C2 confocal microscope system

Manufactured by Nikon

Sourced in Japan

The Nikon C2+ confocal microscope system is a high-performance imaging platform designed for advanced microscopy applications. It features a modular design, allowing for customization to meet the diverse needs of researchers and scientists. The C2+ provides optical sectioning capabilities, enabling the acquisition of clear, high-resolution images by reducing out-of-focus light. This system is capable of capturing detailed images of biological samples, making it a valuable tool for a wide range of research fields.

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Lab products found in correlation

Alexa fluor 488 conjugated donkey anti chicken, by Jackson ImmunoResearch (2 mentions) Fluoromount g, by Southern Biotech (2 mentions) Cy3 conjugated donkey anti rabbit, by Jackson ImmunoResearch (2 mentions) Nis elements imaging software, by Nikon (2 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (2 mentions) Superfrost plus slide, by Thermo Fisher Scientific (2 mentions) Nis elements software, by Nikon (2 mentions) Alexa fluor 488 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594 conjugated donkey anti mouse igg, by Thermo Fisher Scientific (2 mentions) Rabbit anti pax2, by Thermo Fisher Scientific (1 mentions) Gfp 1020, by Abcam (1 mentions) Rabbit anti c fos, by Abcam (1 mentions) Alexa fluor 555, by Thermo Fisher Scientific (1 mentions) Gfp 1020, by Merck Group (1 mentions) Cy5 conjugated donkey anti guinea pig, by Jackson ImmunoResearch (1 mentions) Ab153, by Merck Group (1 mentions) Goat anti mouse igg h l alexa fluor 647, by Abcam (1 mentions) Bovine serum albumin (bsa), by MP Biomedicals (1 mentions) Eos rebel t3, by Canon (1 mentions) R buffer a, by Electron Microscopy Sciences (1 mentions)

Spelling variants of this product

Confocal microscope (523 citations) C2 confocal microscope (379 citations) C2 confocal system (34 citations) C2 microscope (34 citations) C2 confocal (20 citations) C2+ system (17 citations) C2s system (5 citations)

31 protocols using c2 confocal microscope system

Immunofluorescence Staining of Mouse Brain Tissue

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Mice were deeply anesthetized with isoflurane and perfused intracardially with 4% paraformaldehyde in PBS⁶⁴. Tissues were dissected, post-fixed for 8 h, and cryoprotected in 20% sucrose overnight at 4 °C. Free-floating frozen sections were blocked in a 0.01 M PBS containing 2% donkey serum and 0.1% Triton X-100 followed by incubation with primary antibodies overnight at 4 °C and secondary antibodies for 2 h at room temperature. Sections were mounted with FluoromountG (Southern Biotech). The following primary antibodies were used: chicken anti-GFP (1:500, Aves Labs, GFP-1020), rabbit anti-c-Fos (1:4000, Abcam, ab190289), rabbit anti-lmx1b (1:500, Invitrogen, Cat.PA5-34471), rabbit anti-Pax2 (1:500, Invitrogen, Cat.71-6000) and rabbit anti-NKB (1:500, Novus, Cat.NB300-201). The following secondary antibodies were used: Alexa-Fluor 488 conjugated donkey anti-chicken (1:500, Jackson ImmunoResearch, 703-545-155), Cy3-conjugated donkey anti-rabbit (1:500, Jackson ImmunoResearch, 711-165-152) and 488-conjugated donkey anti-rabbit (1:500, Jackson ImmunoResearch, 711-545-152). Fluorescent Images were taken using a Nikon C2+ confocal microscope system (Nikon Instruments, Inc.).

Chen S., Gao X.F., Zhou Y., Liu B.L., Liu X.Y., Zhang Y., Barry D.M., Liu K., Jiao Y., Bardoni R., Yu W, & Chen Z.F. (2020). A spinal neural circuitry for converting touch to itch sensation. Nature Communications, 11, 5074.

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Phospho-JAK1 Immunocytochemistry in Mouse DRG Neurons

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Mouse DRG neurons were extracted and fixed in 4% paraformaldehyde (PFA) for 5 minutes on ice. The fixed cells were then stained overnight with rabbit anti-mouse/human phospho-JAK1 (Tyr1022, Tyr1023) primary antibody (59H4L5, Invitrogen) at 4°C. Secondary goat anti-rabbit antibody conjugated to Alexa Fluor 555 (Invitrogen) was used to stain for 1 hour at 4°C. VECTASHIELD mounting medium with DAPI was used for mounting and staining of nuclei (Vector Laboratories). All imaging and analysis was performed on a Nikon C2+ Confocal Microscope System with NIS-Elements imaging software (Nikon Instruments).

