FlexiCyte™ Assay
Fully Flexible Data Visualization and Analysis
- Advanced cell analysis of a broad range of fluorescent biomarkers and proteins
- Compliant: 21 CFR Part 11/GMP-ready
- Adjustable analysis settings before and after data acquisition
- Integrated image analysis algorithm
- Data visualization and validation during acquisition
Overview of the FlexiCyte™ Assay
The FlexiCyte™ Assay for the NucleoCounter® NC-3000™ advanced image analyzer, enables users to perform detailed cell analysis of a broad range of biomarkers and fluorescent proteins in mammalian cells. The ‘Protocol Adaptation Wizard’ guides the user through a selection of optimal settings.
FlexiCyte™ allows you to build a custom assay using a combination of LEDs from UV to far-red matched with carefully selected emission filters contained within the NucleoCounter® NC-3000™. This allows for simultaneous detection of numerous fluorescent antibodies and proteins.
The ‘Protocol Adaptation Wizard’ guides you through the selection of optimal settings. After image acquisition, data is presented as scatter plots and histograms in the Plot Manager, along with the fluorescent image. Detailed data analysis can then be performed linking individual events between images and plots for full understanding of co-localization of fluorescent events and cell (sub-)populations.
Assay Principle
Samples are stained with fluorophores specified by the user. Note that the default FlexiCyte™ protocol requires counterstaining of the sample with either DAPI or Hoechst-33342 in addition to the user specified staining. DAPI and Hoechst-33342 stain the nucleus and are used for detection (i.e. masking) of the total cell population. Hoechst-33342 stains both living and dead cells and is used for labeling samples with non-fixed cells. DAPI exclusively stains dead cells and is used for labeling cells fixed with either alcohols or aldehydes.
For staining of cellular markers, we recommend using Alexa Fluor 488, Alexa Fluor 568 and Alexa Fluor 647. These dyes are bright and photostable and have minimal spectral overlap. Avoid using dyes that are prone to photobleaching, such as FITC, PE, APC and PerCP.
For full information and recommendations, please refer to the Application Note: FlexiCyte™ Protocol for the NucleoCounter® NC-3000™ system .
Results Presented in Plot Manager
Use the Plot Manager to analyze the events acquired by the NC-View™ software with the NucleoCounter® NC-3000™. Draw polygons, quadrants, and markers in the plots to count and gate sub-populations for detailed data analysis and verification of staining events.
Instrument Solutions
Tech notes
Publications
hiPS-Mesenchymal Stem Cellss differentiation towards fibroblasts on a 3D ECM mimicking scaffold.
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Xu R, Taskin MB, Rubert M, et al. ,
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Sci Rep ,
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2015,
-
5:8480
- Instrument: NucleoCounter® NC-3000™
- Cell Type: MSCs
- Research area: Stem cell Research
Methyl gallate, gallic acid-derived compound, inhibit cell proliferation through increasing ROS production and apoptosis in hepatocellular carcinoma cells
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Huang CY, Chang YJ, Wei PL, et al. ,
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PLOS One ,
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2021,
-
16(3):e0248521
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Oncology
ROS Overproduction Sensitises Myeloma Cells to Bortezomib-Induced Apoptosis and Alleviates Tumour Microenvironment-Mediated Cell Resistance
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Caillot M, Zylbersztejn F, Maitre E, et al. ,
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Cells ,
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2020,
-
9(11):2357
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Oncology
Propyl Gallate Inhibits Hepatocellular Carcinoma Cell Growth Through the Induction of ROS and the Activation of Autophagy
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Po-Li Wei , Chien-Yu Huang , Yu-Jia Chang ,
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PLoS One ,
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2019,
-
14(1):e0210513
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Bromelain Inhibits the Ability of Colorectal Cancer Cells to Proliferate via Activation of ROS Production and Autophagy
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Tung-Cheng Chang , Po-Li Wei , Precious Takondwa Makondi , et al. ,
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PLoS One ,
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2019,
-
14(1):e0210274
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Aspirin Disrupts the Crosstalk of Angiogenic and Inflammatory Cytokines between 4T1 Breast Cancer Cells and Macrophages
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Hsieh CC, Wang CH ,
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Mediators of Inflammation ,
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2018,
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2018:6380643
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Eukaryotic Cells
- Research area: Oncology
Heat shock protein 27 influences the anti-cancer effect of curcumin in colon cancer cells through ROS production and autophagy activation
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Liang HH, Huang CY, Chou CW, et al. ,
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Life Sci ,
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2018,
-
209:43-51
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer
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Kim M, Pyo S, Kang CH, et al. ,
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PLoS One ,
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2018,
-
13(6):e0198347
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Self-assembly of tumor-targeting RNA nanoball for carrier-free delivery of therapeutic RNA
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Han S, Lee JB ,
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Journal of Industrial and Engineering Chemistry ,
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2018,
-
64,90-96
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Inhibitory effects of rosmarinic acid on pterygium epithelial cells through redox imbalance and induction of extrinsic and intrinsic apoptosis
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Chen YY, Tsai CF, Tsai MC, et al. ,
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Exp Eye Res ,
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2017,
-
160:96-105
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Identification of Prognostically Relevant Chromosomal Abnormalities in Routine Diagnostics of MultipleMyeloma Using Genomic Profiling.
