Cytation 1

Manufactured by Agilent Technologies

Sourced in United States

The Cytation 1 is a compact and versatile cell imaging multi-mode reader designed for a wide range of cell-based assays. It combines automated digital microscopy with multi-mode detection capabilities to provide quantitative data for your research.

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Lab products found in correlation

Gen5 microplate data collection and analysis software, by Agilent Technologies (16 mentions) Hoechst 33342, by Thermo Fisher Scientific (6 mentions) Ros assay kit, by Beyotime (3 mentions) Hk210 01, by Hycult Biotech (2 mentions) Infinite m200 pro, by Tecan (2 mentions) 96 well plate, by Corning (2 mentions) Axio observer d1, by Zeiss (2 mentions) Hoechst 33342 staining assay, by Thermo Fisher Scientific (2 mentions) Xfe96 analyzer, by Agilent Technologies (2 mentions) Co2 gas controller, by Agilent Technologies (1 mentions) 24 well ultra low attachment plate, by Corning (1 mentions) Atp determination kit, by Thermo Fisher Scientific (1 mentions) 6 well plate, by Corning (1 mentions) Caspase glo 3 7 reagent, by Promega (1 mentions) Celltox green dye, by Promega (1 mentions) Ldh storage buffer, by Promega (1 mentions) Gen5 data analysis software, by Agilent Technologies (1 mentions) Prism 8, by GraphPad (1 mentions) Renilla luciferase assay system, by Promega (1 mentions) Edu incorporation assay kit, by RiboBio (1 mentions)

Close spelling variants of this product

Cytation 1 Cell Imaging Multi-Mode Reader (65 citations) Cytation 1 plate reader (10 citations) Cytation 1 imaging reader (10 citations) BioTek Cytation 1 (8 citations) Cytation 1 imaging multimode reader (6 citations) Cytation 1 multi-mode imaging system (5 citations) Cytation 1 cell imaging reader (5 citations) Cytation 1 Cell Imaging System (5 citations) BioTek Cytation 1 cell imaging multi-mode reader (3 citations) Cytation 1 Imager (3 citations)

74 protocols using cytation 1

Live-cell Imaging of Mitochondrial pH

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For live-cell time-lapse imaging, cells are switched to phenol red free Imaging media as described (Section 2.1). Time-lapse images were taken every two hours on a Cytation 1 (BioTek). The Cytation 1 was set to 37 °C, and a CO₂ gas controller (BioTek) was set to 5%. Cells were imaged at 20x magnification, allowing for the use of clear bottomed 96-well plates (Costar), and the ability to image ~ 20 cells per field, with multiple images taken per well over time. These conditions increased our ability to conduct high-throughput analysis on a large number of donors, and more accurately quantify each donor’s mKeima-mito signal by increasing the number of cells evaluated.
A custom filter cube was obtained from BioTek to detect mKeima’s unique neutral pH excitation wavelength (Table 1). Under a neutral pH, mKeima excites at 440nm and emits at 610nm. For neutral detection, a 465nm LED, 445/45nm excitation filter, and 647/57nm emission filter (BioTek) is used. Under an acidic pH, mKeima excites at 580nM and emits at 610nM. For detecting acidic pH mitochondria a standard Texas Red filter set (BioTek) was used. This cube has a 590nm LED, 586/15nm excitation filter, and 647/57nm emission filter.

Fisher C.R., Ebeling M.C, & Ferrington D.A. (2022). Quantification of mitophagy using mKeima-mito in cultured human primary retinal pigment epithelial cells. Experimental eye research, 217, 108981.

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Morphological Analysis of H9c2(2-1) and HepaRG Cells

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To verify the influence of LB-FA and LB-FT associations on the morphology and confluence of H9c2(2-1) and HepaRG cells, a bright field microscopic examination was performed. The cells were photographed at the end of the 72-h treatment period using Cytation 1 (BioTek Instruments Inc., Winooski, VT, USA). The obtained pictures were processed using the Gen5™ Microplate Data Collection and Analysis Software (BioTek Instruments Inc., Winooski, VT, USA).

