Spinning disk confocal microscope

Manufactured by Leica

Sourced in Germany

The Spinning disk confocal microscope is a specialized optical microscope that uses a spinning disk with pinholes to rapidly scan a sample and produce high-resolution, three-dimensional images. It functions by illuminating the sample with a laser and rapidly acquiring images through the pinholes in the spinning disk, allowing for fast and efficient capture of data.

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Lab products found in correlation

Glass coverslip, by Avantor (2 mentions) Fluoromount g mounting solution, by Southern Biotech (2 mentions) Las af program software, by Leica (2 mentions) Duolink in situ red kit mouse rabbit, by Merck Group (2 mentions) Biorevo bz 9000 digital microscope, by Keyence (1 mentions) Cytokeratin 5, by Fortrea (1 mentions) Sp5 confocal microscope, by Leica (1 mentions) Hoechst, by AnaSpec (1 mentions) Anti e cad, by Cell Signaling Technology (1 mentions) Anti k8, by Developmental Studies Hybridoma Bank (1 mentions) Anti k14, by Fortrea (1 mentions) Pluronic f 127, by Thermo Fisher Scientific (1 mentions) Fluo 4, by Thermo Fisher Scientific (1 mentions) R skf38393, by Merck Group (1 mentions) Ab144p, by Merck Group (1 mentions) Aqi x combo cwf program, by Media Cybernetics (1 mentions) Lab tek chambered cover glasses, by Thermo Fisher Scientific (1 mentions) Human fibronectin, by Merck Group (1 mentions) Lsm 510 meta confocal module, by Zeiss (1 mentions) Eclipse ti e tirf microscope, by Nikon (1 mentions)

Close spelling variants of this product

Confocal microscope (1146 citations) Spinning disk microscope (2 citations) Spinning-disk confocal microscope system (2 citations) Confocal spinning disk inverted microscope (2 citations)

23 protocols using spinning disk confocal microscope

Multimodal Microscopic Analysis of Cellular Markers

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IHC was performed as previously described with: Hoechst (AnaSpec, #AS-83218), anti-E-cad (Cell Signaling, #3195 S or BD biosciences, 610182), anti-VANGL2 (SCBT, #46561), anti-VANGL1 (R&D, #HPA02235), Ki67 (Novus, NB500-170), Cleaved Caspase 3 and pERM (Cell Signaling #9661, #3726), Phalloidin (Invitrogen, #A12381), aSMA (DAKO, #M0851), anti-K8 (Developmental Studies Hybridoma Bank, Troma-1), Cytokeratin 5 (Covance, #PRB-160P), and anti-K14 (Covance, #PRB-155P). Images were collected on a Leica SP5 confocal microscope. Brightfield imaging was performed on a Biorevo BZ-9000 Digital Microscope (Keyence) and confocal microscopy performed on a Nikon C2 Confocal, Leica SP5 confocal and Solamere Spinning Disk Confocal Microscope. 3-dimensional reconstructions were performed using Fiji or NIS-elements.

Smith P., Godde N., Rubio S., Tekeste M., Vladar E.K., Axelrod J.D., Henderson D.J., Milgrom-Hoffman M., Humbert P.O, & Hinck L. (2019). VANGL2 regulates luminal epithelial organization and cell turnover in the mammary gland. Scientific Reports, 9, 7079.

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Calcium Imaging of Cultured Neurons

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On the seventh day, cultured neurons were loaded for 35 min at RT with a calcium probe Fluo-4 and pluronic F 127 (v/v; 50%; Invitrogen) diluted to 1/500 in a glutamate-free recording medium (129 mM NaCl, 4 mM KCl, 1 mM MgCl₂, 2 mM CaCl₂, 10 mM Glucose, 10 mM HEPES). Neurons were washed three times in this recording medium before imaging. Acquisitions were performed at 36.2 °C every second, prior to and during treatments with the indicated doses of (R)-( + )-SKF38393 (Sigma Aldrich) and/or L-glutamic acid (Calbiochem), with a spinning disk confocal microscope (Leica) using a 40X oil immersion objective. Fluorescence intensity was analyzed in the soma of all neurons present in each field by using the Fiji software. Viability of the neurons was tested after stimulation with a high dose of glutamate (10 μM), which yields high and sustained calcium responses, at the end of image acquisition. Data are presented as means ± s.e.m. of ΔF/F, ([Ft-F0]/F0 where Ft and F0 are the fluorescence intensities after and before stimulation, respectively. Representative pictures of the ΔF/F signal at the pic of the responses were prepared with MatLab (Version 9.4 R2018a). Calcium imaging experiments were independently performed on 5 littermates. 5 to 23 µ-wells were analyzed for each pharmacological treatment.

