C1 confocal imaging system

Manufactured by Nikon

Sourced in Japan, United States

The C1 confocal imaging system is a laboratory equipment product offered by Nikon. It is designed to perform confocal microscopy, a technique that allows for high-resolution imaging of biological samples. The core function of the C1 system is to capture detailed, three-dimensional images of specimens by using a focused laser beam to scan the sample and collect the emitted fluorescence signals.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (3 mentions) Supermount, by BioGenex (2 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (2 mentions) Flexstation 3, by Molecular Devices (1 mentions) Fitc conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Cm3050s cryostat, by Leica (1 mentions) A1r confocal imaging system, by Nikon (1 mentions) Alexa fluor 594 conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Cellmatrix type 1 p, by Nitta Gelatin (1 mentions) Alexa fluor 594 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 546 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions) L9393, by Merck Group (1 mentions) Imaris software, by Oxford Instruments (1 mentions) Cellrox green reagent, by Thermo Fisher Scientific (1 mentions) Te2000 microscope, by Nikon (1 mentions)

9 protocols using c1 confocal imaging system

Rhodamine-labeled Protocell Uptake Evaluation

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Uptake of rhodamine-labeled protocells was measured in vitro using: (1) A FlexStation 3 microplate reader system (Molecular Devices, Sunnyvale, CA) in order to evaluate the effects of varying treatment concentrations and incubation times, (2) a Nikon C1 confocal imaging system (Nikon Instruments Inc., Melville, NY) to evaluate uptake in differentiated motoneuron-like NSC-34 cells as well as in L6 muscle cells to assess targeting efficiency and specificity and (3) transmission electron microscopy (TEM) of motoneuron-like NSC-34 cells after incubation with CTB-modified protocells to assess targeted cell binding and internalization.

Gonzalez Porras M.A., Durfee P.N., Gregory A.M., Sieck G.C., Brinker C.J, & Mantilla C.B. (2016). A novel approach for targeted delivery to motoneurons using cholera toxin-B modified protocells. Journal of neuroscience methods, 273, 160-174.

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Immunohistochemical Analysis of Ischemia-Reperfusion

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Rats were deeply anesthetized and transcardially perfused with normal saline solution, followed by 4% paraformaldehyde in 0.1 M PBS 24-h after ischemia-reperfusion. The brains were removed and post-fixed in 4% paraformaldehyde for 4-h, then transferred into 30% sucrose solution, until they sank to the bottom of the container. Coronal sections (20 µm) were made using a Leica CM3050S cryostat (Leica Microsystems, Wetzlar, Germany). Sections were blocked with 3% normal goat serum (diluted in PBS containing 0.3% Triton X-100) for 1-h and incubated with primary antibodies (anti-MAP2 and anti-GFAP, 1:1000, Chemicon) overnight at 4 °C. After rinsing with PBS, sections were incubated with rhodamine-conjugated goat anti-rabbit IgG (for MAP2, Millipore) and FITC-conjugated goat anti-mouse IgG (for GFAP, Invitrogen) as secondary antibodies (1:200) for 1-h. Fluorescent images were captured by a Nikon C-1 confocal imaging system (Nikon, Japan).

Wu X.M., Qian C., Zhou Y.F., Yan Y.C., Luo Q.Q., Yung W.H., Zhang F.L., Jiang L.R., Qian Z.M, & Ke Y. (2017). Bi-directionally protective communication between neurons and astrocytes under ischemia. Redox Biology, 13, 20-31.

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Immunofluorescence Staining of Cellular Proteins

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Cells were fixed with 1% or 2% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min for PP-MRLC or other proteins, respectively. The samples were permeabilized by incubation in 0.5% Triton-X100 in PBS for 10 min and blocked with 0.5% skim milk (Megmilk Snow Brand Co., Ltd., Hokkaido, Japan) in PBS for PP-MRLC or with 0.5% bovine serum albumin (Sigma-Aldrich) in PBS for other proteins. All samples were incubated with primary antibody overnight at 4°C for nuclei or at room temperature for other proteins. The appropriate secondary antibody and Alexa Fluor-488-conjugated phalloidin were then added for 3 h at room temperature. Cells were incubated with DAPI at 37°C for 1 h to stain nuclei. Images of fluorescent cells were captured using a confocal laser scanning microscope (C1 confocal Imaging System (Nikon) or A1R Confocal Imaging System).

