Confocal dishes

Manufactured by MatTek

Sourced in United States

Confocal dishes are specialized cell culture vessels designed for use with confocal microscopy. They feature a thin, optically-clear glass bottom that allows for high-resolution imaging of cells and tissues during live-cell experiments.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using confocal dishes

Characterization of Murine Induced Pluripotent Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

miPSC were plated in confocal dishes (MatTek) for immunofluorescence analysis 48 h after plating using the miPSC Characterization kit (Applied Stemcell). Briefly, cells were fixed, permeabilized, and stained overnight at 4 °C with the primary antibodies for Sox2, SSEA-1 and Oct4. After several washes the cells were incubated with a secondary antibody and DNA staining solution. Alkaline phosphatase activity assay was performed (Applied Stemcell). Stained cells were imaged using a fluorescent microscope.
For RT–PCR, RNA was extracted using the RNeasy Plus Mini Kit (Qiagen). Genomic DNA contamination was removed using the gDNA spin column. cDNA was generated using Applied Biosystems High-Capacity cDNA Reverse Transcription kit. Gene-specific primers of the miPSC Characterization Kit (Applied Stemcell) were used to amplify target sequences. Actin was used as housekeeping gene, which encodes a cellular cytoskeleton protein. PCR reactions were performed on Mastercycler nexus (Eppendorf) and visualized on 2% agarose gels.

Deuse T., Hu X., Gravina A., Wang D., Tediashvili G., De C., Thayer W.O., Wahl A., Garcia J.V., Reichenspurner H., Davis M.M., Lanier L.L, & Schrepfer S. (2019). Hypoimmunogenic derivatives of induced pluripotent stem cells evade immune rejection in fully immunocompetent allogeneic recipients. Nature biotechnology, 37(3), 252-258.

+ Open protocol

+ Expand

Assessing Gap Junction Function in MSCs Under Smoke Exposure

Check if the same lab product or an alternative is used in the 5 most similar protocols

Functionality of gap junction-mediated intercellular communication upon smoke exposure was assessed using the FRAP assay. MSCs were seeded onto confocal dishes (MatTek Corporation, Massachusetts, USA) and treated with cigarette or e-cigarette extracts. On day 14, cells were labelled with calcein-AM (C3100, Molecular Probes, New York, USA), by incubation in 1 μM calcein-AM reconstituted in DMSO, for one hour at 37 °C. Experiments were performed by live imaging at 37 °C using the LSM 710 (63x/1.46 Oil Plan-Apochromatic objective). Experimental cells or regions of interest (ROI) were chosen based on cell confluence, cell-cell contact (allowing gap junction communication) and comparable fluorescence intensity of neighbouring cells. Selected cells were photo-bleached using the 408 nm laser at 10% laser power. Fifteen iterations at 10-second intervals achieved a 50% decrease of ROI fluorescence intensity. Fluorescence recovery was assessed under the 488 nm laser. Images were captured at 10-second intervals and fluorescence intensity of the ROI was quantified over time and normalized to that of control, unbleached, calcein-loaded cells.

Shaito A., Saliba J., Husari A., El-Harakeh M., Chhouri H., Hashem Y., Shihadeh A, & El-Sabban M. (2017). Electronic Cigarette Smoke Impairs Normal Mesenchymal Stem Cell Differentiation. Scientific Reports, 7, 14281.

