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Cellcarrier ultra plates

Manufactured by PerkinElmer
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CellCarrier Ultra plates are high-quality cell culture plates designed for a variety of applications. These plates feature a polystyrene surface that has been treated to enhance cell attachment and growth. The plates are available in a range of well formats and are suitable for use in cell-based assays, drug screening, and other cell culture experiments.

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Lab products found in correlation

Opera phenix high content screening system, by PerkinElmer (5 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Cellcarrier, by PerkinElmer (2 mentions) E2c2515 ul scara robot, by Epson (2 mentions) D300 digital drug dispenser, by Hewlett-Packard (2 mentions) Stainless steel pins, by V&P Scientific (2 mentions) L2020, by Merck Group (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Synergy h1 plate reader, by Agilent Technologies (2 mentions) Prism 6, by GraphPad (2 mentions) Harmony software, by PerkinElmer (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Growth factor reduced matrigel, by Corning (1 mentions) A13777, by Thermo Fisher Scientific (1 mentions) Opera qehs spinning disk microscope, by PerkinElmer (1 mentions) Essential 8 media, by Thermo Fisher Scientific (1 mentions) El406 microplate washer dispenser, by Agilent Technologies (1 mentions) Janus automated workstation, by PerkinElmer (1 mentions)

36 protocols using cellcarrier ultra plates

1

Pancreatic Acinar Cell Organoid Isolation

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Acinar cells were isolated as previously reported (da Silva et al., 2022 (link)) from the pancreas of 6–8-week-old wild type (ADM inhibition assays), Cre-mice (ADM reversal assays), or KC mice (ADM reversal assays). Following a 30-min digestion with collagenase P, enzyme inactivation with fetal bovine serum containing Hanks’ balanced salt solution, and multiple centrifugation steps, cells were passed through a series of strainers (500, 300, and 200 µm). The resulting pellet was resuspended in media to the desired density to yield 50 organoids/well with growth factor reduced Matrigel (Corning). The mixture was maintained on ice during the pipetting steps. 20 µL of the mixture was pipetted into a 384-well CellCarrier Ultra plates (PerkinElmer), followed by 30 min of incubation to solidify the mixture and then 40 µL of warm media was added. The organoids seeded in Matrigel were left overnight to acclimate and then subjected to treatment or left to further develop ADM before treatment.
Atanasova K.R., Perkins C.M., Ratnayake R., Jiang J., Chen Q.Y., Schmittgen T.D, & Luesch H. (2024). Epigenetic small-molecule screen for inhibition and reversal of acinar ductal metaplasia in mouse pancreatic organoids. Frontiers in Pharmacology, 15, 1335246.
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2

Visualizing hiPSC gene-edited cellular dynamics

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Generation of the hiPSCs lines, imaging obtainment was performed as previously described13 (link). The human iPSCs line used in this study is A13777 obtained from Gibco. Briefly, the hiPSCs gene-edited with the Rosella construct into the AAVS1 safe harbour were cultured in Essential-8 media (Thermo Fisher cat no. A1517001) in CellCarrier Ultra plates (Perkin Elmer, 6055300). Confocal images were obtained with an Opera QEHS spinning disk microscope (Perkin Elmer) under a 60x water immersion objective ( NA·=·1.2 ). DsRed and pHluorin images were acquired simultaneously using two cameras and binning 2.
Garcia Santa Cruz B., Slter J., Gomez-Giro G., Saraiva C., Sabate-Soler S., Modamio J., Barmpa K., Schwamborn J.C., Hertel F., Jarazo J, & Husch A. (2022). Generalising from conventional pipelines using deep learning in high-throughput screening workflows. Scientific Reports, 12, 11465.
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3

High-throughput Compound Screening Assay

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Drugs were arrayed in nine-point half-log dilution series in 384-well library plates (Supplementary Data 5). The identity and purity of the drugs were verified by LC-MS. Each daughter plate contained 10 µl per well, and was thawed a maximum of 12 times. Cells were seeded in 384-well CellCarrier or CellCarrier ULTRA plates (catalog number 6057300, Perkin Elmer, Waltham, MA) with a multidrop combi liquid dispenser at the densities listed in Supplementary Data 4, and allowed to adhere for 24–36 h prior to drug treatment. Drugs were delivered from library plates via pin transfer with a custom E2C2515-UL Scara robot (Epson, Long Beach, CA) coupled to stainless steel pins (V&P Scientific, San Diego, CA) at the ICCB-Longwood Screening Facility, or from stock solutions with a D300 digital drug dispenser (Hewlett-Packard, Palo Alto, CA). At the time of drug delivery, replicate plates were stained and fixed to serve as time = 0 reference data for GR-based calculations, and 72 h later treated plates were stained and fixed according to the Dye Drop or Deep Dye Drop protocol (see below).
Mills C.E., Subramanian K., Hafner M., Niepel M., Gerosa L., Chung M., Victor C., Gaudio B., Yapp C., Nirmal A.J., Clark N, & Sorger P.K. (2022). Multiplexed and reproducible high content screening of live and fixed cells using Dye Drop. Nature Communications, 13, 6918.
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4

