In cell analyzer 2200

Manufactured by GE Healthcare

Sourced in United Kingdom, United States, Japan

The IN Cell Analyzer 2200 is a high-content screening system designed for cellular imaging and analysis. It provides automated cellular image acquisition and analysis capabilities for a wide range of applications in life science research.

Automatically generated - may contain errors

Lab products found in correlation

150 protocols using in cell analyzer 2200

Immunofluorescent Assay of Osteopontin Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The epithelial type of the resulting cultured cells was confirmed by an immunofluorescent assay of osteopontin (OPN) expression [36 (link)]. The groups were as follows: a control, 200 μM CoCl₂ added to the incubation medium, and 200 μM CoCl₂+1 μM B[a]P added to the incubation medium.
For immunocytochemical analysis, the culture medium was discarded, and the cells were rinsed with PBS and fixed with 4% paraformaldehyde for 10 min at room temperature. Permeabilization was carried out by means of a Triton X-100 solution, and 1% BSA served as a blocker. Then, the cells were incubated with an anti-OPN rabbit polyclonal antibody (Abcam cat. # ab844) (primary antibody, dilution 1 : 200) for 1 h at room temperature. After that, a secondary fluorescently labeled antibody (rabbit anti-human IgG (H+L) cross-adsorbed secondary antibody, DyLight 550, Invitrogen, 1 : 1000) was used. The incubation lasted for 30 min at room temperature. Phase contrast and fluorescent images of cells were assessed on IN Cell Analyzer 2200 (GE Healthcare, UK) in bright-field mode and in DAPI and Cy3 channels. The experiments were performed in triplicate.

Perepechaeva M.L., Klyushova L.S, & Grishanova A.Y. (2023). AhR and HIF-1α Signaling Pathways in Benign Meningioma under Hypoxia. International Journal of Cell Biology, 2023, 6840271.

+ Open protocol

+ Expand

Cell Viability and Death Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell membrane integrity and cell death induction were evaluated using propidium iodide (PI) and Hoechst 33,342 staining. After the differentiation protocol and 6-OHDA and rotenone incubations, cell nuclei were stained with 4 µM PI (cat. P21493, Thermo Fisher Scientific, Hampton, NH, USA) and 1 µg/mL of Hoechst 33342 (cat. B2261, Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). For being a cell-impermeant stain, only cells without an intact membrane were labeled with PI, while Hoechst 33342 labeled all cell nuclei. Both PI and Hoechst 33,342 were added directly to the media without aspirating it in order to avoid removing dead, PI-positive, cells. Cells were incubated with both stains for 5 min and visualized using an INCell Analyzer 2200 (GE Healthcare, Chicago, IL, USA) cell imaging system with 10× magnification (Nikon 10×/0.45, Plan Apo, CFI/60, Tokyo, Japan). The image analysis was performed using IN Cell Analyzer 1000 analysis Software-Developer Toolbox (GE Healthcare, Chicago, IL, USA). The PI negative cells were divided by the total number of cells to calculate the live cell population. Data were normalized to the average control (100%) condition.

Simões R.F., Oliveira P.J., Cunha-Oliveira T, & Pereira F.B. (2022). Evaluation of 6-Hydroxydopamine and Rotenone In Vitro Neurotoxicity on Differentiated SH-SY5Y Cells Using Applied Computational Statistics. International Journal of Molecular Sciences, 23(6), 3009.

+ Open protocol

+ Expand

Immunocytochemistry of Pluripotent Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Patient-derived iPSCs were fixed for 15 mins in 4% paraformaldehyde, washed with Dulbecco’s Phosphate-Buffered Saline (DPBS), permeabilized with 0.5% Triton X-100 in DPBS (10 mins), followed by incubation with Image-iT^™ FX signal enhancer (ThermoFisher) for 40 mins at ambient temperature in a humidified environment. Primary antibodies (SOX2, OCT4, NANOG and SSEA4) were diluted in the Image-iT^™ FX signal enhancer blocking buffer and then incubated with the cells overnight at 4 °C. Following a DPBS wash, corresponding secondary antibodies conjugated with Alexa Fluor 488 or Alex Fluor 594 were added and incubated for 1 h at ambient temperature (Table 2). Cells were stained with Hoechst 33342 (15 mins), washed, and imaged using an INCell Analyzer 2200 (GE Healthcare) and a 20× objective lens with Texas Red, FITC and DAPI filter sets.

Hong J., Xu M., Li R., Cheng Y.S., Kouznetsova J., Beers J., Liu C., Zou J, & Zheng W. (2019). Generation of an induced pluripotent stem cell line (TRNDi008-A) from a Hunter syndrome patient carrying a hemizygous 208insC mutation in the IDS gene. Stem cell research, 37, 101451.