+ Open protocol

+ Expand

Immunofluorescence Staining of Cellular Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were plated and grown on confocal dishes (MatTek, Ashland, MA) in standard media to appropriate (60–80%) confluence. Cells were then washed three times for 3 min with 1% PBS and fixated in 4% paraformaldehyde (Sigma) for 10 min. Cells were washed three times in 1% PBS for 3 min and permeabilized in 100% methanol for 15 min. Cells were washed in 1% PBS three times for 3 min each and then blocked using 5% bovine serum albumin (BSA, Sigma) for 60 min. Cells were then washed in 1% TBS-T three times for 3 min and incubated in primary antibodies [rabbit anti-Cav-1–1:100 (Abcam), mouse anti-Cav-1–1:100 (BD Technologies), rabbit anti-MnSOD- 1:100 (Abcam), goat anti-MnSOD-1:100 (Santa Cruz), rabbit anti-Nrf2–1:100 (Santa Cruz)] in 1% BSA in 1% TBS-T overnight at 4°C in a humid chamber. Cells were washed three times in 1% TBS-T for 3 min and incubated with secondary fluorescent antibodies [rabbit Alexfluor-568, mouse Alexafluor-488 and goat Alexafluor-647 (Thermo Fisher Scientific)] for 2 h at room temperature in a dark, humid chamber. Cells were then washed three times in 1% TBS-T for 3 min and incubated with 50 μM DAPI (Thermo Fisher Scientific) for 30 min. Cells were washed twice in 1% TBS-T for 3 min and once in PBS for 5 min and then imaged using LSM-510META (Carl Zeiss Microscopy, Thornwood, NY).

Hart P.C., Ratti B.A., Mao M., Ansenberger-Fricano K., Shajahan-Haq A.N., Tyner A.L., Minshall R.D, & Bonini M.G. (2015). Caveolin-1 regulates cancer cell metabolism via scavenging Nrf2 and suppressing MnSOD-driven glycolysis. Oncotarget, 7(1), 308-322.

+ Open protocol

+ Expand

Intracellular Localization of TLR7 Signaling Components in BMDCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

BMDCs were seeded in confocal dishes (Mattek) at a final concentration of 1×10⁵ cells/mL in RPMI-complete media. The following day, cells were treated with 10 μg/mL GRD for 5 minutes and 15 minutes at 37C. Cells were carefully washed with PBS and fixed with 2% PFA for 10 minutes at room temperature in the dark. Fixed cells were permeabilized with SAP buffer (2% saponin in PBS) for 1 hour at 37C. Following permeabilization, BMDCs were intracellularly stained with antibodies to mouse TLR7, MyD88, EEA1, or LAMP1 (Thermo Scientific), followed by staining with secondary antibodies fluorescently labeled with AlexaFluor488 or AlexFluor550 (Invitrogen). Cells were washed 3 times with PBS. Finally, DNA was stained with 1:1000 dilution of DAPI (Invitrogen) for 10 minutes at RT. Cells were visualized using a Nikon confocal microscope and analysis performed using ZEN software. Data analysis and processing of images was conducted using ImageJ software (NIH).

Ramirez-Ortiz Z.G., Prasad A., Griffith J.W., Pendergraft WF I.I.I., Cowley G.S., Root D.E., Tai M., Luster A.D., Khoury J.E., Hacohen N, & Means T.K. (2015). TREML4 amplifies TLR7-mediated signaling during antiviral responses and autoimmunity. Nature immunology, 16(5), 495-504.

+ Open protocol

+ Expand

Intracellular Localization of TLR7 Signaling Components in BMDCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Cardiomyocyte Isolation and Culture

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

On day (7 + 4), mESC-CMs were isolated as previously described [19 (link)–21 (link)]. The isolated mESC-CMs were plated on cover glass (Thermo Fisher Scientific) in a 24-well plate or confocal dishes (MatTek, Ashland, MA, USA) that have been pre-coated with 20 μg/mL laminin (Thermo Fisher Scientific) in 0.1% gelatin in 37 °C, 5% CO₂ incubator.
TRPA1 activator or blocker treatment experiments were performed on day (7 + 5). For experiments involving TRPA1 knockdown or TRPA1 knockdown concomitant with PGC-1α overexpression, mESC-CMs were infected with adenoviruses on day (7 + 5) and the experiments were performed on day (7 + 9). For TRPA1 or PGC-1α overexpression experiments, mESC-CMs were infected with adenoviruses on day (7 + 5) and the experiments were performed on day (7 + 7).

Ding Q., Liu X., Qi Y., Yao X, & Tsang S.Y. (2023). TRPA1 promotes the maturation of embryonic stem cell-derived cardiomyocytes by regulating mitochondrial biogenesis and dynamics. Stem Cell Research & Therapy, 14, 158.