High-throughput Screening of OPC Differentiation Modulators

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Following expansion of OPCs (see OPC Cell Culture), OPCs were seeded onto 384-well CellCarrierUltra plates (PerkinElmer) coated with poly-D-lysine (15 μg/mL; 37°C for 1 h) and laminin (15 μg/mL; 37°C for 1 h) at a concentration of 10,000 cells per well in 50 μL of differentiation-permissive media. An EL406 Microplate Washer Dispenser (Biotek) equipped with a 5 μL dispensing cassette (Biotek) was used to dispense cells. A 100 μM stock of each drug in the bioactive library (Selleck Bioactive Compound Library-I; L1700) dissolved in dimethyl sulfoxide (DMSO) was prepared in Abgene storage 384-well plates (ThermoFisher Scientific). Drugs were added to the plates using a 50 nL solid pin tool attached to a Janus automated workstation (PerkinElmer), for a final screening concentration of 100 nM. DMSO was run as a negative control for each plate. A previously reported OPC differentiation-enhancing drug, TASIN-1, was used as a positive control at a concentration of 1 μM.15 (link) Cells were incubated at 37°C for 72 h and subsequently fixed and stained for MBP and DAPI according to the protocol in OPC Phenotypic Assay for MBP Positivity. Imaging and analysis of the plates was performed as outlined in High-Content Imaging and Analysis.
Sax J.L., Hershman S.N., Hubler Z., Allimuthu D., Elitt M.S., Bederman I, & Adams D.J. (2022). Enhancers of human and rodent oligodendrocyte formation predominantly induce cholesterol precursor accumulation. ACS chemical biology, 17(8), 2188-2200.
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5

Luciferase Reporter Assay for OPCs

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The luciferase reporters were acquired from Addgene: ABCA1 promoter plasmid (Plasmid #86442), TOPflash (Plasmid #12456), and FOPflash (Plasmid #12457). (Veeman et al., 2003 (link), Lai et al., 2018 (link)) 1 ug of DNA was delivered to OPCs using the Nucleofector Kit for Primary Mammalian Glial Cells and the Nucleofector 2b Device by following standard manufacturer’s protocol. Transformed OPCs were seeded at 50,000 cells per well on 96-well CellCarrierUltra plates (PerkinElmer) coated with poly-D-lysine and laminin (Sigma, L2020; 15 μg/mL) using multi-channel pipet and treated with small molecules. Cells were incubated under standard conditions (37° C, 5% CO2) for 2 days. Cells were lysed using the Bright-Glo luciferase assay system (Promega, E2620) and were analyzed using a Synergy Neo2 High Performance plate reader.
Hubler Z., Friedrich R.M., Sax J., Allimuthu D., Gao F., Rivera-León A.M., Pleshinger M., Bederman I, & Adams D.J. (2021). Modulation of lanosterol synthase drives 24,25-epoxysterol synthesis and oligodendrocyte formation.. Cell chemical biology, 28(6), 866-875.e5.
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6

Long-term Neuronal and Astrocyte Culture

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Neurons and astrocytes were cultured in 384- or 96-well Cell Carrier Ultra plates (PerkinElmer) in Neuron Maintenance Medium and half of the volume of culture media was changed using Tecan Fluent automation liquid-handling workstation every 3–4 days for at least 8 weeks and up to 6 months before subsequent experimentation. Tecan Fluent was programmed to utilize its features of automated tip loading, lid removal, and to aspirate half the volume of culture media and add new culture media for up to 30 plates at a time. Barcode-operated plate storage incubator technology was integrated into the system for plate organization and retrieval.
Bassil R., Shields K., Granger K., Zein I., Ng S, & Chih B. (2021). Improved modeling of human AD with an automated culturing platform for iPSC neurons, astrocytes and microglia. Nature Communications, 12, 5220.
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7

