8 well glass bottom chambers

Manufactured by Ibidi

Sourced in Germany

The 8-well glass-bottom chambers are a type of cell culture accessory designed for microscopy applications. They provide a convenient platform for culturing cells and observing them under a microscope. Each chamber features a glass bottom that allows for high-quality optical imaging and analysis.

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Lab products found in correlation

Las x sp8, by Leica (1 mentions) Concanavalin a (cona), by Merck Group (1 mentions) Tcs sp8 microscope, by Leica (1 mentions) Lsm 710 confocal microscope, by Zeiss (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Perfect focus, by Nikon (1 mentions) Eclipse ti e inverted microscope, by Nikon (1 mentions) L 15 medium, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Hepes ph 7, by Merck Group (1 mentions) Atto647n dppe, by ATTO-TEC (1 mentions)

Close spelling variants of this product

8-well chambers (44 citations) Ibidi glass-bottom chambers (5 citations) 8-well glass chambers (2 citations)

5 protocols using 8 well glass bottom chambers

Fluorescence Microscopy Protocols for Yeast

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For fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments, overnight yeast pre-cultures were re-inoculated into appropriate synthetic media supplemented with 2× access of adenine and incubated at 30°C for 12 h before imaging. Cells were imaged in glass bottom 8-well chambers (IBIDI) pre-coated with ConA (Sigma-Aldrich).
All FRAP and FLIP experiments were performed with a Leica TCS SP8 microscope (Leica DMI6000B-CS) using 63× 1.4NA Oil HC PL APO CS2 objective. The microscope was equipped with a Laser unit for confocal acquisition (AOBS system): 458, 477, 488, 496, 514nm lines Argon laser; 405nm, 561nm, 633nm lasers, and controlled by Leica LAS X SP8 Version 1.0 software.

Onischenko E., Tang J.H., Andersen K.R., Knockenhauer K.E., Vallotton P., Derrer C.P., Kralt A., Mugler C.F., Chan L.Y., Schwartz T.U, & Weis K. (2017). Natively unfolded FG-repeats stabilize the structure of the nuclear pore complex. Cell, 171(4), 904-917.e19.

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Colocalization and Signaling Assays of KRAS4B

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HEK293T cells were cultured in glass-bottom 8-well chambers (ibidi GmbH) for colocalization assays or in a 12-well plate for signaling assays for 1 day prior to transfection. On the day of transfection at 70–90% confluency, media were replaced with antibiotics-free complete media (DMEM supplemented with 10% FBS). The vectors encoding mCherry-fused monobodies were constructed by cloning their genes amplified using primers, NheI_mcherrF mcherr_flagR, Flag_Mb_F, and mb_ApAI_R (Supplementary Table 4), into the pEGFP vector at the NheI and ApAI restriction enzyme sites. The pEGFP KRAS4B vector was a gift of Prof. Mark Philips. Transfection of pEGFP vectors encoding the appropriate mCherry fused monobodies and EGFP fused KRAS4B constructs was performed with lipofectamine 3000 (Thermo Fisher Scientific) and according to the manufacturer’s recommended protocol. On the following day, transfected cells were imaged with a LSM710 confocal microscope (Zeiss) for colocalization experiment or harvested for western blot analysis.

Teng K.W., Tsai S.T., Hattori T., Fedele C., Koide A., Yang C., Hou X., Zhang Y., Neel B.G., O’Bryan J.P, & Koide S. (2021). Selective and noncovalent targeting of RAS mutants for inhibition and degradation. Nature Communications, 12, 2656.

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SARS-CoV-2 Infection of Engineered Organoids