Zayed A., Baranowski C., Compagnion A.C., Vernochet C., Karaki S., Cuttoli R.D., Saint-Jour E., Bhattacharya S., Marti F., Vanhoutte P., Yaniv M., Faure P., Barik J., Amar L., Tronche F, & Parnaudeau S. (2022). SWI/SNF chromatin remodeler complex within the reward pathway is required for behavioral adaptations to stress. Nature Communications, 13, 1807.

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Immunofluorescence for Cholinergic Neurons

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Immunofluorescence staining and confocal microscopy was used to determine mouse anti-Chat (AB144P, Chemicon, Tumecula CA USA). Images were analyzed using a spinning disk confocal microscope (Leica, Buffalo Grove IL USA). Deconvolution and 3-dimensional construction of the confocal image was performed by AQI-X-COMBO-CWF program (Media cybernetics Inc., Rockville, MD, USA). Control experiments were performed in the absence of primary antibody or in the presence of blocking peptide.

Kim K.Y., Kim Y.R., Choi K.W., Lee M., Lee S., Im W., Shin J.Y., Kim J.Y., Hong Y.H., Kim M., Kim J.I, & Sung J.J. (2020). Downregulated miR-18b-5p triggers apoptosis by inhibition of calcium signaling and neuronal cell differentiation in transgenic SOD1 (G93A) mice and SOD1 (G17S and G86S) ALS patients. Translational Neurodegeneration, 9, 23.

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Live-Cell Imaging of Neutrophil Chemotaxis

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For live-cell imaging, HL-60 neutrophils were collected, washed and resuspended with mHBSS containing 0.2% BSA, and then seeded on Lab-Tek chambered cover glasses (NUNC) coated with 50 μg/ml human fibronectin (Sigma-aldrich). Time-lapse microscopy was performed on a Leica Spinning Disk Confocal microscope. Chemoattractant fMLP (Sigma) was given to cells either globally or delivered by a Femtotips micropipette (Eppendorf). Total Internal Reflection Fluorescence (TIRF) microscopy was performed using a Nikon Eclipse Ti-E TIRF microscope. Fluorescence recovery after photobleaching (FRAP) was performed on Zeiss Axiovert 200 Laser scanning microscope with LSM510-Meta confocal module. For immuno uorescent staining, cells were xed, and then stained with Anti-PI 3-Kinase, p110γ (clone 17D7.2, Merck Millipore) in PBS containing 1% BSA as indicated in the Extended Experimental Procedures. Images were analyzed with ImageJ (NIH) as described in the Extended Experimental Procedures.

Wang M., Artemenko Y., Cai W., Iglesias P.A, & Devreotes P.N. (2014). The Directional Response of Chemotactic Cells Depends on a Balance between Cytoskeletal Architecture and the External Gradient. Cell reports, 9(3), 1110-1121.

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Immunohistochemical Analysis of Muscle Fibers

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Immediately after sacrificing the animal, the skin of the leg was surgically removed and fascia from the TA muscle was taken off. The whole leg was severed and incubated in fresh 4% PFA for 1 h at room temperature. The leg was washed 3 × 5 min in PBS and stored in PBS containing 0.02% sodium azide at 4°C. Fixed muscles were teased into single fibers using fine forceps. Non-specific staining was blocked with 2% bovine serum albumin (BSA) and 2% goat serum in PBS supplemented with 0.5% Triton-X100. The fibers were incubated with primary antibodies overnight in the same buffer. Fibers were washed 3 × 5 min in PBS followed by incubation with secondary antibodies and fluorescently labeled bungarotoxin in 2% BSA and 2% goat serum in PBS containing 0.1% Triton-X100 for 1 h at room temperature. Cells were washed 3 × 5 min in PBS and mounted in fluoromount with DAPI, covered with coverslips, and sealed with nail polish. Image acquisition was performed at the Confocal Microscopy Facility at the Nencki Institute, using the Zeiss spinning disk confocal microscope or the Leica TCS SP8 scanning confocal microscope. Images were analyzed using ImageJ/Fiji software.

Bernadzki K.M., Daszczuk P., Rojek K.O., Pęziński M., Gawor M., Pradhan B.S., de Cicco T., Bijata M., Bijata K., Włodarczyk J., Prószyński T.J, & Niewiadomski P. (2020). Arhgef5 Binds α-Dystrobrevin 1 and Regulates Neuromuscular Junction Integrity. Frontiers in Molecular Neuroscience, 13, 104.

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Visualizing Migratory Cells in PRP Fibers

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Migratory cells were visualized 1 to 2.5 mm above the bottom of the plate in
which the PRP fibers were formed by phase-contrast microscopy using a spinning
disk confocal microscope (Leica). Phase-contrast images were stitched together
using Volocity 3D Image Analysis software (Quorum Technologies).

Wu M.J., Sermer C., Kandel R.A, & Theodoropoulos J.S. (2022). Characterization of Migratory Cells From Bioengineered Bovine Cartilage in a 3D Co-culture Model. The American Journal of Sports Medicine, 50(11), 3090-3101.

