Cellsens dimension 1

Manufactured by Olympus

Sourced in Japan, Germany

CellSens Dimension 1.11 software is a digital imaging and analysis solution for microscopy. It provides core functionality for image acquisition, processing, and analysis. The software is designed to work with a range of Olympus microscope systems.

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119 protocols using cellsens dimension 1

Quantifying B-Cell Infiltration in Organs

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All paraffin-embedded organs were stained with H&E to perform a complete histopathological analysis and a clinical pathologist supervised all samples for toxicity evaluation. To detect the presence of human B-cells and to quantitate the percentage of organ infiltration, we used immunohistochemistry (IHC) staining with an antibody anti-human CD20cy (clone L26, Dako). Firstly, 5 and 27 low power fields at 100 magnifications were taken for LNs and BM, respectively, in each mice group (buffer or T22-PE24- H6). Secondly, the area occupied by B cells in each organ was selected and the percentage of CD20+ cells was quantified using the cellSens Dimension 1.9 software (Olympus).
IHC staining was performed in a DAKO Autostainer Link48 (Agilent) following the manufacturer's instructions. Representative pictures were taken using an Olympus DP73 digital camera and processed with the cellSens Dimension 1.9 software (Olympus) at 200 or 400 magnifications.

Falgàs A., Pallarès V., Serna N., Sánchez-García L., Sierra J., Gallardo A., Alba-Castellón L., Álamo P., Unzueta U., Villaverde A., Vázquez E., Mangues R, & Casanova I. (2020). Selective delivery of T22-PE24-H6 to CXCR4+ diffuse large B-cell lymphoma cells leads to wide therapeutic index in a disseminated mouse model. Theranostics, 10(12), 5169-5180.

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Wound Closure Assay with Adhesive Inserts

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Adhesive wound assay inserts (Ibidi) were used to generate wounds in confluent layers in a 6 well plate. To do this, transfected cells were trypsinized and seeded into the plate to form a monolayer. After overnight growth and attachment, the insert was carefully removed and the cells were starved of serum by adding serum free media for 4 hours, after which DMEM was added to the cells and they were imaged immediately (T0) using (Olympus cellSens Dimension 1.12). Cells were maintained in an incubator at 37°C, 5% CO₂ for 24 hours, after which time images of the wound closure were taken as described above. The percentage wound closure was analysed using Wimasis image analysis.

Dowling C.M., Hayes S.L., Phelan J.J., Cathcart M.C., Finn S.P., Mehigan B., McCormick P., Coffey J.C., O'sullivan J, & Kiely P.A. (2017). Expression of protein kinase C gamma promotes cell migration in colon cancer. Oncotarget, 8(42), 72096-72107.

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3D Cell Culture Assay for Angiogenesis

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Individual wells of a 6 well plate were coated with Matrigel^™ (BD Biosciences) and placed in an incubator at 37^°C for 30 min. Stable cell lines were trypsinized and counted. 50,000 cells/ml were resuspended in DMEM supplemented with 2% Matrigel^™. Cells were placed in Matrigel^™ coated wells for 30 min at 37^°C, after which DMEM supplemented with 2% Matrigel^™ was added to the cultures. Cells were maintained in culture for 6 days in an incubator at 37^°C, 5% CO₂ with fresh medium added every 2 days and cultures imaged every 24 hours (Olympus cellSens Dimension 1.12). On day 6, cultures were harvested using EDTA/PBS, fixed with paraformaldehyde (PFA) and stained for confocal analysis (Zeiss LSM 710).

Dowling C.M., Phelan J., Callender J.A., Cathcart M.C., Mehigan B., McCormick P., Dalton T., Coffey J.C., Newton A.C., O'sullivan J, & Kiely P.A. (2016). Protein kinase C beta II suppresses colorectal cancer by regulating IGF-1 mediated cell survival. Oncotarget, 7(15), 20919-20933.

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Optimized Liquid Overlay for 3D MCTS

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The present study typically optimized the liquid overlay technique to acquire single and stable 3D MCTSs with spherical structure and uniform size. The 96-well plates were coated with agar gel, which prevents cell attachment and results in the cells suspension aggregating into cell spheroids. Briefly, 1% agar gel (Sigma-Aldrich; Merck KGaA) was processed by high-pressure-steam sterilizing and melted in microwaves prior to use. The hot agar solution was immediately pipetted into a 96-well plate (65 µl/well) and cooled for ~30 min to solidify. Next, T24 and 5637 single cell suspensions were added to each well with different initial seeding densities (500–10,000/well) and cultured in an incubator for 24 h. The culture medium was changed every 2 days the growth characteristics of spheroids were observed and recorded. Only one MCTS formed per well at 24 h with a spherical structure and uniform size was selected for further experiments. The field microscopy images of MCTSs at different initial seeding densities were acquired using an inverted microscope (IX73; Olympus Corporation, Tokyo, Japan). The diameters of each spheroid were measured simultaneously using CellSens Life Science Imaging Software (CellSens Dimension 1.12; Olympus Corporation, Tokyo, Japan).

Zhuo Z., Song Z., Ma Z., Zhang Y., Xu G, & Chen G. (2019). Chlorophyllin e6-mediated photodynamic therapy inhibits proliferation and induces apoptosis in human bladder cancer cells. Oncology Reports, 41(4), 2181-2193.

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3D Cell Culture in Matrigel Assay

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Individual wells of a 6 well plate were coated with Matrigel^TM (BD Biosciences) and placed in an incubator at 37°C for 30 min. Transfected cells were trypsinized and counted. 50,000 cells/ml were resuspended in DMEM supplemented with 2% Matrigel^TM. Cells were placed in Matrigel^TM coated wells for 30 min at 37°C, after which DMEM supplemented with 2% Matrigel^TM was added to the cultures. Cells were maintained in culture for 6 days in an incubator at 37°C, 5% CO₂ with fresh medium added every 2 days and cultures imaged every 24 hours (Olympus cellSens Dimension 1.12).