Oetjen L.K., Mack M.R., Feng J., Whelan T.M., Niu H., Guo C.J., Chen S., Trier A.M., Xu A.Z., Tripathi S.V., Luo J., Gao X., Yang L., Hamilton S.L., Wang P.L., Brestoff J.R., Council M.L., Brasington R., Schaffer A., Brombacher F., Hsieh C.S., Gereau RW I.V., Miller M.J., Chen Z.F., Hu H., Davidson S., Liu Q, & Kim B.S. (2017). Sensory Neurons Co-opt Classical Immune Signaling Pathways to Mediate Chronic Itch. Cell, 171(1), 217-228.e13.

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Immunofluorescence Labeling of Mouse Spinal Cord

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The procedures were described previously^{37 (link),62}. Deeply anesthetized mice (ketamine, 90 mg/kg and Xylazine, 10 mg/kg) were perfused transcardially with 0.01 M PBS (PH 7.4) and paraformaldehyde (PFA) (4% in PBS). Spinal cord and brain were removed and post-fixed in 4% PFA for 2–4 h. The tissues were then cryoprotected in 20% sucrose overnight at 4 °C. Free-floating frozen sections were incubated with 2% donkey serum and 0.3% Triton X-100 for 1 h at room temperature followed by incubation with primary antibodies overnight at 4 °C. The sections were then washed and incubated with secondary antibodies for 2 h at room temperature. The following primary antibodies were used: chicken anti-GFP (1:500, Aves Labs, GFP-1020), guinea-pig anti-NK1R (1:500, AB15810, EMD Millipore), rabbit anti-FG (1:5000, AB153, Millipore). The following secondary antibodies were used: Alexa-Fluor 488 conjugated donkey anti-chicken (1:1000, Jackson ImmunoResearch, 703–545–155), Cy3-conjugated donkey anti-rabbit (1:1000, Jackson ImmunoResearch, 711–165–152) and Cy5-conjugated donkey anti-guinea pig 1(:1000, Jackson ImmunoResearch, 703–175–148). Fluorescent Images were taken using a Nikon C2+ confocal microscope system (Nikon Instruments, Inc.).

Bardoni R., Shen K.F., Li H., Jeffry J., Barry D.M., Comitato A., Li Y.Q, & Chen Z.F. (2019). Pain Inhibits GRPR Neurons via GABAergic Signaling in the Spinal Cord. Scientific Reports, 9, 15804.

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Single-Molecule RNA Detection in Mouse Brain

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A high amplification system for the detection of single-molecule RNA fluorescence in situ hybridization, RNAscope® (Advanced Cell Diagnostics (ACD), Hayward, CA), was used for verification of iCre mRNA expression in the SCN and PVT of Grpr^iCre mice, Adcyap1r1 expression of Grpr^iCre mice and the co-localization of Adcyap1r1 and Grp in SCN. The tissue preparation procedure was the same as described above for the IHC experiments. Brain tissues were sectioned at 20 μm thickness in OCT using a cryostat microtome and mounted directly on SuperFrost Plus slides (Thermo Fisher Scientific, Waltham, MA). RNAscope experiment was performed according to our previous studies (Munanairi et al., 2018 (link)). Mouse-specific probes for Grpr (317871-C2, ACD) and iCre (423321-C3, ACD) Adcyap1r1 (409561-C1, ACD), and Grp (317861-C2, ACD) were purchased from Advanced Cell Diagnostics. Fluorescent Images were taken using a Nikon C2+ confocal microscope system (Nikon Instruments, Inc.) and analysis of images was performed using ImageJ software from NIH Image (version 1.34e). For each group of treatment, three sections were averaged in each mouse, and 3~6 mice were used for statistical analysis. Cell numbers in the entire SCN were counted.

Gao F., Ma J., Yu Y.Q., Gao X.F., Bai Y., Sun Y., Liu J., Liu X., Barry D.M., Wilhelm S., Piccinni-Ash T., Wang N., Liu D., Ross R.A., Hao Y., Huang X., Jia J.J., Yang Q., Zheng H., van Nispen J., Chen J., Li H., Zhang J., Li Y.Q, & Chen Z.F. (2022). A non-canonical retina-ipRGCs-SCN-PVT visual pathway for mediating contagious itch behavior. Cell reports, 41(1), 111444.

