipped with a DMI 6000 microscope and an Acousto optical beam splitter.
For Nile Red the yellow-gold fluorescence was monitored at an excitation
wavelength (λex) of 552 nm and an emission wavelength
(λem) of 636 nm. The red fluorescence was monitored
at λex of 485 nm and λem of 525
nm. Monodansylcadaverine was imaged at λex of 340
nm and λem of 530 nm, and LipidTOX Red Neutral lipid
stain was assessed at λex of 577 nm and λem of 609 nm.
Manfroni G., Manvar D., Barreca M.L., Kaushik-Basu N., Leyssen P., Paeshuyse J., Cannalire R., Iraci N., Basu A., Chudaev M., Zamperini C., Dreassi E., Sabatini S., Tabarrini O., Neyts J, & Cecchetti V. (2014). New Pyrazolobenzothiazine Derivatives as Hepatitis C Virus NS5B Polymerase Palm Site I Inhibitors. Journal of Medicinal Chemistry, 57(8), 3247-3262.
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2

Fluorescence Recovery Quantification by Confocal Microscopy

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Live cells were grown on glass-bottom 35 mm culture dishes (MatTek Corporation) and visualized by confocal microscopy (Zeiss LSM880 with Airyscan, Carl Zeiss Microscopy). Bleached areas were subjected to a 100% argon laser pulse at 488 nm and imaged for the indicated times post-bleaching. For A-body-detained proteins, a partial region of the A-body was photobleached, while diffuse nucleoplasmic or cytoplasmic proteins had an equal size portion of those regions photobleached instead. Intensity measurements were made using ImageJ 1.52a (National Institutes of Health) and normalized21 (link),22 ,31 (link). All fluorescence recovery after photo- bleaching experiment data represent an average of 10 cells in three independent replicates. The data was normalized69 (link) and presented as the mean of three biological replicates +/- standard error of the mean.
Marijan D., Momchilova E.A., Burns D., Chandhok S., Zapf R., Wille H., Potoyan D.A, & Audas T.E. (2024). Protein thermal sensing regulates physiological amyloid aggregation. Nature Communications, 15, 1222.
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3

Cellular Uptake of RGD-Cy5.5-Labeled Microbubbles

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Human MDA-MB-231 breast cancer cells were provided by the Stem Cell Bank, Chinese Academy of Sciences and were maintained in DMEM with 5% FBS and 1% penicillin-streptomycin at 37°C with 5% CO2. For the cellular uptake study, MDA-MB-231 cells were cultured in 35 mm glass-bottom culture dishes (MatTek, USA) with 2x104 cells mL−1 for 24 h. The medium was then replaced by a new medium containing free RGD-Cy5.5-MBs or nontargeted MBs. After 2 h of incubation, the cells were washed three times with PBS and fixed with 4% paraformaldehyde for 20 minutes. DAPI was used to stain the nuclei for 5 minutes. Fluorescence images were acquired by confocal laser scanning microscopy (TCS SP8, Leica, Germany).
Nie Z., Luo N., Liu J., Zhang Y., Zeng X, & Su D. (2020). Dual-Mode Contrast Agents with RGD-Modified Polymer for Tumour-Targeted US/NIRF Imaging. OncoTargets and therapy, 13, 8919-8929.
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4

3D Invasion Assay for Cancer Cells

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50,000 cells were embedded in a 50:50 mix of growth factor-reduced Matrigel (BD Biosciences) and collagen I (Life Technologies) and seeded onto 35 mm glass bottom culture dishes (MatTek). Cells were allowed to invade for 72h, and then they were stained with calcein green (Life Technologies) for 1h, washed with PBS and immediately imaged. Imaging was performed on a Leica TCS SP8 X confocal microscope equipped with a White Light Laser and a HCX PL APO CS 10x/0.40 DRY objective. Images were acquired as three-dimensional scans with 10 μm Z-steps and processed with LAS X 3.3 software (Leica) to obtain maximum projection images. Quantification of invasive area and invasive distance was performed on the maximum projection images using FIJI 2.3.1 distribution of ImageJ2.3.0.
Altea-Manzano P., Doglioni G., Liu Y., Cuadros A.M., Nolan E., Fernández-García J., Wu Q., Planque M., Laue K.J., Cidre-Aranaz F., Liu X.Z., Marin-Bejar O., Van Elsen J., Vermeire I., Broekaert D., Demeyer S., Spotbeen X., Idkowiak J., Montagne A., Demicco M., Alkan H.F., Rabas N., Riera-Domingo C., Richard F., Geukens T., De Schepper M., Leduc S., Hatse S., Lambrechts Y., Kay E.J., Lilla S., Alekseenko A., Geldhof V., Boeckx B., de la Calle Arregui C., Floris G., Swinnen J.V., Marine J.C., Lambrechts D., Pelechano V., Mazzone M., Zanivan S., Cools J., Wildiers H., Baud V., Grünewald T.G., Ben-David U., Desmedt C., Malanchi I, & Fendt S.M. (2023). A palmitate-rich metastatic niche enables metastasis growth via p65 acetylation resulting in pro-metastatic NF-κB signaling. Nature cancer, 4(3), 344-364.
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5

