Fcs express software package

Manufactured by De Novo Software

Sourced in United States

FCS Express is a software package designed for the analysis and visualization of flow cytometry data. The software provides tools for tasks such as data import, gating, and statistical analysis of flow cytometry samples.

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Lab products found in correlation

Facscalibur, by BD (6 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions) Edu cell proliferation assay kit, by Thermo Fisher Scientific (2 mentions) Isotype matched iggs, by Beckman Coulter (2 mentions) Chrompure human igg whole molecule, by Jackson ImmunoResearch (2 mentions) Cd105, by BioLegend (2 mentions) Annexin 5 fitc apoptosis detection kit, by Abcam (1 mentions) Cellvue claret, by Merck Group (1 mentions) Modfitlt version 3, by Verity Software House (1 mentions) H2dcfda, by Thermo Fisher Scientific (1 mentions) Cellquest pro software, by BD (1 mentions) Fluostar optima, by BMG Labtech (1 mentions) Facsverse, by BD (1 mentions) Annexin 5 fluorescein isothiocyanate fitc, by Merck Group (1 mentions) Dihydroethidium dhe, by Merck Group (1 mentions) Propidium iodide, by Merck Group (1 mentions) Cyflow cube 8, by Sysmex (1 mentions) Stage specific embryonic antigen 4, by BioLegend (1 mentions) Ssea4, by BioLegend (1 mentions) Integrin β5, by Cell Signaling Technology (1 mentions)

8 protocols using fcs express software package

Cell Proliferation and Apoptosis Assay

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Before cell expansion, PL3 cells were labeled with CellVue^® Claret (Sigma–Aldrich) at 2 × 10⁻⁶ M for 5 min according to the manufacturer's protocol. After 8 days, expanded cells were collected and measured using a BD FACS Calibur™ flow cytometer (dual laser) (BD Biosciences, San Jose, CA). Twenty thousand events of each sample were collected using CellQuest Pro software (BD Biosciences) and cell proliferation index was analyzed by ModFit LT™ version 3.1 (Verity Software House, Topsham, ME).
Apoptosis of expanded cells was detected using Annexin V-FITC Apoptosis Detection Kit (BioVision Inc., Milpitas, CA). Briefly, 2 × 10⁵ detached cells from each group (n = 3) were labeled with FITC conjugated annexin V and propidium iodide for 15 min. Samples were measured using FACS Calibur (BD Biosciences) and analyzed using the FCS Express software package (De Novo Software, Los Angeles, CA).
Expanded cells were incubated with 1 mM hydrogen peroxide (H₂O₂) at 37 °C for 1 h. To measure intracellular reactive oxygen species (ROS), cells were incubated with 1 μM 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCFDA) (Life Technologies) for 15 min. The plates were read on a FlUOstar OPTIMA (BMG Labtech Inc., Cary, NC) with an excitation wavelength of 485 nm and emission of 530 nm. Samples were assayed in triplicate.

Li J., He F, & Pei M. (2015). Chondrogenic priming of human fetal synovium-derived stem cells in an adult stem cell matrix microenvironment. Genes & Diseases, 2(4), 337-346.

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Quantifying Platelet GPIbα and GPVI Expression

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V450 Mouse Anti-Human CD41a antibody (CD41a-V450), BV510 Mouse Anti-Human CD42b antibody (CD42b-BV510) (both from BD Bioscience, San Jose, CA) and efluor 660-labeled anti-human GPVI (GPVI-efluor 660) (eBioscience, San Diego, CA) were used to identify platelets, and to quantify the receptor GPIbα and GPVI surface expression, respectively. The isotype controls for CD42b-BV510 and GPVI-efluor 660 were IgG1K-BV510 and IgG1K- efluor 660, respectively. To assess GPIbα and GPVI surface expression, 5 µl collected blood samples (baseline and sheared) were immediately incubated with 5 µl CD42b-BV510 or IgG1K-BV510, 5 µl GPVI-efluor 660 or IgG1K-eFluor 660, 5 µl CD41a-V450 and 25 µl HEPES buffer. After 30 min incubation at RT in the dark, the samples were fixed with 1 ml of 1% paraformaldehyde (PFA) in phosphate buffered saline (PBS) for 30 min at 4^oC in the dark. The flow cytometry data acquisition was performed on a flow cytometer (FACSVerse, BD Bioscience, San Jose, CA). FCS express software package (De Novo Software, Glendale, CA) was used to analyze flow cytometry data.

Chen Z., Tran D., Li T., Arias K., Griffith B.P, & Wu Z.J. (2020). The role of ADAM proteolysis and mechanical damage in non-physiological shear stress induced platelet receptor shedding. ASAIO journal (American Society for Artificial Internal Organs : 1992), 66(5), 524-531.

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Evaluating Cell Proliferation in Normoxia and Hypoxia

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Levels of expanded cell proliferation in either normoxia or hypoxia were assessed using Click-iT 5-ethynyl-2^′-deoxyuridine (EdU) Cell Proliferation Assay kit (Thermo Fisher Scientific). Cells reaching 50% confluence were incubated in the medium with 10 μM EdU at 37 °C for 18 h. Following fixation in 4% paraformaldehyde (Thermo Fisher Scientific), cells (2 × 10⁵ each group) were treated with Click-iT^® reaction cocktail for 30 min and fluorescence was evaluated by a FACS Calibur (BD Biosciences, San Jose, CA, USA) using the FCS Express software package (De Novo Software, Los Angeles, CA, USA).