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Immunofluorescent Labeling of Rat Hippocampus

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The hippocampal sections from rats were fixed in 4% paraformaldehyde for 10 min, permeabilised with 0.4% Triton X-100 for 15 min, and incubated in 5% bovine serum albumin (MP Biomedicals, CA) for 30 min at 37 °C. The sections were then incubated overnight at 4 °C with a mixture of rabbit polyclonal anti-ASIC2a antibody (1:100, Abcam) and mouse monoclonal anti-TFCP2 antibody (1:100, BD Biosciences). They were then incubated in a darkroom for 90 min at 37 °C with goat anti-rabbit IgG H&L (Alexa Fluor® 488, Cat. No. ab150077, Abcam) and goat anti-mouse IgG H&L (Alexa Fluor®647, Cat. No. ab150115, Abcam) antibodies and mounted in 1:1 glycerol/phosphate-buffered saline (PBS). The sections from AAV-injected rats were obtained 3 weeks after AAV infusion. Nuclei were stained with DAPI (300 nM) for 15 min.
PC12 cells were fixed in 4% paraformaldehyde for 20 min, permeabilised with 0.4% Triton X-100 for 15 min, and incubated with 5% goat serum for 30 min at 37 °C. The cells were strained as before. Nuclei were stained with DAPI (300 nM) for 15 min and then rinsed in PBS. Confocal images were acquired with a Nikon C2⁺ confocal microscope system (Nikon, Japan).

Zhang H., Gao G., Zhang Y., Sun Y., Li H., Dong S., Ma W., Liu B., Wang W., Wu H, & Zhang H. (2017). Glucose Deficiency Elevates Acid-Sensing Ion Channel 2a Expression and Increases Seizure Susceptibility in Temporal Lobe Epilepsy. Scientific Reports, 7, 5870.

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Planarian Microscopy Imaging Protocol

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Planarians and processed samples analyzed by bright and dark field microscopy, as well as those analyzed by low-magnification fluorescence microscopy, were photographed using an Axio Zoom V16 stereomicroscope (Zeiss, Oberkochen, Germany) equipped with an EOS Rebel T3 digital camera (Canon, Tokyo, Japan). High-magnification immunofluorescence analyses were carried out by confocal microscopy under a 10×, 20×, or oil-immersion 60× objective in a Nikon C2+ Confocal Microscope System. Z-stacks were generated from image sectioning of samples every 2– 3 microns and assembled using the NIS Elements Imaging Software (Nikon Corporation, Tokyo, Japan) to produce maximum projection and three-dimensional images. Brightness and contrast were adjusted for some images without producing changes that would alter interpretation of data.

Almazan E.M., Ryan J.F, & Rouhana L. (2021). Regeneration of Planarian Auricles and Reestablishment of Chemotactic Ability. Frontiers in Cell and Developmental Biology, 9, 777951.

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Immunohistochemistry Protocol for Phospho-Protein Analysis

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Samples were fixed in 10% formalin and embedded in paraffin. After deparaffinization, slides were washed with 9.83% NaCl for 3 min followed by a PBS wash and a wash in distilled water for 5 min. Heat induced antigen retrieval was performed with pressure cooker (2100 Retriever, PickCell Laboratories B.V.) and R-Buffer A (pH6.0, Electron Microscopy Sciences). Sections were incubated with 2% H₂O₂ in Methanol for 15 minutes for endogenous peroxidase quenching and washed and blocked for non-specific binding in 1% BSA in PBS-Triton 0.3% v/v (PBST) for 1 hour. Subsequently, sections were sequentially incubated with primary antibody at 1:100 dilution for 1 hour, secondary peroxidase goat anti-rabbit IgG antibody (Vector) at 1:250 dilution for 1 hour and Tyramide Signal Amplification (TSA) Fluorescein System (Perkin Elmer, Cat.: NEL701A) according to kit manual. Slides were mounted with Vectashield mounting medium with DAPI (Vector), photographed with Nikon C2 Confocal Microscope system and subsequently stained with Hematoxylin and Eosin. p-AKT (Thr308) p-ERK (Thr202/Tyr204), p-S6 (Ser235/236), and CC3 (Asp 175) were obtained from Cell Signaling, Inc. Ki67 staining was performed as previously described (19 (link)). Quantification of Ki67 staining was performed by scoring the nuclei of cells from each lesion type found in a minimum of ten high-powered fields.

Alagesan B., Contino G., Guimaraes A.R., Corcoran R.B., Deshpande V., Wojtkiewicz G.R., Hezel A.F., Wong K.K., Loda M., Weissleder R., Benes C.H., Engelman J, & Bardeesy N. (2014). Combined MEK and PI3K inhibition in a mouse model of pancreatic cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(2), 396-404.

