Opticell

Manufactured by Thermo Fisher Scientific

Sourced in United States

The OptiCell is a cell culture chamber designed for live-cell imaging and microscopy. It provides a controlled environment for cell culture and observation. The OptiCell features a gas-permeable, optically clear membrane that allows for efficient gas exchange and optical access for imaging.

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Lab products found in correlation

L glutamine, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Hepes, by Merck Group (2 mentions) Bovine fibrinogen, by Merck Group (1 mentions) Cc 3124, by Lonza (1 mentions) T 75 flasks, by BD (1 mentions) Egm 2, by Lonza (1 mentions) Huvecs, by Lonza (1 mentions) Trypsin, by Lonza (1 mentions) Ethylene diamine tetraacetic acid (edta), by Lonza (1 mentions) Dmem high glucose, by Merck Group (1 mentions) Non essential amino acids (neaa), by Merck Group (1 mentions) Nutrient mixture f 12, by Thermo Fisher Scientific (1 mentions) Atcc ccl 107, by American Type Culture Collection (1 mentions) Poly l lysine (pll), by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions)

Close spelling variants of this product

Nunc® OptiCell™ Cell Culture Systems

7 protocols using opticell

Hydrostatic Pressure Application to Cells

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Application of hydrostatic pressure to cells in vitro was accomplished as previously described (5 (link)–7 (link)) using modifications. Briefly, cells were seeded into Opticell (Thermo Scientific, Rochester, NY) cassettes and allowed to adhere overnight. Pressure was created by connecting the Opticell to an IV bag filled with cell culture media utilizing arterial line tubing. Opticell’s were clamped between perforated stainless steel sheets secured by binder clips. Hydrostatic pressure was modulated by adjusting the height of the IV bag. Pressure was determined using the following formula: P=ρ·h; where P=hydrostatic pressure, ρ=fluid density, and h=the height of the IV bag in relation to the Opticell.
Viability was assessed by pressurizing cells for 24 hours and counting live cells by trypan blue staining. CM was isolated from MLO-Y4 cells by pressurizing cells for 24 hours in RPMI containing 0.1% FBS at 0, 20, and 40 mmHg. CM was prepared as described above. BCA assay was used to quantify protein concentration of cells; no significant (p>0.05) differences were observed after normalization.

Sottnik J.L., Dai J., Zhang H., Campbell B, & Keller E.T. (2015). Tumor-induced pressure in the bone microenvironment causes osteocytes to promote the growth of prostate cancer bone metastases. Cancer research, 75(11), 2151-2158.

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Preparation of Fibrin-Based Hydrogels

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ARSs were prepared using 3, 5, or 10 mg/mL clottable protein by first combining bovine fibrinogen (Sigma-Aldrich), dissolved in degassed (40% O₂ saturation) Dulbecco's modified Eagle's medium (DMEM, Life Technologies), with bovine thrombin (2 U/mL, Thrombin-JMI, King Pharmaceuticals, Bristol, TN, USA), and 1% (v/v) emulsion. The mixture was injected into an OptiCell (Thermo Fisher Scientific Inc., Waltham, MA USA) and allowed to polymerize for 30 min at room temperature. Hydrogels without emulsions were prepared as a sham condition.

Moncion A., Arlotta K.J., Kripfgans O.D., Fowlkes J.B., Carson P.L., Putnam A.J., Franceschi R.T, & Fabiilli M.L. (2015). Design and characterization of fibrin-based acoustically responsive scaffolds for tissue engineering applications. Ultrasound in medicine & biology, 42(1), 257-271.

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Thawing and Culturing Mouse Embryos

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OptiCell (Thermo Fisher Scientific) is a commercially available cell culture device, which comprises a transparent CO₂-permeable membrane and two ports for medium exchange. For thawing, the syringe was removed from the LN₂ (Fig 2e) and then the pipette tip attached to the syringe was inserted into a port of the OptiCell, previously filled with thawing solution. Within 1 min, the HFT with embryos was ejected from the syringe into the warm OptiCell (Fig 2f). Next, a 10-ml syringe containing dilution solution and an empty 10-ml syringe were attached to both ports of the OptiCell. Within 2 min, dilution solution was injected into the OptiCell, and the thawing solution was removed using the empty syringe (Fig 2g). After 3 min, the dilution solution was similarly replaced with washing solution, after 5 min, the washing solution was replaced with CZB medium, and then the OptiCell with the HFT was placed in a CO₂ incubator for 4 days.

Wakayama S., Soejima M., Kikuchi Y., Hayashi E., Ushigome N., Hasegawa A., Mochida K., Suzuki T., Yamazaki C., Shimazu T., Sano H., Umehara M., Matsunari H., Ogura A., Nagashima H, & Wakayama T. (2022). Development of a new device for manipulating frozen mouse 2-cell embryos on the International Space Station. PLoS ONE, 17(10), e0270781.