3D Invasion Assay for Cancer Cells

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50,000 cells were embedded in a 50:50 mix of growth factor-reduced Matrigel (BD Biosciences) and collagen I (Life Technologies) and seeded onto 35 mm glass bottom culture dishes (MatTek). Cells were allowed to invade for 72h, and then they were stained with calcein green (Life Technologies) for 1h, washed with PBS and immediately imaged. Imaging was performed on a Leica TCS SP8 X confocal microscope equipped with a White Light Laser and a HCX PL APO CS 10x/0.40 DRY objective. Images were acquired as three-dimensional scans with 10 μm Z-steps and processed with LAS X 3.3 software (Leica) to obtain maximum projection images. Quantification of invasive area and invasive distance was performed on the maximum projection images using FIJI 2.3.1 distribution of ImageJ2.3.0.
Altea-Manzano P., Doglioni G., Liu Y., Cuadros A.M., Nolan E., Fernández-García J., Wu Q., Planque M., Laue K.J., Cidre-Aranaz F., Liu X.Z., Marin-Bejar O., Van Elsen J., Vermeire I., Broekaert D., Demeyer S., Spotbeen X., Idkowiak J., Montagne A., Demicco M., Alkan H.F., Rabas N., Riera-Domingo C., Richard F., Geukens T., De Schepper M., Leduc S., Hatse S., Lambrechts Y., Kay E.J., Lilla S., Alekseenko A., Geldhof V., Boeckx B., de la Calle Arregui C., Floris G., Swinnen J.V., Marine J.C., Lambrechts D., Pelechano V., Mazzone M., Zanivan S., Cools J., Wildiers H., Baud V., Grünewald T.G., Ben-David U., Desmedt C., Malanchi I, & Fendt S.M. (2023). A palmitate-rich metastatic niche enables metastasis growth via p65 acetylation resulting in pro-metastatic NF-κB signaling. Nature cancer, 4(3), 344-364.
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6

Cellular Uptake and Localization of DOX Nanocarriers

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The cellular uptake and intracellular localization of DOX@NCMs and DOX@CCMs were investigated by a fluorescent microscope using cancer (HeLa) and normal (HaCaT) cells. Both cell lines were grown in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin, 1% glutamine, and 1% sodium pyruvate at 37 °C and 5% CO2. For cellular uptake experiments, fully grown HeLa and HaCaT were trypsinized and seeded into 35 mm glass-bottom culture dishes (MatTek Corporation) at a density of 2 × 105 cells/well in complete DMEM and maintained in a humidified incubator at 37 °C under 5% CO2 for 24 h. Then, 3 µg/mL of free DOX, DOX@NCMs, and DOX@CCMs (at equivalent DOX concentrations) were added to different treatment groups. After 3 and 9 h, cells were rinsed with PBS, stained with DAPI (300 nm) for 15 min, fixed with 4% paraformaldehyde solution, and the cellular uptake and localization of free DOX, DOX@NCMs, and DOX@CCMs were observed using a fluorescent microscope [3 (link)].
Birhan Y.S., Darge H.F., Hanurry E.Y., Andrgie A.T., Mekonnen T.W., Chou H.Y., Lai J.Y, & Tsai H.C. (2020). Fabrication of Core Crosslinked Polymeric Micelles as Nanocarriers for Doxorubicin Delivery: Self-Assembly, In Situ Diselenide Metathesis and Redox-Responsive Drug Release. Pharmaceutics, 12(6), 580.
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7