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Kjeldsen E ,
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Cancer Genomics Proteomics ,
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2016,
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13(2):91-127
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Leukocytes
- Research area: Oncology
Dual targeting NG2 and GD3A using Mab-Zap immunotoxin results in reduced glioma cell viability in vitro.
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Higgins SC, Fillmore HL, Ashkan K, et al. ,
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Anticancer Res. ,
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2015,
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35(1):77-84.
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Guanidine modified polyethyleneimine-g-polyethylene glycol nanocarriers for long interfering RNA (liRNA) based advanced anticancer therapy
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Sajeesh S, Jeong Yong Choe, Tae Yeon Leea and Dong-ki Lee. ,
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J. Mater. Chem. B ,
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2015,
-
3(2):207-216
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Colorectal cancer stem cell and chemoresistant colorectal cancer cell phenotypes and increased sensitivity toNotch pathway inhibitor.
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Huang R, Wang G, Song Y, et al. ,
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Mol Med Rep ,
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2015,
-
12(2):2417-24
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
ALPPL2 Aptamer-Mediated Targeted Delivery of 5-Fluoro-2′-Deoxyuridine to Pancreatic Cancer.
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Dua P, S S, Kim S, Lee DK. ,
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Nucleic Acid Ther. ,
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2015,
-
25(4):180-7
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Carbon dots prepared from ginger exhibiting efficient inhibition of human hepatocellular carcinoma cells
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Li CL, Chung-Mao Ou,a Chih-Ching Huang, et al. ,
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J. Mater. Chem. B ,
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2014,
-
2(28):4564-4571
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Oncology
Resveratrol Inhibits Alpha-Melanocyte-Stimulating Hormone Signaling, Viability, and Invasiveness in Melanoma Cells
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Chen YJ, Chen YY, Lin YF, et al. ,
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Evid Based Complement Alternat Med. ,
-
2013,
-
2013:632121
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Alkaline Phosphatase ALPPL-2 Is a Novel Pancreatic Carcinoma-Associated Protein
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Dua P, Kang HS, Hong SM, et al. ,
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Cancer Res. ,
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2013,
-
73(6):1934-45
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Sunitinib and SU11652 inhibit acid sphingomyelinase, destabilize lysosomes and inhibit multidrug resistance
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Ellegaard AM, Groth-Pedersen L, Oorschot V, et al. ,
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Mol Cancer Ther. ,
-
2013,
-
12(10):2018-30
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Oncology
Oxidative stress is linked to ERK1/2-p16 signaling-mediated growth defect in ATM-deficient astrocytes.
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Kim J, Wong PK ,
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J Biol Chem ,
-
2009,
-
284(21)
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Neurobiology
Bumetanide attenuates acute lung injury by suppressing macrophage activation
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Hung CM, Peng CK, Wu CP, Huang KL ,
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Biochem Pharmacol ,
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2018,
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156:60-67
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Leukocytes
- Research area: Immunology
Gene Knockdown in Human Rhinovirus 1B Using 2′-OMe-modified siRNAs Results in the Reactivation of theInterferon Response
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Xie GC, Zhang Q, Pang LL, et al. ,
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Biomed Environ Sci ,
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2016,
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29(2):137-42
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Immunology
Bisphenol A Diglycidyl Ether (BADGE) and Bisphenol Analogs, but Not Bisphenol A (BPA), Activate the CatSper Ca2+ Channel in Human Sperm
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Rehfeld A, Andersson AM, Skakkebæk NE ,
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Frontiers in Endocrinology ,
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2020,
-
11:324
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Sperm Cells
- Research area: Cell Biology
Tetrahydrocarbazoles are a novel class of potent P-type ATPase inhibitors with antifungalactivity
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Bublitz M, Kjellerup L, Cohrt KO, et al. ,
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PLoS One ,
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2018,
-
13(1):e0188620
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Yeast Cells
- Research area: Cell Biology
Viable acrosome-intact human spermatozoa in the ejaculate as a marker of semen quality and fertility status
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Egeberg Palme DL, Rehfeld A, Bang AK, et al. ,
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Hum Reprod ,
-
2018,
-
33(3):361-371
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Sperm Cells
- Research area: Cell Biology
Size-Controllable Enzymatic Synthesis of Short Hairpin RNA Nanoparticles by Controlling the Rate of RNA Polymerization
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Kim H, Kim D, Jeong J, et al. ,
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Polymers ,
-
2018,
-
10(6), 589
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
Evidence of oxidative stress and mitochondrial dysfunction in spinocerebellar ataxia type 2 (SCA2) patient fibroblasts: Effect of coenzyme Q10 supplementation on these parameters.
