Microwell 96 well optical bottom plate

Manufactured by Thermo Fisher Scientific

The MicroWell 96-well optical bottom plates are a type of laboratory equipment used for cell-based assays and other applications that require optical measurements. The plates feature a 96-well format with an optical-quality bottom, enabling the accurate measurement of absorbance, fluorescence, or luminescence signals from samples within the wells.

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Lab products found in correlation

Axiocam mr r3 camera, by Zeiss (1 mentions) Synergy htx multi mode reader, by Agilent Technologies (1 mentions) Atplite 1step luminescence assay system, by PerkinElmer (1 mentions)

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96-well plates (1169 citations) 96-microwell plates (21 citations) Nunc MicroWell 96-Well Optical-Bottom Plates (11 citations) Microwell plates (9 citations) Nunc™ MicroWell™ 96-Well Optical-Bottom Plates with Polymer Base (7 citations) MicroWell 96 optical bottom plate (2 citations) Nunc™MicroWell™ 96‐Well Optical‐bottom black chimney plates (2 citations) Black Nunc MicroWell 96-well Optical Bottom plates (2 citations)

4 protocols using microwell 96 well optical bottom plate

Quantifying Calcium Transients in 4T1-GCaMP6s Cells

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To detect and quantify calcium transients, 2 × 10⁴ 4T1-GCaMP6s cells/well were plated in Nunc MicroWell 96-well optical bottom plates and incubated at 37 °C for 24 h. 4T1-GCaMP6s cells were infected with VSV-fluc or VSV-p15 at an MOI of 1 and cultured in 10% FBS FluoroBrite DMEM (Thermo Fisher) supplemented with 2mM L-glutamine, 10mM HEPES, 100 µg/mL streptomycin, and 100 units/mL penicillin. 4T1-GCaMP6s were illuminated with a 488 nm laser and images were captured every 0.45 s for 9 min at 20x magnification and 0.8 numerical aperture on a Zeiss Axios Observer Z.1 spinning disk confocal microscope with an AxioCam MR R3 camera. During acquisition, cells were maintained in a humified chamber at 5% CO₂ and 37 °C. The Cellpose cell segmentation algorithm was used to automatically segment GCaMP6s expressing cells from an average intensity projection of the first 100 frames of the timelapse video [44 (link)]. Images were recorded using Zeiss Zen Blue software. Background fluorescence signal was subtracted using the ImageJ rolling ball algorithm [45 (link)], and F/F was calculated by dividing the mean fluorescence intensity of a cell at the time of acquisition by the cell mean intensity over the course of the time series.

Nelson A., McMullen N., Gebremeskel S., De Antueno R., Mackenzie D., Duncan R, & Johnston B. (2024). Fusogenic vesicular stomatitis virus combined with natural killer T cell immunotherapy controls metastatic breast cancer. Breast Cancer Research : BCR, 26, 78.

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Assess Cell Density and Drug Synergy In Vitro

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To assess cell density and synergy of drugs in vitro, 10,000 cells were seeded in black/white Nunc™ Microwell™ 96-well optical bottom plates. After 24 hours, cells were treated with various drug treatments in 20%(v/v) full media and 80%(v/v) EBSS. After 48–72 hours, cells were washed twice with PBS. The ATP-lite 1-step luminescence assay system (Perkin Elmer) was used following manufacturer’s instructions. Luminescence was measured using the Synergy HTX Multi-Mode Reader (Biotek). Synergy was determined using the Loewe Model (ComBenefit software).

Truong A., Yoo J.H., Scherzer M.T., Sanchez J.M., Dale K.J., Kinsey C.G., Richards J.R., Shin D., Ghazi P.C., Onken M.D., Blumer K.J., Odelberg S.J, & McMahon M. (2020). Chloroquine sensitizes GNAQ/11-mutated melanoma to MEK1/2 inhibition. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(23), 6374-6386.

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Laser-Induced Cell Viability Assay

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Fig 2(B) and 2(C) show the overall experimental layout. A Quantel Evergreen 200 (Quantel, USA) was used as the laser beam source. The laser had pulse energy up to 200 mJ and a repetition rate up to 15 Hz with a 532 nm wavelength. A Nunc™ MicroWell™ 96-Well Optical-Bottom Plate (Nunc™ MicroWell™, 165305) with Polymer Base was used as the cell culture dishes. The well plate is shown in Fig 2(C) and 2(E). Cells were pre-cultured in the well plate before conducting the experiment. The well plate was placed on a small aluminum platform fixed on the optical table. A 6 mm diameter hole was drilled in the middle of the aluminum platform to allow for the laser light passage. The hole diameter was determined by the diameter of the laser beam light in the near field. An optical mirror was placed at an angle of 45° under the aluminum platform to re-direct the laser beam vertically. The aluminum L-shape bar shown in Fig 2(C) was used to hold the hydrophone for pressure measurements during the experiment. The L-shape aluminum bar was attached to assembled translation stages with two degrees of freedom–vertically and laser-beam wise. Another degree of freedom was provided by moving the U holder along the slide in the aluminum bar.

Liao Y., Gose J.W., Arruda E.M., Liu A.P., Merajver S.D, & Young Y.L. (2020). Shock wave impact on the viability of MDA-MB-231 cells. PLoS ONE, 15(6), e0234138.

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Endometrial Organoid Encapsulation Optimization

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Four-day-old organoids grown in Matrigel were collected and processed as above to generate single cells. The cell suspension was inspected under the microscope to ensure the presence of dispersed single cells and if needed, filtered through a 30 μm cell strainer to remove cell clumps. Single cells were counted using a hemocytometer then resuspended in the matrix precursor solutions (fPEG-VS) prior to the addition of the crosslinker and Y-27632 (10 μM). In parallel, single cells resuspended in Matrigel served as experimental control during matrix evaluation. In both cases, cells were encapsulated at a density of 500 cells/μL of matrix. Three μL (1,500 cells) of the matrices were loaded into a Nunc MicroWell 96-well optical-Bottom plate and allowed to polymerize for 20 min in a humidified incubator at 37 °C, 95% air, and 5% CO₂. After gelation, 100 μL of OEM or EnOM was loaded into each well. Media was changed every two days. Eight-day old endometrial organoids were used to generate single cells, then a similar encapsulation process in Matrigel or the synthetic ECM was perfomed. The 3 μL droplet size for synthetic ECM (5 μL for passaging experiments) was chosen to conserve resources, facilitate imaging, and reduce inhomogeneities in organoid growth between the rim and center regions for droplets of 15 μL or greater volume.

Hernandez-Gordillo V., Kassis T., Lampejo A., Choi G., Gamboa M.E., Gnecco J.S., Brown A., Breault D.T., Carrier R, & Griffith L.G. (2020). Fully synthetic matrices for in vitro culture of primary human intestinal enteroids and endometrial organoids. Biomaterials, 254, 120125.

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