Spot imaging software

Manufactured by Nikon

Sourced in Japan

SPOT imaging software is a digital imaging platform designed for microscopy applications. It provides tools for image capture, processing, and analysis. The software is compatible with various Nikon microscopes and digital cameras to facilitate seamless integration and workflow.

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Lab products found in correlation

24 well biocoat matrigel invasion chambers, by BD (5 mentions) Biocoat matrigel invasion chamber, by BD (3 mentions) Crystal violet, by Merck Group (3 mentions) Millicell hanging insert, by Merck Group (3 mentions) 24 well millicell hanging insert, by Merck Group (2 mentions) Metamorph software, by Molecular Devices (2 mentions) Prism software, by GraphPad (2 mentions) Inverted microscope, by Nikon (2 mentions) 24 well milli cell hanging insert, by BD (2 mentions) Colorimetric assay, by Sekisui (2 mentions) Porous membrane, by BD (1 mentions) 24 well migration chamber, by Corning (1 mentions) Transwell chamber, by Corning (1 mentions) Nuclear fast red solution, by Vector Laboratories (1 mentions) Hemavet blood analyzer, by Drew Scientific (1 mentions) Eclipse e600 microscope, by Nikon (1 mentions)

18 protocols using spot imaging software

Cell Migration and Invasion Assay

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Migration and invasion assays were performed in 24-well millicell hanging insert (Millipore) or 24-well BioCoat Matrigel Invasion Chambers (BD Biosciences). In brief, 1 × 10⁵ cells were seeded to the top chamber and 10% FBS in medium was added to the bottom chamber as a chemoattractant. After 24 or 48-h incubation, the number of cells that invaded through the membrane (migration) or Matrigel (invasion) was counted in 10 fields and imaged using SPOT imaging software (Nikon).

Ming X.Y., Fu L., Zhang L.Y., Qin Y.R., Cao T.T., Chan K.W., Ma S., Xie D, & Guan X.Y. (2016). Integrin α7 is a functional cancer stem cell surface marker in oesophageal squamous cell carcinoma. Nature Communications, 7, 13568.

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Quantification of Cell Migration and Invasion

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For detection of migration and invasion of

α

δ

^{+}

cells, 5

\times

^{4}

cells were added onto a porous membrane (pore size, 8

μ

m; BD Biosciences) that was coated with 2 mg/mL Matrigel. After 48-hour incubation at 37

^{\circ}

C, cells were washed three times with PBS, and fixed with 4% neutral formaldehyde prior to routine hematoxylin staining for 5 min. The number of cells that invaded through the membrane (migration) or Matrigel (invasion) was counted in 10 representative fields at

\times

20 magnification. Images were acquired and analyzed using SPOT imaging software (Nikon).

Huang C., Li Y., Zhao W., Zhang A., Lu C., Wang Z, & Liu L. (2019). 21 may be a potential marker for cancer stem cell in laryngeal squamous cell carcinoma. Cancer Biomarkers, 24(1), 97-107.

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Invasion and Migration Assay Protocol

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Invasion and migration assays were conducted in 24-well BioCoat Matrigel Invasion Chambers (BD Biosciences) or 24-well Millicell hanging inserts (Millipore). Cells re-suspended in serum free DMEM were added to the top chamber and DMEM supplemented with 10% FBS was added to the bottom chamber as a chemoattractant. After 48 hrs incubation at 37°C, the number of cells that invaded through the Matrigel (invasion) or membrane (migration) was counted in 10 fields under 4x objective lens and imaged using SPOT imaging software (Nikon, Japan).

Chai S., Tong M., Ng K.Y., Kwan P.S., Chan Y.P., Fung T.M., Lee T.K., Wong N., Xie D., Yuan Y.F., Guan X.Y, & Ma S. (2014). Regulatory role of miR-142-3p on the functional hepatic cancer stem cell marker CD133. Oncotarget, 5(14), 5725-5735.

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Cell Migration and Invasion Assay

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Cell ability of migration and invasion were performed using 24-well migration chamber (Corning, NY) or 24-well BioCoat Matrigel Invasion Chambers (BD Biosciences, CA) with polyethylene terephthalate (PET)-based membrane of 8.0-um pore size. In Brief, 1x10⁵ cells were suspended with serum-free culture medium and seeded in the top chambers. Complete culture medium with 10% FBS was used in the bottom chambers. The chambers were collected after 24/48 hours’ incubation. Cells which moved through the membrane were fixed with 4% PFA and stained with 0.5% crystal violet solution. The number of cells invaded through the membrane per field were counted and imaged using SPOT imaging software (Nikon) under microscope.

Cao T.T., Xiang D., Liu B.L., Huang T.X., Tan B.B., Zeng C.M., Wang Z.Y., Ming X.Y., Zhang L.Y., Jin G., Li F., Wu J.L., Guan X.Y., Lu D, & Fu L. (2017). FZD7 is a novel prognostic marker and promotes tumor metastasis via WNT and EMT signaling pathways in esophageal squamous cell carcinoma. Oncotarget, 8(39), 65957-65968.

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Characterization of iPSC-Derived Neurons

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Data for each iPSC line are from two to six independent experiments with three technical replicates within each experiment. Within each immunocytochemistry-based experiment, at least five images were taken on each of at least three different fluorescently labeled coverslips per time point per line using a Nikon inverted microscope and Spot imaging software. The images were analyzed for antigen specificity using MetaMorph Software (Molecular Devices). Calcium imaging data were collected from a minimum of three coverslips per time point from each line. A minimum of 25 neurons were recorded from each coverslip. In all experiments, the evaluator was blinded to the cell line and treatment condition. Data were statistically analyzed with Prism software (GraphPad) first for normality using the D’Agostino-Pearson omnibus normality test, followed by the parametric one-way ANOVA and Tukey’s multiple comparison test of significance or the non-parametric Kruskal-Wallis test and Dunn’s multiple comparison test of significance; α = 0.05. Student’s t test was used when appropriate. Data are presented as the average ± SEM with the statistical test used indicated in each figure legend.