Ishida S., Tanaka R., Yamaguchi N., Ogata G., Mizutani T., Kawabata K, & Haga H. (2014). Epithelial Sheet Folding Induces Lumen Formation by Madin-Darby Canine Kidney Cells in a Collagen Gel. PLoS ONE, 9(8), e99655.

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Detecting Aggresome-like Inclusions in N27 Cells

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Aggresome-like inclusion bodies were determined using immnostaining for ubiquitin-positive aggregates as previously described (Xiong et al., 2013 (link)). Sub-confluent N27 cells were grown on glass coverslips in 6-well plates and treated with quinones for the indicated times. Cells were washed three times with PBS and fixed in 3.7% (v/v) formaldehyde in PBS for 12min, and then permeabilized with 0.1% (v/v) Triton X-100 in PBS for 10min. After extensively washing with PBS, cells were incubated in blocking buffer (RPMI-1640 containing 10% FBS) for 1h, and then incubated with anti-ubiquitin antibody (1:100) overnight at 4°C. Cells were then washed three times in TBST and incubated with Alexa Fluor 594 conjugated goat anti-rabbit IgG (1:1000, Jackson Immunoresearch Labs) containing DAPI (1µg/ml) for 30min. Coverslips were washed three times with TBST then rinsed in distilled water, inverted and mounted on glass slides using SuperMount (BioGenex, USA). Cells were viewed on a Nikon TE2000 microscope with a Nikon C1 confocal imaging system.

Xiong R., Siegel D, & Ross D. (2014). Quinone-Induced Protein Handling Changes: Implications for Major Protein Handling Systems in Quinone-Mediated Toxicity. Toxicology and applied pharmacology, 280(2), 285-295.

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Collagen Overlay Assay for Cell Culture

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A 1.6 mg/ml collagen type I gel (Cellmatrix type I-P; Nitta Gelatin, Osaka, Japan) was used for the collagen overlay assay (Ishida et al., 2014 (link)). An 8.0-mm-radius glass dish was filled with 150 μl of the collagen gel, onto which trypsinized cells (2.0 × 10³) were seeded. After culture for 3 or 5 d, a collagen gel solution (75 μl) was poured onto the cells, and the sample was incubated for 30 min at 37°C to induce gelation. The dish was then filled with culture medium. After 1 d, cells were fixed with 2% paraformaldehyde in PBS for 10 min, permeabilized with 0.5% Triton X-100 in PBS for 5 min, and blocked with 0.5% bovine serum albumin (Sigma-Aldrich) in PBS for 1 h. Samples were incubated with Alexa Fluor 555–phalloidin (1:300 dilution in PBS) overnight at room temperature. Fluorescence images were obtained using a confocal laser-scanning microscope (C1 confocal imaging system; Nikon) with a 60× objective.

Sakane A., Yoshizawa S., Nishimura M., Tsuchiya Y., Matsushita N., Miyake K., Horikawa K., Imoto I., Mizuguchi C., Saito H., Ueno T., Matsushita S., Haga H., Deguchi S., Mizuguchi K., Yokota H, & Sasaki T. (2016). Conformational plasticity of JRAB/MICAL-L2 provides “law and order” in collective cell migration. Molecular Biology of the Cell, 27(20), 3095-3108.