Fluorescence Imaging of Bacterial Cells

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Cell cultures grown in minimal medium with succinate as carbon source were incubated for 3 h to mid-exponential phase (A600 = 0,3). 0.5 ml each culture were incubated on 35-mm glass bottom culture dishes (MatTek) for 15 min, washed twice gently with phosphate sodium buffer 1X and fixed with ρ-formaldehyde (PFA) 4% during 10 min. Preparations were visualised by confocal laser scanning ZEISS LSM 880 (Carl Zeiss AG, Oberkochen, Germany) equipped with an Airyscan detection unit. To maximize the resolution enhancement a 63 × /1.46 NA oil immersion lens were used. An argon laser, at 488 nm, was used as the excitation source for the fluorescent probe. Maximum intensity projections of 5 z–stacks were analysed using ImageJ (Schneider, Rasband, & Eliceiri, 2012).Image preprocessing comprised linear adjustments as brightness and constrast followed by applying a Gaussian blur filter radius 1.5 and subsequently by a sharpening algorithm.
Monteagudo-Cascales E., García-Mauriño S.M., Santero E, & Canosa I. (2019). Unraveling the role of the CbrA histidine kinase in the signal transduction of the CbrAB two-component system in Pseudomonas putida. Scientific Reports, 9, 9110.
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8

Visualizing Cellular Internalization of Nanoplexes

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Imaging was performed using a Nikon 1AR Ultra‐Fast Spectral Scanning Confocal Microscope equipped with an environmental chamber providing temperature control. Caov3 cells were passaged onto 35 mm glass‐bottom culture dishes (MatTek, no. 0). Adherent cells were washed with DPBS and concurrently incubated with nanoplexes (50 nM, AllStars Negative Control siRNA, Alexa Fluor 488; Qiagen), LysoTracker Deep Red (50 nM, Thermo Fischer), and Hoechst 34580 (10 μg/ml, Thermo Fischer) in Opti‐MEM media at 37°C for 1 hr. Cell monolayers were then washed in DPBS and imaged in HEPES buffer (10 mM, pH 7.4).
Dreaden E.C., Kong Y.W., Quadir M.A., Correa S., Suárez‐López L., Barberio A.E., Hwang M.K., Shi A.C., Oberlton B., Gallagher P.N., Shopsowitz K.E., Elias K.M., Yaffe M.B, & Hammond P.T. (2018). RNA‐Peptide nanoplexes drug DNA damage pathways in high‐grade serous ovarian tumors. Bioengineering & Translational Medicine, 3(1), 26-36.
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9

Live-cell Imaging of Transfected Cells

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Cells were seeded on 35-mm glass-bottom culture dishes (MatTek, MA, USA) at a concentration of 2 × 105 per dish one day before imaging. The cells were incubated for 4 h with Effectene transfection reagent (Qiagen) containing 0.5 μg/dish DNA plasmid. Hoechst 33342 was added to a final concentration of 100 ng/ml to stain DNA 15 min before the removal of the transfection reagent. After the removal of the transfection reagents, the cells were washed with the prewarmed culture medium twice, and subjected to time-lapse live-cell imaging as described below. For immunofluorescence staining, CLEM and iCLEM analyses, the cells were fixed with the fixative as described in the corresponding Method sections.
Haraguchi T., Koujin T., Shindo T., Bilir Ş., Osakada H., Nishimura K., Hirano Y., Asakawa H., Mori C., Kobayashi S., Okada Y., Chikashige Y., Fukagawa T., Shibata S, & Hiraoka Y. (2022). Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase. Communications Biology, 5, 78.
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10

HepG2 Cell Culture Protocol

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EXAMPLE 15

Cell Culture: HepG2 cells were grown in Eagle's Minimum Essential Medium (EMEM) (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal calf serum (Atlanta Biologicals, Lawrenceville, Ga., USA). Penicillin/streptomycin (1%) was also present in the culture media (Invitrogen). The cells were trypsinized and collected by centrifugation, and then the cell pellet was resuspended in suitable media. An aliquot (1 mL) of the cell suspension was transferred to 35-mm glass-bottom culture dishes (MatTek Corp., Ashlan, Mass., USA) and the cells was allowed to incubate for 24 hours at 37° C. in a 5% CO2 atmosphere (Thermo Electron Corp., Forma Series II).

US10046005B2. Composition and method of use for combinations of anti-viral protease, polymerase inhibitors and natural bioactive compounds in the treatment of hepatitis C infection (2018-08-14). None [US]. Inventors: Shaker A. Mousa [US].
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