Compact digital microplate shaker

Manufactured by Thermo Fisher Scientific

The Compact Digital Microplate Shaker is a laboratory instrument designed to gently mix and agitate samples in microplates. It features a compact and portable design, providing consistent and controlled shaking motion for a wide range of applications.

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Lab products found in correlation

V plex, by Mesoscale (2 mentions) V plex, by Agilent Technologies (2 mentions) Elx5012, by Agilent Technologies (2 mentions) Meso scale discovery msd, by Mesoscale (1 mentions) Mops ez rich defined medium, by Teknova (1 mentions) Read buffer, by Mesoscale (1 mentions) Breathe easy sealing membrane, by Merck Group (1 mentions) M200 pro microplate reader, by Tecan (1 mentions)

6 protocols using compact digital microplate shaker

Multiplex Cytokine Profiling of Stimulated T-cells

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Interferon-γ (IFN-γ), IL-2 and tumor necrosis factor-α (TNF-α) concentrations were measured using multiplex cytokine arrays [V-PLEX, Meso Scale Discovery (MSD), Meso Scale Diagnostics, Rockville, MD] according to the manufacturer’s instructions. Assays were performed by the Human Immune Monitoring Shared Resource, University of Colorado School of Medicine, Aurora, CO. Briefly, thawed and washed T-cells were plated at 2 × 10⁵ viable cells/well in 96-U well plates and incubated for 6 h in a 37 °C incubator with 5% CO₂ prior to addition of Dynabeads (anti-CD3/anti-CD28; 2 µL/well). Cells were stimulated with beads for 18 h, then supernatants were harvested and stored at − 80 °C until assay. All assays included the T-cell comparator control described above for flow cytometry assays.
Pre-coated V-PLEX plates (MSD) were washed using an automated plate washer (BioTek ELX5012), and calibrators or thawed, diluted supernatants were added, followed by further incubation for 2 h at RT on a Compact Digital Microplate shaker (Thermo Fisher Scientific) at 600 rpm. Plates were again washed, and detection antibodies were added and incubated for 2 h at RT. After washing, 2× Read Buffer (MSD) was added and plates were immediately read on a MesoQuickPlex SQ120 electrochemiluminescent plate reader. Data were analyzed using Workbench software (MSD).

Coeshott C., Vang B., Jones M, & Nankervis B. (2019). Large-scale expansion and characterization of CD3+ T-cells in the Quantum® Cell Expansion System. Journal of Translational Medicine, 17, 258.

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Bacterial Cloning and Fluorescence Assay

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The culture medium lysogeny broth (LB) was used for cloning purposes. Transformed cultures were plated out on lysogenic broth agar (LBA) and grown overnight at 30 °C. If required, LB and LBA medium were supplemented with appropriate antibiotics (100 μg/mL ampicillin or 50 μg/ml kanamycin). For microscopy and the indirect analysis method, strains were grown in minimal medium glucose (MMGlc). If using DH10B cells, 0.4% casamino acids were added. If required, the culture medium was supplemented with appropriate antibiotics and inducers (0–1% L-arabinose or 0–0.2 mM IPTG). For fluorescence experiments with the smart library of sRNAs, strains were grown in MOPS EZ Rich Defined medium (Teknova) with 100 μg/mL ampicillin and 2% glucose as carbon source. For all fluorescent measurements, independent of medium (EZ Rich or MMGlc) used, precultures were grown overnight on a Compact Digital Microplate Shaker (Thermo Fisher Scientific) at 30 °C and 800 rpm, in 96-well flat bottomed microtiter plates (Greiner Bio-One), in 150 μL LB per well with appropriate antibiotics, enclosed by a Breathe-Easy sticker. Precultures were used for 1/150 dilution in 150 μL of defined medium (EZ Rich or MMGlc) containing a specific concentration of the inducer, and were grown for 24 h at 30 °C and 800 rpm unless stated otherwise.

Guidi C., De Wannemaeker L., De Baets J., Demeester W., Maertens J., De Paepe B, & De Mey M. (2022). Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli. Microbial Cell Factories, 21, 260.

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CD19 CAR T-cell Cytokine Profiling

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CD19 CAR T cells were quick-thawed in a 37°C water bath, washed in complete media, counted, and resuspended in complete media. A total of 7.5×10⁵ CD19 CAR T cells were plated in a 96-well round bottom plate with ±2.5×10⁵ Raji cells and incubated for 18 hours in a 37°C incubator with 5% CO₂. The supernatants were harvested after spinning the plate at 500× g for 10 minutes and stored at −80°C. A multiplex cytokine array (V-PLEX; MesoScale Discovery, Rockville, MA, USA) was used to measure cytokines in the supernatants according to the manufacturer’s instructions. Briefly, supernatants were thawed, spun at 2,000× g for 3 minutes, and diluted 1:1 in assay diluent to measure IL-10, IL-12p40, IL-13, IL-1β, IL-4, and IL-6 and diluted 1:100 to measure IL-2, IL-8, IFN-γ, and TNF-α. Pre-coated V-PLEX plates were washed using an automated plate washer (BioTek ELX5012), 50 µL of calibrators or diluted supernatants were added, and plates were incubated for 2 hours at room temperature on a Compact Digital Microplate shaker (Thermo Fisher Scientific) at 600 rpm. Plates were washed, and 25 µL of diluted detection antibodies was added and incubated for 2 hours at room temperature. After washing, 2× Read Buffer (MesoScale Discovery) was added, and the plates were immediately read on a MesoQuickPlex SQ120 electrochemiluminescence plate reader (MSD).