+ Open protocol

+ Expand

Quantifying Lipid Droplets in Breast Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MDA-MB-231 and MCF-7 cells were seeded at 2 × 10⁴ cells in 96 microwell plates and treated or not with 4-cholesten-3-one for 24 h. Cells were washed with PBS and fixed with 4% cold paraformaldehyde, then stained with Nile red (5 μg/mL) prepared in PBS from a stock solution at 0.5 mg/mL in DMSO. After incubation (30 min in the dark), the cells were washed with 1X PBS and then the cell nucleus was counter-stained (1:200) from a stock solution of DAPI 5 mg/mL diluted in PBS. Cells were washed and mounted using an Ibidi aqueous mounting medium. Fluorescence microscopy was performed using the IN Cell Analyzer 2200 (GE Healthcare, Vélizy-Villacoublay, France) with the Nikon focal lens (20X / 0.45). Nile red stains the lipid droplets and is observed using the red color channel (excitation at 542 nm, emission at 597 nm with an exposure time of 100 ms), whereas DAPI is observed in the blue color channel (excitation at 390, emission at 435 nm with an exposure time of 500 ms). Images were acquired with both detection channels and merged using ImageJ software.

Elia J., Carbonnelle D., Logé C., Ory L., Huvelin J.M., Tannoury M., Diab-Assaf M., Petit K, & Nazih H. (2019). 4-cholesten-3-one decreases breast cancer cell viability and alters membrane raft-localized EGFR expression by reducing lipogenesis and enhancing LXR-dependent cholesterol transporters. Lipids in Health and Disease, 18, 168.

+ Open protocol

+ Expand

Immunofluorescence Staining of iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunofluorescence staining, patient iPSCs were fixed with 4% paraformaldehyde for 30 min at RT, permeabilized with 0.3% Triton X-100 in Dulbecco’s phosphate-buffered saline (DBPS) for 15 min, and washed with DPBS. Cells were blocked using Image-iT™ FX signal enhancer (Thermo Fisher Scientific) for 1 h and incubated with primary antibodies, including SOX2, OCT4, NANOG and SSEA4, overnight at 4 °C. Cells were then washed and incubated with corresponding secondary antibody conjugated with Alexa Fluor 488 or Alexa Fluor 594 for 1 h at room temperature (antibodies used are listed in Table 2). Cells were washed and stained with Hoechst 33342 for 15 min and imaged using an INCell Analyzer 2200 imaging system (GE Healthcare) with 20× objective lens and Texas Red, FITC and DAPI filter sets.

Baskfield A., Li R., Beers J., Zou J., Liu C, & Zheng W. (2019). Generation of an induced pluripotent stem cell line (TRNDi004-I) from a Niemann-Pick disease type B patient carrying a heterozygous mutation of p.L43_A44delLA in the SMPD1 gene. Stem cell research, 37, 101436.

+ Open protocol

+ Expand

Clonogenic Assay for Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Stably or transiently transfected PC3 cells were plated in 6 well plates (200 cells/well) and incubated for 10 days in complete medium. HeLa cells were transfected, detached 24 h post transfection, plated in 6 well plates (100, 200 or 400 cells/well) and incubated for 7 days in complete medium. Where indicated, medium was supplemented with the indicated concentrations of docetaxel or DMSO. Colonies were then fixed with 4% paraformaldehyde (PFA) and stained with crystal violet (0.01% in PBS). Images were acquired using a bright field microscope. Alternatively, fixed colonies were incubated with HCS CellMask™ Deep Red (cat number: H32721, Thermo Fisher Scientific) and 4′,6-diamidino-2-phenylindole (DAPI, cat number: D1306, Thermo Fisher Scientific) and images were acquired and analysed using IN Cell Analyzer 2200 (GE Healthcare Life Sciences).

Cisse O., Quraishi M., Gulluni F., Guffanti F., Mavrommati I., Suthanthirakumaran M., Oh L.C., Schlatter J.N., Sarvananthan A., Broggini M., Hirsch E., Falasca M, & Maffucci T. (2019). Downregulation of class II phosphoinositide 3-kinase PI3K-C2β delays cell division and potentiates the effect of docetaxel on cancer cell growth. Journal of Experimental & Clinical Cancer Research : CR, 38, 472.

+ Open protocol

+ Expand

Conditional Immortalized Podocyte Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols

Kidneys were isolated from GSK3α^fl/fl mice and used to make a temperature‐sensitive SV40 conditionally immortalized podocyte cell line as described previously.19 Cells were cultured at 33°C and when 50% confluent were transduced with a lentivirus expressing Cre recombinase as described previously.10 Transduction was in RPMI media with hexadimethrine bromide (Sigma) at 4 μg/mL and the virus used at a multiplicity of infection of 1. Following a 24‐hour incubation, the lentivirus was removed and replaced with fresh media. Cells were thermo‐switched to 38°C and incubated for a further 7 days before imaging and protein extraction.
To determine ciGSK3αKO cell number, cells were washed three times in PBS, the nuclei stained with Hoechst (Sigma) at 1 μg/mL and imaged using an IN Cell analyzer 2200 (GE Healthcare) and data analyzed using IN Cell analyzer Developer software (GE Healthcare).