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Cornelius N, Wardman JH, Hargreaves IP, et al. ,
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Mitochondrion ,
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2017,
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34:103-114
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
Application of image cytometry to characterize heterologous lipid flippases in yeast.
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Jensen MS, Costa SR, Theorin L, et al. ,
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Cytometry A ,
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2016,
-
;89(7):673-80
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Yeast Cells
- Research area: Cell Biology
Poly-sgRNA/siRNA ribonucleoprotein nanoparticles for targeted gene disruption.
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Ha JS, Lee JS, Jeong J, et al. ,
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J Control Release ,
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2017,
-
250:27-35
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
Synthesis of multi-functional DNA nanosphere barcode system for direct cell detection
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H Sangwoo, Jae Sung Lee and Jong Bum Lee ,
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Nanoscale ,
-
2017,
-
9(37):14094-14102
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
Generation of siRNA Nanosheets for Efficient RNA Interference
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Kim H, Lee JS, Lee JB ,
-
Sci Rep ,
-
2016,
-
6:25146
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
Cell-Based Lipid Flippase Assay Employing Fluorescent Lipid Derivatives.
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Jensen MS, Costa S, Günther-Pomorski T, López-Marqués RL. ,
-
Methods Mol Biol. ,
-
2016,
-
1377:371-82.
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Yeast Cells
- Research area: Cell Biology
Self-assembled DNA-Guided RNA Nanovector via Step-wise Dual Enzyme Polymerization (SDEP) for Carrier-free siRNA Delivery
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Park Y, Hyejin Kim, and Jong Bum Lee ,
-
ACS Biomater. Sci. Eng ,
-
2016,
-
2(4):616-624
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
Application of an Image Cytometry Protocol for Cellular and Mitochondrial Phenotyping on Fibroblasts from Patients with Inherited Disorders
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Fernandez-Guerra P, Lund M, Corydon TJ, et al. ,
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JIMD Rep ,
-
2016,
-
27:17-26
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
Hyperphosphatemia induces protective autophagy in endothelial cells through the inhibition of Akt/mTOR signaling
-
Hsu YJ, Hsu SC, Huang SM, et al. ,
-
J Vasc Surg ,
-
2015,
-
62(1):210-221
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Research area: Cell Biology
Comparison of three thiol probes for determination of apoptosis-related changes in cellular redox status.
-
Skindersoe ME, Kjaerulff S. ,
-
Cytometry A ,
-
2014,
-
85(2):179-87
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
Long dsRNA-mediated RNA Interference and Immunostimulation: A Targeted Delivery Approach using Polyethyleneimine based Nano-Carriers
-
Sajeesh S, Lee TY, Hong SW, et al. ,
-
Mol Pharm ,
-
2014,
-
11(3):872-84
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
RNA Interference-Mediated Simultaneous Silencing of Four Genes Using Cross-Shaped RNA
-
Lee TY, Chang CI, Lee D, et al. ,
-
Mol Cells ,
-
2013,
-
35(4):320-6. doi: 10.1007/s10059-013-2316-7
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
Efficient intracellular delivery and multiple-target gene silencing triggered by tripodal RNA based nanoparticles: a promising approach in liver-specific RNAi delivery.
-
Sajeesh S, Lee TY, Kim JK, et al. ,
-
J Control Release ,
-
2014,
-
196:28-36
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Research area: Cell Biology
Secondary Coenzyme Q10 deficiency and oxidative stress in cultured fibroblasts from patients with Riboflavin Responsive Multiple Acyl-CoA Dehydrogenation Deficiency
-
Cornelius N, Byron C, Hargreaves I, et al. ,
-
Hum Mol Genet ,
-
2013,
-
22(19):3819-27
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
RNF8 and RNF168 but not HERC2 are required for DNA damage-induced ubiquitylation in chicken DT40 cells
-
Oestergaard VH, Pentzold C, Pedersen RT, et al. ,
-
DNA Repair ,
-
2012,
-
11(11):892-905
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Research area: Cell Biology
The calcium-sensing receptor is essential for calcium and bicarbonate sensitivity in human spermatozoa
-
Boisen IM, Rehfeld A, Mos I, et al. ,
-
The Journal of Clinical Endocrinology and Metabolism ,
-
2020,
-
dgaa937
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Sperm Cells
- Research area: Animal Reproduction