+ Open protocol

+ Expand

Ethidium Bromide Uptake Assay for PANX1 Activity

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

MDA-MB-231 and MDA-PANX1^– cells were seeded onto glass-bottom dishes (confocal dishes, MatTek Corporation, Ashland, MA, USA), at a seeding density of 1.2 × 10⁴ cells per dish. At 60–70% confluence, cells were washed twice with 200 µL of low divalent ion extracellular solution (145 mM NaCl, 5 mM KCl, 13 mM glucose, 0.2 mM CaCl₂, 10 mM Hepes, pH 7.3). MDA-MB-231 cells were then treated with 1 mM PBN for 15 m at 37 °C, before applying low divalent ion extracellular solution containing 25 µM ethidium bromide ([EtBr]; MP Biomedicals, Germany) [99 (link)]. Measurement of EtBr uptake by the cells was performed by recording EtBr fluorescence by live imaging for 15 m at 37 °C using a Laser Scanning Confocal Microscope (LSM710, Carl Zeiss, Oberkochen, Germany). The assay was also performed in normal divalent solution (145 mM NaCl, 5 mM KCl, 2 mM CaCl₂, 1 mM MgCl₂, 13 mM glucose, 10 mM HEPES, pH 7.3) and at room temperature, with the addition of 1 mM ATP to stimulate PANX1 channel opening, as previously documented [49 (link)]. Results from this assay are displayed in Figure S2.

Jalaleddine N., El-Hajjar L., Dakik H., Shaito A., Saliba J., Safi R., Zibara K, & El-Sabban M. (2019). Pannexin1 Is Associated with Enhanced Epithelial-To-Mesenchymal Transition in Human Patient Breast Cancer Tissues and in Breast Cancer Cell Lines. Cancers, 11(12), 1967.

+ Open protocol

+ Expand

Comprehensive Characterization of hiPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

hiPSCs were plated in confocal dishes (MatTek) for immunofluorescence analysis 48 h after plating using the hiPSC Characterization Kit (Applied Stem Cell). Briefly, cells were fixed, permeabilized and stained overnight at 4 °C with the primary antibodies for OCT4, SOX2, SSEA4, TRA-1–60 and TRA-1–81 (Applied Stem Cell). After several washes, the cells were incubated with a secondary antibody and DNA staining solution. Alkaline phosphatase activity assay was performed (Applied Stem Cell). Stained cells were imaged using a fluorescent microscope.
For RT–PCR, RNA was extracted using the RNeasy Plus Mini Kit (Qiagen). Genomic DNA contamination was removed using the gDNA spin column. cDNA was generated using Applied Biosystems High-Capacity cDNA Reverse Transcription Kit. Gene-specific primers of the hiPSC Characterization Kit (Applied Stem Cell) were used to amplify target sequences Actin was used as housekeeping gene. PCR reactions were performed on Mastercycler nexus (Eppendorf) and visualized on 2% agarose gels.

+ Open protocol

+ Expand

Tracking Pos3Aa-p53 Protein Uptake

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified Pos3Aa-p53 crystals were labeled with Alexa Fluor 568 C5 Maleimide (Alexa568-Pos3Aa-p53). 4T1 cells were seeded at 35,000 cells/well in confocal dishes (MatTek) and incubated overnight at 37°C in a 5% CO₂ incubator for cell attachment. On the next day, the cells were washed with 1× PBS, and then treated with 200 nM Pos3Aa-p53 crystals for 24, 48 or 72 hours. At the end of each incubation period, cells were washed three times with 20 U/mL heparin in PBS and then stained with 0.2 µg/mL Hoechst 33 342 (Life Technologies) and 50 nM LysoTracker Green DND-26 (Thermo Scientific) at 37°C for 10 min prior to imaging on a Leica SP8 confocal microscope.

Yang Z., Sun J.K., Lee M.M, & Chan M.K. (2022). Restoration of p53 activity via intracellular protein delivery sensitizes triple negative breast cancer to anti-PD-1 immunotherapy. Journal for Immunotherapy of Cancer, 10(9), e005068.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!