Rednic R., Marcovici I., Dragoi R., Pinzaru I., Dehelean C.A., Tomescu M., Arnautu D.A., Craina M., Gluhovschi A., Valcovici M, & Manea A. (2022). In Vitro Toxicological Profile of Labetalol-Folic Acid/Folate Co-Administration in H9c2(2-1) and HepaRG Cells. Medicina, 58(6), 784.

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Quercetin Effects on Cell Proliferation

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Cells were seeded into 96-well plates at a density of 8 × 10³ cells/well and grown in McCoy’s 5A medium (plus 10% FBS). Cells in each well were treated with quercetin at different concentrations (0, 10, 20, 40, 80, and 160 μM, n = 5) for 24 or 48 h after the cell confluence reached about 100%. Then, these cells were incubated with CCK-8 solution to determine the cell proliferation rate (%). The optical density (OD) was measured at 450 nm in a microplate reader (Cytation 1, BioTek) for further IC50 analysis.

Li X., Zhu Q., Ma M, & Guo H. (2022). Quercetin inhibits the progression of endometrial HEC-1-A cells by regulating ferroptosis—a preliminary study. European Journal of Medical Research, 27, 292.

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Electrochemotherapy ATP Determination

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For ATP determination, electrochemotherapy was performed as for the clonogenic assay. For the timepoints 0 and 4 h, the cells were incubated in 24 well ultra-low attachment plates (Corning Costar) and at the designated times 250 µL of cell culture medium was collected. For the 24 and 48 h timepoints, the cells were seeded on 6 well plates (Corning Costar) after electrochemotherapy and at the designated timepoints 250 µL of cell culture medium was collected. The samples were then centrifuged (300× g, 4 °C, 5 min) and 10 µL of the supernatant was used in the ATP Determination Kit (Invitrogen, Thermo Fisher Scientific) assay following the manufacturer’s instructions. The resulting luminescence was measured with Cytation 1 (BioTek), and the final ATP concentration was calculated from the obtained standard curve.

Kesar U., Markelc B., Jesenko T., Ursic Valentinuzzi K., Cemazar M., Strojan P, & Sersa G. (2023). Effects of Electrochemotherapy on Immunologically Important Modifications in Tumor Cells. Vaccines, 11(5), 925.

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ELISA Antibody Titer Determination

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ELISA was performed according to a previously report protocol with minor modifications [47 (link)]. ELISA plates were incubated with conjugate 2a, 2b or 2c (2 μg/mL, 100 μL per well) in coating buffer (0.1 M bicarbonate, pH 9.6) at 4 °C overnight and then at 37 °C for 1 h. After washing with PBST (PBS containing 0.05% of Tween-20) 3 times, the plates were treated with blocking buffer (1% BSA in PBST) at rt for 1 h. Pooled antisera diluted from 1:300 to 1:72900 in PBS in serial half-log manner were added to the plates (100 μL/well), which was followed by incubation at 37 °C for 2 h. Then, AP-linked goat anti-mouse kappa, IgG and IgM antibodies (1:1,000 dilution in PBS) were added and the plates were incubated at rt for 1 h. After washing with PBST 3 times, 100 μL of a p-nitrophenylphosphate (PNPP) solution (1.67 mg/mL in PBS) were added and the plates were then incubated at rt for 30 min. The optical density (OD) values of the ELISA plates at 405 nm wavelength were read using a microplate reader (Cytation 1, Bio-Tek instruments Inc.). OD values were plotted against the serum dilution numbers to obtain a best-fit logarithm line and the equation of this line was used to calculate the antibody titer. The dilution number at which the OD value of 0.2 was achieved was taken as the antibody titer.

Guo J., Jiang W., Li Q., Jaiswal M, & Guo Z. (2020). Comparative Immunological Studies of Tumor-Associated Lewis X, Lewis Y, and KH-1 Antigens. Carbohydrate research, 492, 107999.