Pei Y.A, & Pei M. (2021). Hypoxia Modulates Regenerative Potential of Fetal Stem Cells. Applied sciences (Basel, Switzerland), 12(1), 363.

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Quantifying Cellular Apoptosis and ROS

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To measure the reactive oxygen species (ROS), the SNU-C5/5FUR cells were stained with dihydroethidium (DHE) (Sigma) for 30 min at 37 °C, and were washed with PBS (phosphate-buffered saline) three times. To determine if the cells were apoptotic, they were stained with annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) (Sigma). Each sample was quantitatively analyzed using a CyFlow Cube 8 (Sysmex Partec, Münster, Germany). A data analysis was performed using the FCS Express software package (De Novo Software).

Yun C.W., Han Y.S, & Lee S.H. (2019). PGC-1α Controls Mitochondrial Biogenesis in Drug-Resistant Colorectal Cancer Cells by Regulating Endoplasmic Reticulum Stress. International Journal of Molecular Sciences, 20(7), 1707.

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Profiling Expanded Cell Surface Markers

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The following primary antibodies were used to detect expanded cell surface profiles: CD29 (abcam, Cambridge, MA), CD90 (BD Pharmingen, San Jose, CA), CD105 (BioLegend, San Diego, CA), the stage-specific embryonic antigen 4 (SSEA4) (BioLegend), and isotype-matched IgGs (Beckman Coulter, Fullerton, CA). The secondary antibody was goat anti-mouse IgG (H + L) R-phycoerythrin conjugated (ThermoFisher Scientific, Milford, MA). Samples (n=3) of each 2×10⁵ expanded cells were incubated on ice in cold PBS containing 0.1% ChromPure Human IgG whole molecule (Jackson ImmunoResearch Laboratories, West Grove, PA) and 1% NaN3 (Sigma-Aldrich) for 30 min. The cells were then sequentially incubated in the dark in the primary and secondary antibodies for 30 min. Fluorescence was analyzed by a FACS Calibur (BD Biosciences) using FCS Express software package (De Novo Software, Los Angeles, CA).

Pizzute T., Zhang Y., He F, & Pei M. (2016). Ascorbate-dependent impact on cell-derived matrix in modulation of stiffness and rejuvenation of infrapatellar fat derived stem cells toward chondrogenesis. Biomedical materials (Bristol, England), 11(4), 045009.

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Expanded FSDSC Immunophenotyping

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The following primary antibodies were used to detect expanded FSDSC surface immunophenotype profiles: CD29 (abcam, Cambridge, MA), CD90 (BD Pharmingen, San Jose, CA), CD105 (BioLegend, San Diego, CA), the stage-specific embryonic antigen 4 (SSEA4) (BioLegend), integrin β5 (Cell Signaling, Danvers, MA), and isotype-matched IgGs (Beckman Coulter, Fullerton, CA). The secondary antibody was goat anti-mouse IgG (H + L) R-phycoerythrin conjugated (Life Technologies). Samples (n = 3) of each 2 × 10⁵ expanded cells were incubated on ice in cold PBS containing 0.1% Chrom-Pure Human IgG whole molecule (Jackson ImmunoResearch Laboratories, West Grove, PA) and 1% NaN₃ (Sigma–Aldrich) for 30 min. The cells were then sequentially incubated in the dark in the primary and secondary antibodies for 30 min. Fluorescence was analyzed by a FACS Calibur (BD Biosciences) using FCS Express software package (De Novo Software).

Li J., He F, & Pei M. (2015). Chondrogenic priming of human fetal synovium-derived stem cells in an adult stem cell matrix microenvironment. Genes & Diseases, 2(4), 337-346.

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Evaluating Cell Proliferation in Normoxia and Hypoxia

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Pei Y.A, & Pei M. (2021). Hypoxia Modulates Regenerative Potential of Fetal Stem Cells. Applied sciences (Basel, Switzerland), 12(1), 363.

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Characterization of Adipose-Derived Stromal Cells

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Harvested cells were washed, stained with Fluorochrome-labeled antibodies and run on a FACSCANTO II (BD Biosciences, UK). Doublets were excluded using side scatter (SSC) and Forward scatter (FSC) height and width, and single cells gated for further analysis.
ADSC-derived fibroblasts were identified as CD45- single cells; differentiation of ADSC in response to each cytokine alone or in combination was determined by the expression of the lymphoid fibroblast markers podoplanin, vascular cell adhesion molecule 1 (VCAM-1, CD106) and intercellular adhesion molecule 1 (ICAM-1, CD54) relative to untreated ADSC. Results are shown as geometric mean fluorescent intensity (MFI).
All flow cytometry analysis was carried out using the FCS express software package (Version 3 Research Edition, Denovo software, USA).

Foxall R., Narang P., Glaysher B., Hub E., Teal E., Coles M.C., Ashton-Key M., Beers S.A, & Cragg M.S. (2021). Developing a 3D B Cell Lymphoma Culture System to Model Antibody Therapy. Frontiers in Immunology, 11, 605231.

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