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Visualizing 3D Cellular Morphology

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Cells were fixed with 1% or 2% formaldehyde in phosphate-buffered saline (PBS) for 5 min, permeabilized with 0.5% Triton-X100 in PBS for 5 min, and blocked with 0.5% bovine serum albumin (Sigma) or 0.5% skim milk (Megmilk Snow Brand Co., Ltd., Sapporo, Japan) in PBS. Reaction with the primary antibody was performed at room temperature overnight. The primary antibodies used were L9393 (Sigma) for laminin-111, anti-phospho-MRLC (Ser 19) mouse IgG (Cell Signaling Technology Japan, K.K., Japan), and anti-phospho-MRLC (Thr18/Ser19) rabbit IgG (Cell Signaling Technology) at a dilution of 1:250, 1:200, and 1:200, respectively. All samples were incubated with a mixed solution consisting of the secondary antibody and Alexa Fluor-488 phalloidin (Invitrogen) for F-actin staining at 37 °C for 1 h. AlexaFluor-594 goat anti-rabbit IgG (H + L) and AlexaFluor-546 goat anti-mouse IgG (H + L) were used as secondary antibodies (Molecular Probes, Eugene, OR, USA) at a concentration of 10 μg/mL. Fluorescent images were captured using confocal laser scanning microscopy (C1 confocal imaging system; Nikon). We used the Imaris software (Bitplane AG, Zürich, Switzerland) for 3D reconstruction of confocal images. This software enabled the visualization of 3D morphologies of the epithelial cells.

Imai M., Furusawa K., Mizutani T., Kawabata K, & Haga H. (2015). Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate. Scientific Reports, 5, 14208.

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Confocal Imaging of Retina Sections

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Imaging of the retina sections was performed using a Nikon C1 confocal imaging system with solid-state laser excitation at 488 and 568 nm. Confocal images were stitched together using Adobe Photoshop CS2.

Ferdous S., Grossniklaus H.E., Boatright J.H, & Nickerson J.M. (2019). Characterization of LSD1 Expression Within the Murine Eye. Investigative Ophthalmology & Visual Science, 60(14), 4619-4631.

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Immunofluorescence Imaging of Cells in Collagen Gel

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Cells cultured on a glass-bottom dish covered with a collagen gel were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) and permeabilized with 0.5% Triton X-100 in PBS. Blocking was performed using 0.5% bovine serum albumin (BSA)/PBS. The cells were incubated with a specific primary antibody, and reacted with a secondary antibody or Alexa Fluor-488 phalloidin (Invitrogen). Fluorescence images were captured via confocal laser scanning microscopy (C1 Confocal Imaging System; Nikon). Max-intensity z-projected images were obtained and analyzed using the Image J software.

Yamaguchi N., Mizutani T., Kawabata K, & Haga H. (2015). Leader cells regulate collective cell migration via Rac activation in the downstream signaling of integrin β1 and PI3K. Scientific Reports, 5, 7656.

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Quinone-Induced Oxidative Stress Assay

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Quinone-induced oxidative stress was determined using CellROX^® Green reagent (Life Technologies, Carlsbad, CA USA) in combination with confocal microscopy. For these studies (sub-confluent) N27 and clone 4 cells were grown on glass coverslips in 6-well plates. After treatment with quinones the medium was removed and cells washed three times with PBS. The CellROX^® Green fluorogenic probe (5µM in PBS) was added to cells for 30min at 37°C in the dark. Cells were washed three times with PBS then fixed in 3.7% (v/v) formaldehyde in PBS for 12min, and then permeabilized with 0.1% (v/v) Triton X-100 in PBS for 10min. After extensively washing with PBS cells were incubated in blocking buffer (RPMI-1640 containing 10% FBS) for 1h, and then incubated with anti-NQO1 antibody (1:100) overnight at 4°C followed by Alexa Fluor 594 conjugated goat anti-rabbit IgG (1:1000) containing DAPI (1µg/ml) for 30min. Coverslips were washed three times with TBST then rinsed in distilled water, inverted and mounted on glass slides using SuperMount (BioGenex, USA). Cells were viewed on a Nikon TE2000 microscope with a Nikon C1 confocal imaging system.

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