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4 chamber glass bottom dish

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The 4-chamber glass bottom dish is a laboratory equipment designed for cell culture and microscopy applications. It features a transparent glass bottom that allows for high-quality imaging and observation of cells. The dish is divided into four separate chambers, providing a versatile platform for parallel experiments or comparative studies.

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Lab products found in correlation

Fugene reagent, by Promega (1 mentions) Mitored, by Merck Group (1 mentions) Fv1200, by Olympus (1 mentions) Pkh67, by Merck Group (1 mentions) Dmem glutamax, by Merck Group (1 mentions) Ab32575, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Sp8 confocal, by Leica camera (1 mentions)

6 protocols using 4 chamber glass bottom dish

1

Visualizing ROMK2 and Mitochondria in H9c2 Cells

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The H9c2 WT cells were seeded on a 4-chamber glass bottom dish (Cellvis). After overnight incubation, the cells were transiently transfected with plasmid encoding ROMK2 fused with GFP. Transfection was performed using a Fugene reagent (Promega, Madison, WI, USA). Forty-eight hours after transfection, the mitochondria were additionally stained using 100 nM Mitored (Sigma, St. Louis, MO, USA). Confocal images of living cells were acquired using an Olympus FV 1200.
Laskowski M., Augustynek B., Bednarczyk P., Żochowska M., Kalisz J., O’Rourke B., Szewczyk A, & Kulawiak B. (2019). Single-Channel Properties of the ROMK-Pore-Forming Subunit of the Mitochondrial ATP-Sensitive Potassium Channel. International Journal of Molecular Sciences, 20(21), 5323.
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2

Visualizing EV Uptake in MeT-5A Cells

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Purified EVs were incubated with PKH67 (Sigma-Aldrich, USA) for 4 min according to recommendations. The labeling reaction was stopped by 0.5% BSA. The mixture was filtrated through a 0.22 μm filter to remove unbound dye. PKH67-labeled EVs were pelleted and resuspended in PBS. MeT-5A cells (2 × 10^4) were seeded in 4-Chamber Glass Bottom Dish (Cellvis, USA), after adherence, 10 μg PKH67-labeled EVs were incubated with MeT-5A cells for 24 h. After being fixed with 4% paraformaldehyde for 10 min, cell skeleton was labeled with TRITC Phalloidin, nuclei were stained with DAPI and the uptake of EVs into MeT-5A cells was visualized by confocal laser scanning microscope.
Wang C., Wang J., Shen X., Li M., Yue Y., Cheng X., Lu W., Wang X, & Xie X. (2021). LncRNA SPOCD1-AS from ovarian cancer extracellular vesicles remodels mesothelial cells to promote peritoneal metastasis via interacting with G3BP1. Journal of Experimental & Clinical Cancer Research : CR, 40, 101.
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3

Transfection and Live-Cell Imaging of U-2 OS Cells

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U-2 OS cells (ATCC, HTB-96) were maintained in DMEM + GlutaMAX (Sigma-Aldrich, 61965-026) supplemented with 10% FBS at 37°C in 5% CO2. Two days before imaging, the cells were plated onto 4-chamber Glass Bottom Dish (Cellvis, D35C4-20-1.5-N) to achieve 30% confluency. On the following day, the cells were transfected with the mammalian expression vector pcDNA4.0/TO_EGFP-tau or pEGFP-C1_MAP4, or co-transfected with the pcDNA4.0/TO_EGFP-tau and pmScarlet-i alphaTubulin-C1 expression vectors using the HP-Xtreme transfection reagent (Sigma-Aldrich, 636624400). The pmScarlet-i_alphaTubulin_C1 was a gift from Dorus Gadella (Addgene plasmid #85047; http://n2t.net/addgene:85047; RRID:Addgene_85047). The cells were imaged 24h after transfection, with DMEM medium exchanged for FluoroBright DMEM medium (Sigma-Aldrich, A1896701).
Siahaan V., Tan R., Humhalova T., Libusova L., Lacey S.E., Tan T., Dacy M., Ori-McKenney K.M., McKenney R.J., Braun M, & Lansky Z. (2022). Microtubule Lattice Spacing Governs Cohesive Envelope Formation of Tau Family Proteins. Nature chemical biology, 18(11), 1224-1235.
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4

Immunofluorescence Analysis of Fibroblast Activation

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The fibroblasts were seeded in a 4‐chamber glass‐bottom dish (Cellvis) and then treated with TGF‐β1 for 48 hours. After fixing with carbinol18 and blocking with 5% BSA, the fibroblasts were incubated with the primary antibodies (ɑ‐SMA, Abcam, ab32575, 1:500) at 4°C overnight and with Cy3‐conjugated goat anti‐rabbit secondary antibodies (1:200, Beyotime Bio, Shanghai, China) at room temperature for 1 hour. The nucleus was labelled with DAPI (Beyotime Bio, Shanghai, China) for 10 minutes. The fibroblasts were imaged with a fluorescence microscope (Olympus, Tokyo, Japan).
Wu Q., Han L., Gui W., Wang F., Yan W, & Jiang H. (2020). MiR‐503 suppresses fibroblast activation and myofibroblast differentiation by targeting VEGFA and FGFR1 in silica‐induced pulmonary fibrosis. Journal of Cellular and Molecular Medicine, 24(24), 14339-14348.
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5

Immunofluorescence Staining of MeT-5A Cells

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MeT-5A cells (2 × 10^4) were seeded in 4-Chamber Glass Bottom Dish (Cellvis, USA), after treatment, cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 5 min and blocked with 2% BSA for 30 min. Then cells were incubated with primary antibodies at 4 °C overnight and secondary antibody at room temperature for 1 h and stained with DAPI. A confocal laser scanning microscope was used to observe and take images. Primary and secondary antibodies used were listed in Table S2.
Wang C., Wang J., Shen X., Li M., Yue Y., Cheng X., Lu W., Wang X, & Xie X. (2021). LncRNA SPOCD1-AS from ovarian cancer extracellular vesicles remodels mesothelial cells to promote peritoneal metastasis via interacting with G3BP1. Journal of Experimental & Clinical Cancer Research : CR, 40, 101.
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6

Optically Induced Droplet Formation

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The optoDroplet assay was adapted from previous study (Shin et al., 2017) . Plasmids were co-transfected with PEI transfection reagent in 293T cells. For transfection, 293T cell were plated 24 h prior to transduction in 4-chamber glass bottom dish (Cellvis). After transfection, the dishes were cultured in 37 C incubator for 20-24 h. Imaging was performed on Leica SP8 confocal. For global activation and imaging, cells were imaged typically by use of two laser wavelengths. Droplet formation was induced by 488nm light for Cry2 activation with the increasing activation intervals of 10 s, 20 s, 30 s, 40 s, 50 s and 60 s. mCherry imaging was stimulated with 568nm light.
Liu X., Shen J., Xie L., Wei Z., Wong C., Li Y., Zheng X., Li P, & Song Y. (2020). Mitotic Implantation of the Transcription Factor Prospero via Phase Separation Drives Terminal Neuronal Differentiation. Developmental cell, 52(3).
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