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Endothelial Cell Culture and Microbubble Interactions

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Human umbilical vein endothelial cells (HUVECs) were cultured in flasks in complete human endothelial growth medium (Lonza CC-3124, Walkersville, MD) in a humidified incubator at 37°C and 5% CO2. HUVEC were plated on the inside of one membrane of an OptiCell^™ (Nunc, Rochester, NY), which consists of two 75 μm thick polystyrene membranes separated by 2 mm, and reached approximately 90% confluency at the time of experiment.
Definity® microbubbles (Lantheus Medical Imaging, Billerca, MA) are C3F8 gas bubbles each encapsulated by a phospholipid shell with a mean diameter in the range of 1.1 – 3.3 μm. After activation following the manufacturer’s protocol, the microbubbles were mixed in complete culture medium at a final concentration of ~ 10⁷ bubble/ml. The culture medium in the OptiCell^™ with HUVECs was replaced by the microbubble mixture right before experiments.

Fan Z., Chen D, & Deng C.X. (2014). Characterization of the dynamic activities of a population of microbubbles driven by pulsed ultrasound exposures in sonoporation. Ultrasound in medicine & biology, 40(6), 1260-1272.

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Culturing Human Umbilical Vein Endothelial Cells

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Human umbilical vein endothelial cells (HUVECs) (Lonza, Verviers, Belgium) were cultured in EGM-2 (Lonza) medium in T75 flasks (BD, Breda, the Netherlands) in a humidified incubator at 37°C with 5% CO₂. Cells were detached with 0.25% Trypsin in EDTA (Lonza) and replated on one side of the acoustically transparent OptiCell^™ (NUNC, Wiesbaden, Germany) chambers. HUVECs were cultured as described before [48 (link)], for two days until 70% confluence to resemble neovasculature endothelial cells.

Skachkov I., Luan Y., van Tiel S.T., van der Steen A.F., de Jong N., Bernsen M.R, & Kooiman K. (2018). SPIO labeling of endothelial cells using ultrasound and targeted microbubbles at diagnostic pressures. PLoS ONE, 13(9), e0204354.

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Culturing Human and Rat Cell Lines for Ultrasound Experiments

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Human melanoma (BLM) cells [20 (link)] were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with Nutrient Mixture F12 (Gibco, Merelbeke, Belgium), supplemented with 10 % fetal bovine serum (FBS) (Hyclone, Thermo Scientific, MA, USA), 20 U/ml penicillin-streptomycin (Gibco), 2 mM l-glutamine (Gibco), and 10 mM HEPES (Sigma-Aldrich®). Human pharynx squamous cell carcinoma (FaDu) cells (ATCC® HTB-43™, LGC Standards GmbH, Wesel, Germany) were cultured in high-glucose DMEM (Sigma-Aldrich®, St. Louis, MO, USA), supplemented with 10 % (v/v) FBS (Sigma-Aldrich®) and 1 % non-essential amino acids (Sigma-Aldrich®). Rat glioma (C6) cells (ATCC® CCL-107™) were maintained in low-glucose DMEM (Sigma-Aldrich®) supplemented with 10 % FBS. Cells were cultured in standard cell culture flasks in a humidified incubator at 5 % CO₂ and 37 °C.
Ultrasound experiments with BLM cells were performed in OptiCells™ (Nunc, Thermo Scientific, MA, USA), wherein 1.3 × 10⁶ cells were plated 1 day prior to the experiment. For ultrasound experiments with FaDu or C6 cells, 1 × 10⁶ cells were seeded into CLINIcell® cell culture chambers (Mabio, Tourcoing, France) 2 days prior to the experiment, to ensure a confluent cell monolayer during the experiment. CLINIcells® were coated with Poly-l-Lysine (Sigma-Aldrich®) before cell seeding for proper cell attachment.

Lammertink B.H., Deckers R., Derieppe M., De Cock I., Lentacker I., Storm G., Moonen C.T, & Bos C. (2017). Dynamic Fluorescence Microscopy of Cellular Uptake of Intercalating Model Drugs by Ultrasound-Activated Microbubbles. Molecular Imaging and Biology, 19(5), 683-693.

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Culturing Human Melanoma Cells

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Human melanoma cells (BLM cells) [26] were grown in culture flasks in a humidified atmosphere with 5% CO 2 at 37 C. The culture medium was Dulbecco's Modified Eagle Medium with Nutrient Mixture F12 (Gibco, Merelbeke, Belgium), supplemented with 10% (v/v) fetal bovine serum (FBS) (Hyclone, Thermo Scientific, MA, USA), 20 U/mL penicillin-streptomycin (Gibco, Merelbeke, Belgium), 2 mM L-glutamine (Gibco, Merelbeke, Belgium) and 10 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) (SigmaeAldrich, Diegem, Belgium). One day before the experiment, cells were harvested by 0.05% trypsin-EDTA (ethylenediaminetetraacetic acid) (Gibco, Merelbeke, Belgium) and replated in Opticells™ (Nunc, Thermo Scientific, MA, USA). For flow cytometry experiments, cells were seeded at a density of 2 Â 10 6 cells/mL, reaching confluency after 1 day. For microscopy experiments, a cell density of 1 Â 10 6 cells/mL was used to obtain a sub-confluent monolayer.

De Cock I., Lajoinie G., Versluis M., De Smedt S.C, & Lentacker I. (2016). Sonoprinting and the importance of microbubble loading for the ultrasound mediated cellular delivery of nanoparticles. Biomaterials, 83.

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