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Wound Healing and Directional Migration Assays

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For wound healing assays, transfected cells were seeded into cell culture wells containing ibidi inserts (ibidi GmbH, Gräfelfing, Germany). After 72 h incubation, the insert was carefully removed and cells imaged using Olympus cellSens Dimension 1.12. Cells were incubated for 24 h, after which images of the wound closure were taken. Six fields of view were imaged for each condition and percent wound closure was measured using Image J (version 1.52n). Three individual experiments were carried out to verify results. Data is displayed as mean across three individual experiments +/− SEM.
For directional migration assays, the underside of 8 µm transwell membranes were coated with fibronectin before 6 × 10⁵ cells in serum free DMEM media were plated onto the upper chamber. Cells were allowed to migrate towards a lower chamber containing 50 ng/mL EGF in serum free media for 24 h. Cells were removed from the upper chamber of the membrane. Migrated cells were fixed and stained with crystal violet and imaged. To quantify the cell migration, crystal violet stain was dissolved 10% acetic acid and intensity measured as absorbance at 595 nm in triplicate. Three individual experiments were carried out to verify results.

Mahdi A.F., Malacrida B., Nolan J., McCumiskey M.E., Merrigan A.B., Lal A., Tormey S., Lowery A.J., McGourty K, & Kiely P.A. (2020). Expression of Annexin A2 Promotes Cancer Progression in Estrogen Receptor Negative Breast Cancers. Cells, 9(7), 1582.

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Ultrastructural Analysis of Hippocampal DG

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An Olympus IX83 inverted microscope (Olympus, Hamburg, Germany) coupled to an image capture system (CellSens Dimension 1.12., Olympus, Hamburg, Germany) was used for a qualitative evaluation of the semithin sections. Ultrathin sections (70 nm) were used to evaluate the DG hippocampal subregion and were examined using a JEM-1011 electronic microscope. Selected images were collected with an ES1000W Erlangshen CCD camera (GATAN Inc., Pleasanton, CA, USA) at uniform magnification, resulting in 2194 µm² fields. Data collection and measurements were performed in a blinded manner. The integrity of the cell structures and the presence or absence of degenerate cells were analyzed. In addition, 15 cells without the rupture of both nuclear and cytoplasmic membranes were randomly selected from each animal (n = 2/genotype). Ultrastructural alterations were classified according to the following semi-quantitative classification system: − (absent), + (mild), ++ (moderate), and +++ (severe).

de Paula Nascimento-Castro C., Winkelmann-Duarte E.C., Mancini G., Welter P.G., Plácido E., Farina M., Gil-Mohapel J., Rodrigues A.L., de Bem A.F, & Brocardo P.S. (2022). Temporal Characterization of Behavioral and Hippocampal Dysfunction in the YAC128 Mouse Model of Huntington’s Disease. Biomedicines, 10(6), 1433.

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Wound Closure Kinetics Assay

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Adhesive wound assay inserts (Ibidi) were used to generate wounds in confluent layers in a 6 well plate. To do this, stable cells were trypsinized and seeded into the plate to form a monolayer. After overnight growth and attachment, the insert was carefully removed and the cells were starved of serum by adding serum free media for 4 hours, after which DMEM was added to the cells and they were imaged immediately (T0) using (Olympus cellSens Dimension 1.12). Cells were maintained in an incubator at 37^°C, 5% CO₂ for 24 hours, after which time images of the wound closure were taken as described above. The percentage wound closure was analyzed using Wimasis image analysis.

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Myogenic Differentiation Imaging Workflow

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Images of myogenic differentiation were taken using Olympus cellSens Dimension 1.12. Copyright© 2009–2014 OLYMPUS CORPORATION Core Version XV 3.11 (Build 12849) Package number 5900 (cellSens Dimension). Labelling and detection probes Nucblue live stain (R37605) and Actin green 488 (R37110) was purchased from Life Technologies. Images were saved as JPEG images and analysed using ImageJ to measure changes in myotube thickness.

Murphy S.M., Kiely M., Jakeman P.M., Kiely P.A, & Carson B.P. (2016). Optimization of an in vitro bioassay to monitor growth and formation of myotubes in real time. Bioscience Reports, 36(3), e00330.

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3D Cell Culture and Drug Screening

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Individual wells of a 12 well plate were coated with Matrigel (Corning) and placed in an incubator at 37°C for 30 min. A total of 10,000 cells/ml were resuspended in RPMI supplemented with 2% Matrigel. Cells were placed in Matrigel -coated well for 15 min at 37°C, after which RPMI supplemented with 2% Matrigel was added to the cultures. Cells were treated with indicated drugs after 24 hours. Cells were maintained in culture for 7 days in an incubator at 37°C, 5% CO 2 , and cultures were imaged every 48 hours with a Olympus cellSens Dimension 1.12.

Zhang H., Nabel C.S., Li D., O'Connor R.Í., Crosby C.R., Chang S.M., Hao Y., Stanley R., Sahu S., Levin D.S., Chen T., Tang S., Huang H.Y., Meynardie M., Stephens J., Sherman F., Chafitz A., Costelloe N., Rodrigues D.A., Fogarty H., Kiernan M.G., Cronin F., Papadopoulos E., Ploszaj M., Weerasekara V., Deng J., Kiely P., Bardeesy N., Vander Heiden M.G., Chonghaile T.N., Dowling C.M, & Wong K.K. (2023). Histone Deacetylase 6 Inhibition Exploits Selective Metabolic Vulnerabilities in LKB1 Mutant, KRAS Driven NSCLC. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 18(7).

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