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8-well glass-bottom chambers (ibidi, Graefling, Germany) were coated with 2.5% human collagen in water for 1 h prior to organoid seeding. Organoids were collected at a ratio of 100 organoids per well of a 48-well plate. Collected organoids were spun at 450 × g for 5 min, and the supernatant was removed. Organoids were washed with cold 1× PBS and spun at 450 × g for 5 min. PBS was removed, and organoids were digested with 0.05% Trypsin-EDTA (Life Technologies) for 5 min at 37°C. Digestion was stopped by addition of serum-containing medium. Organoids were spun at 450 × g for 5 min, and the supernatant was removed before organoids were resuspended in normal growth medium at a ratio of 250 μL medium per well. The collagen mixture was removed from the ibidi chambers, and 250 μL of organoids were added to each well. Forty-eight hours after seeding, the medium was removed and replaced with differentiation medium for 4 days. Following organoid differentiation, organoids were transduced with AAV vectors at an MOI of 10⁵. Three days after transduction, the medium was removed, and transduced organoids were infected with SARS-CoV-2 (BavPat1, passage 3) at an MOI of 1 for 1 h. Following infection, virus was removed, cells were washed with 1× PBS, and differentiation medium was replaced. Twenty-four hours after infection, supernatants and RNA were harvested.

Becker J., Stanifer M.L., Leist S.R., Stolp B., Maiakovska O., West A., Wiedtke E., Börner K., Ghanem A., Ambiel I., Tse L.V., Fackler O.T., Baric R.S., Boulant S, & Grimm D. (2022). Ex vivo and in vivo suppression of SARS-CoV-2 with combinatorial AAV/RNAi expression vectors. Molecular Therapy, 30(5), 2005-2023.

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Visualizing Microtubule Polarity in RPE-1 Cells

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To examine microtubule polarity, RPE‐1 cells expressing EB3‐GFP (−DOX, +DOX, H₂O₂, Tubacin) or EB3‐tdTomato (αTAT1 OE) were seeded onto 8‐well glass bottom chambers (iBidi) overnight at low confluency (3 × 10⁴ per well). The following day, H₂O₂ and Tubacin conditions were treated as previously described before live imaging using an Eclipse Ti‐E inverted microscope (Nikon) equipped with a CSU‐X1 Zyla 4.2 camera (Ti‐E, Zyla; Andor), including a Yokogawa Spinning Disk, a precision motorized stage, and Nikon Perfect Focus, all controlled by NIS‐Elements Software (Nikon). The microscope was enclosed within temperature‐ and CO₂‐controlled environments that maintained an atmosphere of 37°C and 5% humidified CO₂ for live‐cell imaging. 60× 1.45‐NA oil objective was used to capture images every second for 30 s. EB3‐positive comets were tracked using the TrackMate plugin for Fiji (Tinevez et al, 2017 (link)), using automatic detection and filtering for tracks longer than 7 s to calculate the total number of EB3 comets. Identification of reverse polarity microtubules, defined as an EB3 comet traveling towards the center of the cell, was then performed manually. Temporal projections were made using the “Temporal‐Color Code” feature in Fiji and colored using the Turbo LUT.

Monteiro P., Yeon B., Wallis S.S, & Godinho S.A. (2023). Centrosome amplification fine tunes tubulin acetylation to differentially control intracellular organization. The EMBO Journal, 42(16), e112812.

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Jurkat T-cell Labeling and Measurement

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Jurkat T-cells were cultured in RPMI-1640 (Sigma Aldrich, UK) media supplemented with 10% FBS (Sigma Aldrich), 2 mM l-Glutamine (Sigma Aldrich), 100 U/mL Penicillin (Sigma Aldrich), 0.1 mg/mL Streptomycin (Sigma Aldrich) and 10 mM HEPES pH 7.4 (Sigma Aldrich). 1 million cells were spun down for 5 min at 2000 rpm and washed with 1 mL of L15 medium (Life Technologies). After spinning down again the cells were labelled by resuspending in L15 medium containing 0.4 μg/mL Atto647N-DPPE (AttoTec). The cells were labelled at 37 °C shaking at 300 rpm for 15 min. After washing with L15, the cells were resuspended and kept in L15 for not longer than 1 h on room temperature. Measurements were performed in 8-well glass-bottom chambers (Ibidi). Prior to the measurements the glass was coated with PLL using a 0.01% PLL-solution (Poly-l-Lysine) (Sigma Aldrich) for 1 h at room temperature and washed three times with L15.

Waithe D., Schneider F., Chojnacki J., Clausen M.P., Shrestha D., de la Serna J.B, & Eggeling C. (2018). Optimized processing and analysis of conventional confocal microscopy generated scanning FCS data. Methods (San Diego, Calif.), 140-141, 62-73.

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