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Hoechst Assay for Volatile Oil-Induced Cell Death

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To determine the type of cell death induced by volatile oils, the Hoechst method was performed. The applied protocol followed the manufacturer’s instructions. Briefly, cells were cultured at 1 × 10⁵/well in 12-well plates. After reaching a confluence of approximately 90%, the cells were treated with three different concentrations of EO: 5, 25, and 75 μg/mL. After 24 h of treatment, the medium was removed and 100 μL of staining solution was added to each well diluted 1:2000 in PBS. After incubation for 10 min at room temperature and protected from light, the staining solution was removed and washed three times with PBS. The pictures were taken using Cytation 1 (BioTek Instruments Inc., Winooski, VT, USA) and processed using the Gen5 Microplate Data Collection and Analysis Software (BioTek Instruments Inc., Winooski, VT, USA). The apoptotic index was calculated according to a formula described by Xu et al. [46 (link)] and used in one of our previous publications [47 (link)].

Dolghi A., Buzatu R., Dobrescu A., Olaru F., Popescu G.A., Marcovici I., Pinzaru I., Navolan D., Cretu O.M., Popescu I, & Dehelean C.A. (2021). Phytochemical Analysis and In Vitro Cytotoxic Activity against Colorectal Adenocarcinoma Cells of Hippophae rhamnodies L., Cymbopogon citratus (D.C.) Stapf, and Ocimum basilicum L. Essential Oils. Plants, 10(12), 2752.

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Ebola Virus Infection Assay

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Hela cells were preincubated with each small molecule compound for 1 hour before being challenged with wild type Ebola virus Zaire at an MOI of 0.1 to 0.3. After 24 hours, cells were fixed in formalin and removed from the BSL4 lab for staining with virus specific monoclonal antibody against GP (4F3, IBT Bioservices, MD, USA). After staining for virus antigen with primary and Alexa488-labeled anti-mouse secondary antibody and cell nuclei using Hoechst 33342) wells were imaged using an automated imaging microscope (Cytation 1, Biotek, VT, USA). Images were processed using CellProfiler (Broad Inst, MA, USA) to count infected and total cells using a custom pipeline that is available upon request to RAD. Infection efficiency was calculated as the ratio of infected cells to total cells. Experiments were performed with at least 3 replicates.

Wang L.L., Palermo N., Estrada L., Thompson C., Patten J., Anantpadma M., Davey R.A, & Xiang S.H. (2021). Identification of Filovirus Entry Inhibitors Targeting the Endosomal Receptor NPC1 Binding Site. Antiviral research, 189, 105059.

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Efflux Inhibitors Enhance Antimicrobial Efficacy

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Strains 1199B and K2068 were seeded on Heart Infusion Agar (HIA) 24 h before the experiments and kept in a bacteriological oven at 37 °C. The inoculum was prepared in PBS until obtaining 1.5 × 10⁸ colony forming units (CFU), corresponding to the 0.5 value on the McFarland scale. The 1199B inoculum was prepared in a test tube by adding valencene or nootkatone at 100 µg/mL. The K2068 strain inoculum was added with valencene or nootkatone at 50 µg/mL in another group. The CCCP group (50 µg/mL) was used as a positive control for both strains. The negative control consisted only of the inoculum. In all tubes, PBS was added until reaching a final volume of 1 mL. After 90 min of incubation, EtBr (100 µg/mL) was added to all groups except the growth control. The solutions were again incubated for 1 h, then centrifuged at 10,000 rpm for 2 min, and washed twice with interspersed centrifugation cycles to remove EtBr and any supernatant substance. After centrifugation, the pellet was homogenized in PBS and distributed in the wells on a microplate. The reading was performed at 10, 20, 30, 40, 50, 60, and 70 min, using a fluorescence microplate reader (Cytation 1, BioTek^®, Winooski, VT, USA) and Gen5™ 3.22 Software, with excitation of 530 nm and wavelength of 590 nm. The assay was performed in triplicate [24 (link),25 (link),26 (link)].