Organoid Imaging for Chromocenters

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Organoids were cultured in black 96-well CellCarrier Ultra plates suitable for high content imaging (Perkin Elmer, Hamburg, Germany, cat. no. 6055302) and incubated with Hoechst 33342 (2 µg/mL, Invitrogen, Thermo Fisher Scientific, cat. no. H3570) for 3 h prior to imaging with the Opera Phenix High Content Screening System (Perkin Elmer) equipped with a 40× water immersion objective and a 16-bit sCMOS 4 Megapixel camera. Hoechst was excited with a 405 nm solid-state laser line and detected at 435–480 nm wavelength range. Organoids were imaged in the hydrogel dome, using 100 plane Z-stacks with a step size of 2 µm between planes. Nuclei in organoids of at least 50 µm in diameter were manually assessed for the presence of chromocenters, that are visible as high-intensity nuclear foci.
Van Hemelryk A., Tomljanovic I., de Ridder C.M., Stuurman D.C., Teubel W.J., Erkens-Schulze S., Verhoef E.I., Remmers S., Mahes A.J., van Leenders G.J., van Royen M.E., van de Werken H.J., Grudniewska M., Jenster G.W, & van Weerden W.M. (2022). Patient-Derived Xenografts and Organoids Recapitulate Castration-Resistant Prostate Cancer with Sustained Androgen Receptor Signaling. Cells, 11(22), 3632.
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8

Transfection and Immunodetection of CGRP Receptor

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HEK293S cells were cultured and transfected as previously described (Gingell et al., 2020 (link)). Cells were plated into poly-D-lysine coated Cell-Carrier Ultra plates (PerkinElmer, Waltham, MA) at 10,000 cells per well and transfected 36 h after plating with 0.25 µg of DNA, using polyethylenimine as previously described (Gingell et al., 2020 (link)). Immunocytochemistry was performed 24–36 h after transfection. For western blotting, HEK293S cells were grown in 15 cm2 dishes and transfected with 60 µg of DNA when ∼70% confluent. Cells were harvested for whole cell lysate preparations 48 h after transfection. In all cases, CLR or CTR were transfected in a 1:1 ratio with either RAMP1 or pcDNA3.1. Rat, mouse and human CT(a) splice variants were used as controls for immunocytochemistry and immunoblotting. However, the antigenic sequences for mAb8B9, pAb188 and mAb31-01 are present in both the CT(a) and CT(b) variants. Therefore, these antibodies are expected to detect both variants.
Rees T.A., Russo A.F., O’Carroll S.J., Hay D.L, & Walker C.S. (2022). CGRP and the Calcitonin Receptor are Co-Expressed in Mouse, Rat and Human Trigeminal Ganglia Neurons. Frontiers in Physiology, 13, 860037.
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9

Fluorescent Virus Infection Assay

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All viruses expressing fluorescent (GFP, ZsGreen) proteins were assayed for fluorescence by using an H1 Synergy plate reader (Biotek). NCI-H358 cells or SAECs were seeded at 2 × 104 cells per well in black opaque 96-well plates (Corning 3619, Corning, NY) or Perkin-Elmer CellCarrier Ultra plates (Waltham, MA) and compounds were added to the assay plates for 1 h. Assay plates were transferred to the BSL-4 suite (where appropriate), and infected with 0.25–0.5 TCID50 per cell of the respective virus, and were read between 48 and 168 h post-infection (hpi) depending on the virus used. Fluorescence signal from DMSO-treated infected cells were set as 100% GFP. 50% effective concentrations (EC50) were calculated using four-parameter variable slope non-linear regression fitting of mean values of assays performed in quadruplicate (Graphpad Prism 6, La Jolla, CA).
Lo M.K., Jordan P.C., Stevens S., Tam Y., Deval J., Nichol S.T, & Spiropoulou C.F. (2018). Susceptibility of paramyxoviruses and filoviruses to inhibition by 2′-monofluoro- and 2′-difluoro-4′-azidocytidine analogs. Antiviral Research, 153, 101-113.
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10

Monitoring Autophagy in Macrophages

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RAW264.7 macrophages were electroporated with Neon (Life Technologies, Thermo Fisher Scientific) to express GFP-mCherry-LC3B (a gift from Sharon Tooze, The Francis Crick Institute, London, United Kingdom). Cells were washed in PBS and resuspended at 1.5 × 106 cells in 100 μL Buffer R with 5 μg plasmid DNA before electroporating in Buffer E at 1,680 V for 20 ms and plated at 1 × 105 cells per well in 96-well CellCarrier Ultra plates (PerkinElmer) overnight. Cells were washed twice in PBS and then stimulated with DMSO, 100 nM BafA1, and 10 μM TbAd for 4 hours before fixing in 4% PFA. Nuclei were counterstained with DAPI and more than 900 transfectants per condition were imaged (Opera Phenix).
Bedard M., van der Niet S., Bernard E.M., Babunovic G., Cheng T.Y., Aylan B., Grootemaat A.E., Raman S., Botella L., Ishikawa E., O’Sullivan M.P., O’Leary S., Mayfield J.A., Buter J., Minnaard A.J., Fortune S.M., Murphy L.O., Ory D.S., Keane J., Yamasaki S., Gutierrez M.G., van der Wel N, & Moody D.B. (2023). A terpene nucleoside from M. tuberculosis induces lysosomal lipid storage in foamy macrophages. The Journal of Clinical Investigation, 133(6), e161944.
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