4 well culture plates

Manufactured by Thermo Fisher Scientific

Sourced in Denmark

The 4-well culture plates are a laboratory equipment designed for cell culture applications. These plates provide a controlled environment for the growth and maintenance of cell lines. Each plate contains four individual wells, allowing for the simultaneous culturing of multiple samples or conditions. The plates are made of durable materials suitable for cell culture use.

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Lab products found in correlation

Tcm 199 medium, by Thermo Fisher Scientific (2 mentions) Vectashield mounting medium, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Anti human hmgb1 antibody, by Cell Signaling Technology (1 mentions) Ultrapure lmp agarose, by Thermo Fisher Scientific (1 mentions) Ibitreat chambers, by Ibidi (1 mentions) Rat tail collagen type 1, by BD (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Las af, by Leica (1 mentions) Confocal microscope, by Leica (1 mentions) Alexa fluor 488 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Phalloidin tetramethylrhodamine b isothiocyanate, by Merck Group (1 mentions) Matrigel, by BD (1 mentions)

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4-well plates (39 citations)

7 protocols using 4 well culture plates

In Vitro Maturation of Sheep Oocytes

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Sheep ovaries were collected from local slaughterhouses and transferred to the laboratory within 1–2 hours. Oocytes were aspirated with 21 G needles in presence of TCM-199 medium (Gibco, Milan, Italy) containing Hepes and 0.005% (w/v) Heparin. Then, all oocytes with an unexpanded cumulus and uniform cytoplasm were in vitro maturated (IVM) in bicarbonate-buffered TCM-199 medium (Gibco Milan, Italy) containing 2 mM glutamine, 0.3 mM sodium pyruvate, 100 μM cysteamine, 10% fetal bovine serum (FBS) (Gibco Milan, Italy), 5 μg/ml FSH (Ovagen, Glenfield, New Zeland), 5 μg/ml LH, and 1 μg/ml estradiol. Maturation was conducted into 4-well culture plates (Nunclon, Roskilde, Denmark) containing 0.4 ml of IVM medium per well and incubated in a humidified atmosphere of 5% CO₂ in air at 39°C for 24 h.

Czernik M., Toschi P., Zacchini F., Iuso D, & Ptak G.E. (2017). Deregulated Expression of Mitochondrial Proteins Mfn2 and Bcnl3L in Placentae from Sheep Somatic Cell Nuclear Transfer (SCNT) Conceptuses. PLoS ONE, 12(1), e0169579.

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Immunofluorescence Analysis of IL-1β and HMGB1

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Primary GECs were cultured in 4-well culture plates (Nunc) containing round glass inserts. Cells were infected at a confluence of 75-80% with either wild-type P. gingivalis or the ndk-deficient mutant strain at an MOI of 100. Six hours later, cells were treated with ATP at a final concentration of 3 mM for an additional 3 h. Cells were washed three times with PBS and fixed with 4% paraformaldehyde for 15 min. Cells were permeabilized using 0.01% Triton X-100 solution in PBS and incubated with the primary antibodies. IL-1β was detected using an FITC-conjugated mouse monoclonal antibody (R&D Systems). HMGB1 was detected using a rabbit polyclonal anti-human HMGB1 antibody (Cell Signaling). P. gingivalis infection was detected using anti-P. gingivalis rabbit polyclonal antibody, coupled with a secondary goat-anti-rabbit polyclonal antibody (Invitrogen), and all inserts were mounted onto glass slides using VectaShield mounting medium (Fisher) containing DAPI stain. The intensity of IL-1β and HMGB1 staining was measured using NIH-ImageJ, and measurements of at least 25 cells, originating from at least three separate replicate experiments, were averaged.

Johnson L., Atanasova K.R., Bui P.Q., Lee J., Hung S.C., Yilmaz Ö, & Ojcius D.M. (2015). Porphyromonas gingivalis attenuates ATP-mediated inflammasome activation and HMGB1 release through expression of a nucleoside-diphosphate kinase. Microbes and infection / Institut Pasteur, 17(5), 369-377.

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Agarose-Based 3D Cell Culture

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Agarose dishes were prepared and cast from 35-well or modified 15-well (cut to size) PDMS moulds (Microtissues) as previously described (Napolitano et al., 2007 (link)) using UltraPure LMP Agarose (Invitrogen) and equilibrated with L-15 supplemented with 1% PSF (Thermo Fisher Scientific) within 4-well culture plates (Nunclon). Cells were seeded in a drop-wise manner as a volume of 75 µl into 35-well dishes or of 35 µl into modified 15-well agarose dishes. Cells were allowed 15-20 min to settle before 750 µl culture medium was added via the medium exchange ports. Culture medium consisted of L-15 supplemented with 10% Embryo Extract (https://zfin.org/zf_info/zfbook/chapt6.html), 3% FBS (Thermo Fisher Scientific), 2% N2 (Thermo Fisher Scientific), 1% PTU (Sigma) and 1% PSF. Cells were incubated at 28.5°C for 48 h before aggregates were harvested for analysis.

Eldred M.K., Charlton-Perkins M., Muresan L, & Harris W.A. (2017). Self-organising aggregates of zebrafish retinal cells for investigating mechanisms of neural lamination. Development (Cambridge, England), 144(6), 1097-1106.