Oliveira-Tintino C.D., Santana J.E., Alencar G.G., Siqueira G.M., Gonçalves S.A., Tintino S.R., de Menezes I.R., Rodrigues J.P., Gonçalves V.B., Nicolete R., Ribeiro-Filho J., da Silva T.G, & Coutinho H.D. (2023). Valencene, Nootkatone and Their Liposomal Nanoformulations as Potential Inhibitors of NorA, Tet(K), MsrA, and MepA Efflux Pumps in Staphylococcus aureus Strains. Pharmaceutics, 15(10), 2400.

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Multiplex Cytotoxicity Assay for Compounds

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As contrast to activity, toxicity of top six similars and lead compounds was determined by a multiplex measurement of caspase activation and membrane integrity (using LDH release and inclusion of the CellTox Green dye). Briefly, HCFs were seeded as before. After 24 h, cells were treated with serial dilutions of compounds with additional 0.1% CellTox™ Green dye (Promega, #G8741). After 24 h incubation, 5 µL medium was removed from each well, mixed with 95 µL LDH Storage buffer (Promega, #J2380) and frozen for later analysis. 95 µL Caspase-Glo® 3/7 reagent (Promega, #G8090) was added to each well, and after 60 min of incubation luminescence was measured in a Synergy HT. Afterwards, micrographs of brightfield and green fluorescence were taken using a Cytation 1 (BioTek) automated microscope maintaining 37 °C and 5% CO₂. Wells were manually checked for co-localization of fluorescence signal with cells in brightfield. On the next day, supernatants in LDH Storage buffer were thawed, and 50 µL was mixed with 50 µL of LDH Detection reagent and luminescence was measured after 60 min of incubation. Dose–response curves were determined as described before. To determine toxicity of the top 3 anti-fibrotic similars in hiPS-CM, 20,000 cells/well were seeded, treated and analysed as above, devoid the fluorescent dye steps.

Kreutzer F.P., Meinecke A., Mitzka S., Hunkler H.J., Hobuß L., Abbas N., Geffers R., Weusthoff J., Xiao K., Jonigk D.D., Fiedler J, & Thum T. (2022). Development and characterization of anti-fibrotic natural compound similars with improved effectivity. Basic Research in Cardiology, 117(1), 9.

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Kinetic Evaluation of Electroporation-Induced Cell Death

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Due to the minimal or no effect of bleomycin or cisplatin alone on cell viability, cell death kinetics were determined only in cells treated with EP and ECT. Cell death was determined through kinetic evaluation of diamidino-2-phenylindole (DAPI, Thermo Fisher Scientific) uptake, determining increased cell membrane permeability as a marker of cell death. Briefly, ECT with 0.14 μM bleomycin was evaluated alone or combined with 2.5, 5 and 7.5 μM sunitinib. Afterwards, 5 x 10³ control or EP-treated cells and 25 x 10³ ECT-treated cells were seeded in each well of a black 96-well plate. Different cell plating densities were used because of prominent cell death that follows instantly after ECT. These dead cells do not attach to the plate and represent a limitation for the usage of imaging techniques. Cells were incubated for 2 hours to allow cells to attach. Thereafter, medium with non-attached dead cells was removed for cell imaging purposes, and 200 μL of fresh phenol-free medium containing the appropriate concentration of sunitinib and 0.5 μM DAPI was added. Brightfield and fluorescence images (one image per well) of cells were captured every 4 h for 48 hours with Cytation 1 (BioTek) with a 4-x objective. For each time point, the number of DAPI-positive nuclei (representing dead cells) was determined with Gen 5 Data analysis software (BioTek).

Bosnjak M., Jesenko T., Markelc B., Cerovsek A., Sersa G, & Cemazar M. (2022). Sunitinib Potentiates the Cytotoxic Effect of Electrochemotherapy in Pancreatic Carcinoma Cells. Radiology and Oncology, 56(2), 164-172.

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