Hurcombe J.A., Lay A.C., Ni L., Barrington A.F., Woodgett J.R., Quaggin S.E., Welsh G.I, & Coward R.J. (2019). Podocyte GSK3α is important for autophagy and its loss detrimental for glomerular function. FASEB bioAdvances, 1(8), 498-510.

+ Open protocol

+ Expand

Immunostaining of Neuronal Cultures

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary neuron or cell line cultures were fixed with 4% paraformaldehyde, 4% sucrose in phosphate-buffered saline and washed five times in PBS. Immunostaining of neuronal cultures was carried out as described previously (Henderson et al., 2017 ; Henderson et al., 2018 (link)). Cells were permeabilized in 3% BSA + 0.3% TX-100 in PBS for 15 minutes at room temperature. After a PBS wash, cells were blocked for 50 minutes with 3% BSA in PBS prior to incubation with primary antibodies for 2 hours at room temperature. Primary antibodies used were targeting pS129 α-synuclein (81A, CNDR, 1:5,000) and NeuN (Millipore Cat#MAB377, 1:1,500). Cells were washed 5x with PBS and incubated with secondary antibodies for 1 hour at room temperature. After 5x wash with PBS, cells were incubated in DAPI (ThermoFisher Cat#D21490, 1:10,000) in PBS. 96-well plates were imaged on In Cell Analyzer 2200 (GE Healthcare) and analyzed in the accompanying software. A standard intensity-based threshold was applied to the pS129 α-synuclein channel equally across plates and positive area was quantified. For NeuN quantification, an object-based analysis was applied to identify objects of specified size and intensity. All quantification was optimized and applied equally across all conditions.

Henderson M.X., Covell D.J., Chung C.H., Pitkin R.M., Sandler R.M., Decker S.C., Riddle D.M., Zhang B., Gathagan R.J., James M.J., Trojanowski J.Q., Brunden K.R., Lee V.M, & Luk K.C. (2019). Characterization of novel conformation-selective α-synuclein antibodies as potential immunotherapeutic agents for Parkinson’s disease. Neurobiology of disease, 136, 104712.

+ Open protocol

+ Expand

High Content Imaging of Cell Morphology

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell morphology was assessed by High Content Imaging using the InCell Analyzer 2200 and IN Cell Investigator image analysis software (GE Healthcare). Morphological parameters were assessed in the nuclear and cytoplasmic compartments which were labelled with the fluorescent nuclear and cytoskeletal stains, Hoechst and Phalloidin respectively. Measured parameters included: nuclear area, nuclear intensity, nuclear displacement, cell area, nuclear to cell ratio, cell spread, cell roundness and cell intensity/area.

Harkness L., Chen X., Gillard M., Gray P.P, & Davies A.M. (2019). Media composition modulates human embryonic stem cell morphology and may influence preferential lineage differentiation potential. PLoS ONE, 14(3), e0213678.

+ Open protocol

+ Expand

Fluorescence-based Unscheduled DNA Synthesis (UDS)

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescent-based UDS was performed as follows. Cells seeded on coverslips were arrested at M phase by 50 nM nocodazole for 15 h, released by washing with PBS and then cultured for 3 h. Cells were then ultraviolet irradiated with 0 or 15 J m⁻² and incubated for 3 h in medium containing 5-ethynyl-20-deoxyuridine (EdU; Invitrogen), followed by washing with PBS and fixation with 4% formaldehyde. Cells were permeabilized with 0.5% Triton in PBS, and EdU incorporation and total DNA were visualized using Click-it Alexa Fluor 594 and Hoechst 33342, respectively, according to the manufacturer’s instructions (Invitrogen). The total DNA content of each cell was quantified by measuring Hoechst 33342. The specific repair capacity of G1 cells was quantified in 1,500 cells by determining the overall nuclear fluorescence of EdU using the IN CELL Analyzer 2200 (GE Healthcare Life Sciences). The fluorescence values were normalized to the fluorescence in shCtl cells, which was set at 100%.

Niida H., Matsunuma R., Horiguchi R., Uchida C., Nakazawa Y., Motegi A., Nishimoto K., Sakai S., Ohhata T., Kitagawa K., Moriwaki S., Nishitani H., Ui A., Ogi T, & Kitagawa M. (2017). Phosphorylated HBO1 at UV irradiated sites is essential for nucleotide excision repair. Nature Communications, 8, 16102.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!