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Multimodal Imaging Techniques for Comprehensive Sample Analysis

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DAB immunohistochemical images were acquired using a Leica DM5500 B upright microscope with 5× objective and Leica DFC425 C digital camera with Leica Application Suite (version 4.4.0) acquisition software or a Leica SCN400 slide scanner at ×40 magnification. Toluidine blue staining images were acquired using a Leica SCN400 slide scanner at ×40 magnification. Fluorescent confocal images were acquired using a Nikon C2 confocal microscope system (Nikon, Melville, NY) with NIS-Elements software (Nikon, Melville, NY). Confocal images for 3D reconstruction were captured with 0.3-μm step size using the following objectives: Plan Fluor 40×/1.30 numerical aperture (NA) (oil immersion) and Apo 60×/1.40 NA (oil immersion). An advanced noise removal algorithm was applied evenly across experimental groups. For electron microscopy, grids were examined on a Tecnai G2 SpiritBT transmission electron microscope (FEI Company, Hillsboro, OR) operated at 60 kV.

Evonuk K.S., Doyle R.E., Moseley C.E., Thornell I.M., Adler K., Bingaman A.M., Bevensee M.O., Weaver C.T., Min B, & DeSilva T.M. (2020). Reduction of AMPA receptor activity on mature oligodendrocytes attenuates loss of myelinated axons in autoimmune neuroinflammation. Science Advances, 6(2), eaax5936.

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Immunohistochemical Analysis of c-Fos and NPY

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Immunohistochemistry (IHC) was performed as previously described.^{23 (link)} After c-Fos induction, mice were anesthetized (pentobarbital, 50 mg/kg, i.p.) and perfused intracardially with 0.01 M PBS pH 7.4 followed by 4% paraformaldehyde (PFA) in PBS. Spinal cord tissues were dissected, post-fixed for 6-8 h, and cryoprotected in 20% sucrose in PBS overnight at 4°C. Subsequently, tissues were embedded in optimum cutting temperature medium (Tissue-Plus O.C.T. compound, Fisher Healthcare) and were cut at 30 µm by a Leica CM1950 cryostat. Free-floating frozen sections were blocked for 1 h in a 0.01 M PBS solution containing 2% donkey serum and 0.1% Triton X-100 followed by incubation with primary antibody (rabbit anti-c-Fos, 1:4000, Abcam, ab190289; goat anti-NPY, 1:1000, Novus, NBP1-46535) overnight at 4°C, washed three times with PBS, secondary antibody (488-conjugated donkey anti-rabbit, 1:500, Jackson ImmunoResearch, 711-545-152; 594-conjugated donkey anti-goat, 1:500, Jackson ImmunoResearch 705-585-147) for 2 h at room temperature and washed again for three times. Sections were mounted on slides and ∼200 μL FluoromountG (Southern Biotech) was placed on the slide with a coverslip. Fluorescent Images were taken using a Nikon C2 + confocal microscope system (Nikon Instruments, Inc.).

Chen S., Chen J., Tang D., Yin W., Xu S., Gao P., Jiao Y, & Yu W. (2024). Mechanical and chemical itch regulated by neuropeptide Y-Y1 signaling. Molecular Pain, 20, 17448069241242982.

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Multimodal Imaging of Myelination in EAE

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Images for DAB immunohistochemistry of MBP in the spinal cord and optic nerve were acquired using a Leica SCN400 slide scanner at 40 × magnification. Fluorescent images were acquired using a Nikon C2 confocal microscope system (Nikon, Melville, NY) with NIS-Elements software (Nikon, Melville, NY) at 40x (for sectioned retinas, optic nerve, and spinal cord white matter), 20x (for thalamic regions-dorsal lateral geniculate nucleus (dLGN) and ventral posterolateral nucleus (VPL), and spinal cord gray matter), or 10x (for NeuN in the thalamus and spinal cord gray matter) magnification. For retinal whole mounts, confocal z-stacks of Brn3a⁺ cells were captured in the retinal ganglion cell layer at 20 × magnification with 2.50 µm thickness and 0.5 µm step size. For qualitative images of EAE lesions in the spinal cord and optic nerve, 10 µm-thick confocal z stacks were captured at 20 × and 40x, and 3D reconstructions were generated for representative images. For electron microscopy, grids were examined on a Tecnai G2 SpiritBT transmission electron microscope (FEI Company, Hillsboro, OR) operated at 60 kV. NIS-Elements software (Nikon, Melville, NY) and Adobe Photoshop (for contrast, brightness, and color adjustments) were used to create figures and process images.

Mey G.M., Evonuk K.S., Chappell M.K., Wolfe L.M., Singh R., Batoki J.C., Yu M., Peachey N.S., Anand-Apte B., Bermel R., Ontaneda D., Nakamura K., Mahajan K.R, & DeSilva T.M. (2022). Visual imaging as a predictor of neurodegeneration in experimental autoimmune demyelination and multiple sclerosis. Acta Neuropathologica Communications, 10, 87.

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