Zhang W., Jordan K.R., Schulte B, & Purev E. (2018). Characterization of clinical grade CD19 chimeric antigen receptor T cells produced using automated CliniMACS Prodigy system. Drug Design, Development and Therapy, 12, 3343-3356.

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Biomaterial-Free Cardiac Patch Bioprinting

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We have previously described and published our protocol for 3D bioprinting of monolayer cardiac patches of size ~ 3.6 × 3.6 mm using only cardiospheres, without the use of biomaterials(Ong, Fukunishi, Zhang, et al. 2017 (link)). The thickness of the mono layer patch in this study was about 350–400 um. In brief, a 3D bioprinter (Regenova, Cyfuse Biomedical K.K., Tokyo, Japan) was used to create biomaterial-free cardiac patches. The predetermined 3D geometry (one single layer of spheroids) was designed using the software of the 3D bioprinter. In a sterile environment, the 3D bioprinter identified the locations of the spheroids within the wells of the ultra-low adhesion 96-well tissue culture plate by automated software detection, a robotic arm then used vacuum suction to pick up, transfer and load the cell aggregates individually onto a needle array in exact spatial coordinates according to the 3D design. After bioprinting, the cardiac patch was allowed to mature for 3 days (72 hours) culture within the needle array, while placed on a shaker (Compact Digital Microplate Shaker, Thermo Scientific, Waltham, MA) at 150 rounds per minute (rpm) in the incubator. During this time, the spheroids fuse to form a cardiac patch with synchronous beating function. The needle array was removed (decannulation) and the cardiac patches were cultured for 7–10 days before implantation surgery.

Yeung E., Fukunishi T., Bai Y., Bedja D., Pitaktong I., Mattson G., Jeyaram A., Lui C., Ong C.S., Inoue T., Matsushita H., Abdollahi S., Jay S.M, & Hibino N. (2019). Cardiac Regeneration Using Human iPSC-derived Biomaterial-free 3D bioprinted Cardiac Patch in vivo. Journal of tissue engineering and regenerative medicine, 13(11), 2031-2039.

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In vivo Translational Inhibition Assay

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For in vivo assessment of translational inhibition, strains were plated on LBA plates containing 100 μg ml^-1 ampicillin and 50 μg ml^-1 kanamycin. After overnight incubation, three colonies were inoculated in 150 μl MOPS EZ rich medium, covered by a Breathe-Easy sealing membrane (Sigma-Aldrich), and grown overnight on a Compact Digital Microplate Shaker (Thermo Scientific) at 800 rpm and 37°C. Subsequently, these cultures were 1:100 diluted in 150 μl of fresh MOPS EZ rich medium and grown on a Compact Digital Microplate Shaker until late log phase (6 h) at 800 rpm and 37°C. Subsequently, fluorescence and OD were measured using a Tecan M200 pro microplate reader. Precultures were grown in Greiner bio-one (Vilvoorde, Belgium) polystyrene F-bottom 96 well plates. Fluorescence and OD measurements were performed after growth in Greiner bio-one (Vilvoorde, Belgium) black μclear 96 well plates. For measuring mKate2 expression an excitation wavelength and an emission wavelength of 588 nm and 633 nm were used, respectively. OD was measured at a wavelength of 700 nm to reduce bias in estimates of cell abundance [44 (link)].

Peters G., Maertens J., Lammertyn J, & De Mey M. (2018). Exploring of the feature space of de novo developed post-transcriptional riboregulators. PLoS Computational Biology, 14(8), e1006170.

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Th and 225Ac Adsorption on HCST

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Experimental solutions, consisting of 20 mg L⁻¹ of Th and 3.7 × 10⁴ Bq ²²⁵Ac in 0.5 M sodium acetate (NaOAc), were adjusted to a pH of 1, 2, 3, 4 or 5 with 10 M NaOH or 69% HNO₃. Inorganic ion-exchangers HCST (100 ± 0.5 mg) were added to 10 mL of the metal containing solution. The tubes were shaken (using Thermo Scientific compact digital microplate shaker) for 12 hours at room temperature. The tubes were centrifuged (4000 rpm, 2598 RCF) for 4 minutes and the aqueous phases were separated using 0.2 µm micro syringe filter. An aliquot from the aqueous phases was diluted in 2% nitric acid and analysis performed.

Fitzsimmons J., Abraham A., Catalano D., Younes A., Cutler C.S, & Medvedev D. (2019). The application of poorly crystalline silicotitanate in production of 225Ac. Scientific Reports, 9, 11808.

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