Schwab A.J, & Ebert A.D. (2015). Neurite Aggregation and Calcium Dysfunction in iPSC-Derived Sensory Neurons with Parkinson’s Disease-Related LRRK2 G2019S Mutation. Stem Cell Reports, 5(6), 1039-1052.

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Transwell Migration and Matrigel Invasion Assays

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For transwell migration assays, 1 × 10⁵ cells were added to the top non-coated chamber (24-well insert; pore size, 8μm; BD Biosciences). For Matrigel invasion assays, 2 × 10⁵ cells were plated in the top chamber with Matrigel-coated membrane (24-well insert; pore size, 8 μm; BD Biosciences). In both assays, cells were plated in RPM1640 medium without serum or growth factors, and RPM1640 medium supplemented with 10% FBS was used as a chemoattractant in the lower chamber. The cells were incubated for 18h and cells that did not migrate or invade through the pores were removed by a cotton swab. Cells on the lower surface of the membrane were fixed and stained with crystal violet (Sigma-Aldrich). The number of cells was counted in ten fields under a ×20 objective lens and imaged using SPOT imaging software (Nikon, Japan).

Wang Z., Xie J., Yan M., Wang J., Wang X., Zhang J., Zhang Y., Li P., Lei X., Huang Q., Lin S., Guo X, & Liu Q. (2016). Downregulation of ATOH8 induced by EBV-encoded LMP1 contributes to the malignant phenotype of nasopharyngeal carcinoma. Oncotarget, 7(18), 26765-26779.

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Cell Migration and Invasion Assay

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Migration and invasion assays were performed in 24-well millicell hanging insert (Millipore) or 24-well BioCoat Matrigel Invasion Chambers (BD Biosciences). In brief, 1×10⁵ cells were seeded to the top chamber and 10% FBS in medium was added to the bottom chamber as a chemoattractant. After 24 h or 48 h incubation, the number of cells that invaded through the membrane (migration) or Matrigel (invasion) was counted in 10 fields and imaged using SPOT imaging software (Nikon).

Ming X.Y., Zhang X., Cao T.T., Zhang L.Y., Qi J.L., Kam N.W., Tang X.M., Cui Y.Z., Zhang B.Z., Li Y., Qin Y.R, & Guan X.Y. (2018). RHCG Suppresses Tumorigenicity and Metastasis in Esophageal Squamous Cell Carcinoma via Inhibiting NF-κB Signaling and MMP1 Expression. Theranostics, 8(1), 185-198.

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Fluorescence Imaging Quantifies Antigen Expression

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At least five images were taken on each of at least three different fluorescently labeled coverslips per timepoint using a Nikon inverted microscope and Spot imaging software. All experiments were repeated at least 2 times. The images were analyzed for antigen specificity using MetaMorph Software (Molecular Devices Inc.). Data were statistically analyzed via one-way ANOVA or Student’s t-test as appropriate with Prism software, α = 0.05 (GraphPad). Data are presented as the average ± S.E.M. using two independent control lines and two independent SMA lines.

Schwab A.J, & Ebert A.D. (2014). Sensory Neurons Do Not Induce Motor Neuron Loss in a Human Stem Cell Model of Spinal Muscular Atrophy. PLoS ONE, 9(7), e103112.

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Cell Migration and Invasion Assay

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Migration and invasion assays were performed in 24-well milli-cell hanging insert (BD Biosciences) or 24-well BioCoat Matrigel Invasion Chambers (BD Biosciences). In brief, 5 × 10⁴ cells were seeded to the top chamber and 10% FBS in medium was added to the bottom chamber as a chemoattractant. After 24 or 48 h incubation, the number of cells that invaded through the membrane (migration) or Matrigel (invasion) was counted in 10 fields and imaged using SPOT imaging software (Nikon).

Jia Y., Yan Q., Zheng Y., Li L., Zhang B., Chang Z., Wang Z., Tang H., Qin Y, & Guan X.Y. (2022). Long non-coding RNA NEAT1 mediated RPRD1B stability facilitates fatty acid metabolism and lymph node metastasis via c-Jun/c-Fos/SREBP1 axis in gastric cancer. Journal of Experimental & Clinical Cancer Research : CR, 41, 287.

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Transwell Cell Migration Assay

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Cell migration assay was conducted in 24‐well Transwell chambers (Corning Costar Corporation, Corning, NY, USA). After 24-hour transfection, cells were suspended in 0.5 ml serum-free medium and loaded on the top chamber, and medium supplemented with 10% FBS was added to the bottom chamber as the chemoattractant. After 36 hours of incubation at 37 °C, cells that had migrated through the membrane were fixed with formaldehyde and stained with 1% crystal violet for 30 minutes. The migrated cells attached to the lower surface of the membrane insert were counted in five random fields under a 20× objective lens and imaged using SPOT imaging software (Nikon, Tokyo, Japan). Three independent experiments were performed.

Liang Z., Chen Y., Zhao Y., Xu C., Zhang A., Zhang Q., Wang D., He J., Hua W, & Duan P. (2017). miR-200c suppresses endometriosis by targeting MALAT1 in vitro and in vivo. Stem Cell Research & Therapy, 8, 251.

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