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Culturing Sympathetic Neurons on Collagen

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Sympathetic neurons were plated on collagen coated culture plates. Surface coating was performed under sterile conditions on the same day as the embryo preparation. Rat tail collagen (type I) (BD Bioscience, USA) was diluted using 0.02 N acetic acid to a concentration of 50 µg/ml. Around 200 µl of diluted collagen solution were pipette in every well and left at RT for 1 h. Wells were then washed 4 × 5 min using 500 µl PBS and left under the sterile hood until neurons were plated.
Neurons were plated on either 4-well culture plates (Nunc, Denmark, EU) or 15 µ-Slide 8 well ibiTreat chambers (Ibidi, Germany, EU) at a final concentration of 20,000 cells per 200 or 500 µl (depending on the well-plates used). One day after plating approximately 2/3 of the N2-Medium were substituted by Medium A (N2-Medium without FCS, 1 % B-27 supplement, 0.24 µg/ml 1-β-arabinofuranosylcytosine (Ara-C) and 0.2 % penicillin/streptomycin (Sigma-Aldrich, Germany, EU). On DIV3, all medium was completely substituted by Medium A. Medium was then changed every 2 days until DIV7.

Arnhold M., Dening Y., Chopin M., Arévalo E., Schwarz M., Reichmann H., Gille G., Funk R.H, & Pan-Montojo F. (2016). Changes in the sympathetic innervation of the gut in rotenone treated mice as possible early biomarker for Parkinson’s disease. Clinical Autonomic Research, 26, 211-222.

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Immunofluorescence Imaging of F. nucleatum

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Primary GECs were cultured in 4-well culture plates (Nunc) containing round glass inserts (Warner Instruments). Cells at approximately 80% confluence were infected with F. nucleatum at an MOI of 100 for 1 h. The cells were fixed in 10% neutral buffered formalin and permeabilized with 0.01% Triton X-100 in PBS. F. nucleatum infection was detected using a rabbit anti-F. nucleatum polyclonal antibody (Pacific Immunology) at 1:100 dilution and secondary Alexa Fluor 488-conjugated goat anti-rabbit antibody (Invitrogen) at 1:1000 dilution. Actin was labelled using phalloidin–tetramethylrhodamine B isothiocyanate (Sigma). The immunostained cells were mounted onto glass slides using Vectashield mounting medium (Fisher) containing DAPI stain and examined using a Leica confocal microscope and LAS AF (Leica) software.

Bui F.Q., Johnson L., Roberts J., Hung S.C., Lee J., Atanasova K.R., Huang P.R., Yilmaz Ö, & Ojcius D.M. (2016). Fusobacterium nucleatum infection of gingival epithelial cells leads to NLRP3 inflammasome-dependent secretion of IL-1β and the danger signals ASC and HMGB1. Cellular microbiology, 18(7), 970-981.

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Preparing Hippocampal Neuron-Glia Cultures

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Cell cultures of hippocampal neurons and glial cells were prepared from 2- to 4-day-old mice as described earlier [49 (link)]. In brief, mice were decapitated, the brain was gently but quickly removed and immersed in ice-cold HBSS (Hank's balanced salt solution) containing 20% FCS. Both hippocampi were then isolated, cut with a scalpel into smaller pieces, and trypsinated (5 mg/ml, 10 min, 37°C). The cells were then dissociated by trituration and centrifuged (1500 rpm, 10 min, 4°C), and the pellet obtained was resuspended. Next, this cell suspension was plated on Matrigel (BD Biosciences)-coated glass cover slips, which were placed in 4-well culture plates (Nunc). Dissociated cell cultures were incubated at 37°C (5% CO₂ atmosphere). After 24 h, the medium was replaced with growth medium, and after another 3 days, growth factors as well as half of the medium were refreshed again. Experiments were performed between 3 and 10 days in vitro.

Bebensee D.F., Can K, & Müller M. (2017). Increased Mitochondrial Mass and Cytosolic Redox Imbalance in Hippocampal Astrocytes of a Mouse Model of Rett Syndrome: Subcellular Changes Revealed by Ratiometric Imaging of JC-1 and roGFP1 Fluorescence. Oxidative Medicine and Cellular Longevity, 2017, 3064016.

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Sheep Oocyte Collection and In Vitro Maturation

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Oocyte collection and in vitro maturation was performed as previously described [18 (link)]. Briefly, sheep ovaries were collected from local slaughterhouse (OVIN.COM S.R.L. Pianella, Italy) and transferred to the laboratory within 1–2 hours. Oocytes were aspirated with 21 G needles in presence of TCM-199 medium (Gibco, Thermo Fisher Scientific, Milan, Italy) containing Hepes and Heparin. Then, all oocytes with an unexpanded cumulus and uniform cytoplasm were selected for in vitro maturation (IVM). Maturation was conducted in 4-well culture plates (Nunclon, Roskilde, Denmark) containing 0.4 ml of IVM medium and incubated in a humidified atmosphere of 5% CO₂ in air at 39°C for 24 h.

Arena R., Zacchini F., Toschi P., Palazzese L., Czernik M, & Ptak G.E. (2017). Developmental peculiarities in placentae of ovine uniparental conceptuses. PLoS ONE, 12(11), e0188278.

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