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Immunostaining Protocol for Confocal Microscopy

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In brief, the tissues/sections were permeabilized in ice-cold 70% ethanol (Sigma-Aldrich) for 10min at −20°C and washed with 1% Triton X-100 (Sigma-Aldrich) in TBS (TBS-T) 3 times for further immunostaining. The specimens were first incubated in blocking buffer (1% FBS, 3% BSA, 0.5% Triton-X100, 0.01% sodium deoxycholate in TBS) for 30min at room temperature and subsequently incubated in blocking buffer containing antibodies for overnight at 4°C. Next day, specimens were washed 3 times with TBS-T (10 min/wash), incubated in secondary antibody at room temperature for 1hr. After they were washed three times with TBS-T in the dark (10 min/wash), the specimens were mounted on glass slides with anti-fading mounting medium (10% 0.1M Tris buffer (pH 9.0), 0.2mg/mL p-phenylenediamine hydrochloride (Sigma-Aldrich) and 4mM sodium azide (Sigma-Aldrich) in glycerol). Confocal microscopy was performed with Nikon ECLIPSE 80i fluorescence microscope, Leica spinning disk confocal microscope or a Leica SP8 confocal microscope. ImageJ (NIH) was used for the data analysis

Lee H.W., Xu Y., He L., Choi W., David G., Jin S.W, & Simons M. (2021). The Role of Venous Endothelial Cells in Developmental and Pathologic Angiogenesis. Circulation, 144(16), 1308-1322.

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Confocal Imaging of Cells in Collagen

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Images are acquired on a spinning disk confocal microscope (Leica, Solms, Germany) with an electron multiplying CCD camera (Hamamatsu Photonics, Hamamatsu, Japan). A 63x 1.4NA oil immersion objective and dual lasers for exciting GFP and mCherry are used. Micro-Manager Software is used for the image acquisition. Because the image capture is 100 seconds per cell, no large focal drifts are observed. The change of collagen concentration alone did not appear to affect the image quality. Representative images of the cell and mitochondria in 1–4 mg/mL collagen gels can be found in Fig. S2.

Kim J.E., Reynolds D.S., Zaman M.H, & Mak M. (2018). Characterization of the Mechanical Properties of Cancer Cells in 3D Matrices in Response to Collagen Concentration and Cytoskeletal Inhibitors. Integrative biology : quantitative biosciences from nano to macro, 10(4), 232-241.

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Immunofluorescence Imaging of Serine Metabolism

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Cells were cultured in conditional media containing 0.8 or 0 mM serine with an addition of 10% FBS on glass coverslips (VWR) in six-well dishes for 24 hr. Next, cells were fixed with 4% paraformaldehyde for 15 min at room temperature. After one wash in 1× PBS, cells were permeabilized in 0.15% Triton X-100 for 15 min at room temperature. Cells were then blocked in 10% BSA for 20 min at room temperature, followed by incubation in primary antibody solution (1:500 dilution in 10% BSA) for overnight at 4°C. After three washes using 1× PBS, cells were incubated in secondary antibody solution in 10% BSA and followed by another three times of wash. Cells were then mounted onto glass slides using Fluoromount-G mounting solution (Southern Biotech) for imaging acquisition on a spinning-disk confocal microscope (Leica) using a × 100/1.4NA oil or a ×40/1.25NA oil (Leica Plan Apochromat) objective. Images were acquired and processed using the Leica LAS AF program software.

Gao X., Lee K., Reid M.A., Sanderson S.M., Qiu C., Li S., Liu J, & Locasale J.W. (2018). Serine Availability Influences Mitochondrial Dynamics and Function through Lipid Metabolism. Cell reports, 22(13), 3507-3520.

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Aortic CD11c+ Immune Cell Imaging

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Ldlr^−/− mice and C57BL/6 naïve mice (as controls) were injected intravenously with 150 μL of PS (15 mg/mL) as previously described. After 24 h, the heart and aorta were perfused with 4% paraformaldehyde (PFA)/5% sucrose in PBS solution for 10 min. The aorta was harvested and fixed in 4% PFA/5% sucrose PBS solution 12 h at 4 °C. The isolated aortic arch-derived arteries were immersed in 15% sucrose solution for 12 h and then 30% sucrose solution for 24 h. The resulting specimens were embedded in Tissue-Tek OCT and frozen at −80 °C. Tissue blocks were sectioned at 5 μm thickness and stained with 4′,6-diamidino-2-phenylindole (DAPI) and Alex-488-anti-CD11c antibody. Images were taken on a spinning disk confocal microscope (Leica). Z-stacks were performed at the inner aortic tissue at 63× magnification using MetaMorph software.

Yi S., Allen S.D., Liu Y.G., Ouyang B.Z., Li X., Augsornworawat P., Thorp E.B, & Scott E.A. (2016). Tailoring Nanostructure Morphology for Enhanced Targeting of Dendritic Cells in Atherosclerosis. ACS nano, 10(12), 11290-11303.

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