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Spinning diskmicroscope

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The Spinning diskmicroscope is a type of microscope that uses a spinning disk to illuminate and image samples. It is designed to capture high-quality, high-resolution images of specimens by rapidly scanning a laser beam across the sample. The spinning disk creates an array of illumination points, allowing for efficient light collection and fast image acquisition.

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Lab products found in correlation

Micropoint, by Oxford Instruments (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Em ccd camera, by Hamamatsu Photonics (1 mentions) Volocity 6, by PerkinElmer (1 mentions) Nanog, by Cell Signaling Technology (1 mentions) Tnnt2, by Thermo Fisher Scientific (1 mentions) Mef2c, by Cell Signaling Technology (1 mentions) Actn2, by Merck Group (1 mentions) Dental sg resin, by Formlabs (1 mentions)

7 protocols using spinning diskmicroscope

1

Laser Wound Imaging Protocol

Cited 1 time
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Laser wounding was performed using an ablation laser (MicroPoint; Andor
Technology) as previously described70 (link) and imaged using a Perkin Elmer spinning disk
microscope.
Yolland L., Burki M., Marcotti S., Luchici A., Kenny F.N., Davis J.R., Serna-Morales E., Müller J., Sixt M., Davidson A., Wood W., Schumacher L.J., Endres R.G., Miodownik M, & Stramer B.M. (2019). Persistent and polarised global actin flow is essential for directionality during cell migration. Nature cell biology, 21(11), 1370-1381.
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2

Optical Manipulation of Cell Junctions

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Optical manipulation of the cell junctions in individual embryos was done using a spinning-disk microscope (Perkin-Elmer), coupled with a home-built two-point manipulation laser system. The laser trap system was described in (30 (link)). A 100× water immersion lens (CFI Plan Apochromat VC 100× oil, NA 1.40, WD 0.13) was used for imaging and optical manipulation in the imaging plane. To achieve two-point manipulation, we split an IR laser beam (1,070-nm wavelength) into two beams by fast commutation of two galvanometric mirrors. The switching time of the galvanometric mirrors is 100 µs, representing 2.5% of the time spent on each trap (4 ms). The relationship between the (X and Y) positions of the two resulting traps and the galvanometric command voltages is calibrated by trapping colloidal beads in water prior to manipulation in the embryo (30 (link)).
Prior to a junction manipulation, laser traps are stably positioned for 5 s on a point located along the target junction. The trap is then moved to a defined distance and maintained (Fig. 1A). If not otherwise stated, the trap displacements were 1.5 µm within 3 s, the laser power per trap was fixed at 200 mW, and the acquisition was performed over 3 min.
Nishizawa K., Lin S.Z., Chardès C., Rupprecht J.F, & Lenne P.F. (2023). Two-point optical manipulation reveals mechanosensitive remodeling of cell–cell contacts in vivo. Proceedings of the National Academy of Sciences of the United States of America, 120(13), e2212389120.
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3

Laser Wound Imaging Protocol

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Laser wounding was performed using an ablation laser (MicroPoint; Andor
Technology) as previously described70 (link) and imaged using a Perkin Elmer spinning disk
microscope.
Yolland L., Burki M., Marcotti S., Luchici A., Kenny F.N., Davis J.R., Serna-Morales E., Müller J., Sixt M., Davidson A., Wood W., Schumacher L.J., Endres R.G., Miodownik M, & Stramer B.M. (2019). Persistent and polarised global actin flow is essential for directionality during cell migration. Nature cell biology, 21(11), 1370-1381.
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4

GFP-Fusion Protein Localization in HEK 293T Cells

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The full length, truncated and mutated SPNS2 genes with N-terminal GFP fusion were cloned into pcDNA5/TO plasmid. The plasmid was transfected into HEK 293T cells for 24 h via lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol. Live cells were imaged with a Perkin Elmer Spinning Disk Microscope. 100×Olympus PlanApo Objective with Numerical Aperture of 1.4 was used. GFP was excited by the 488 nm laser and the fluorescence images were taken using EM-CCD camera (Hamamatsu). Images were analysed using Volocity 6.3.1 software (Perkin Elmer).
Tang H., Li H., Prakaash D., Pedebos C., Qiu X., Sauer D.B., Khalid S., Duerr K, & Robinson C.V. (2023). The solute carrier SPNS2 recruits PI(4,5)P2 to synergistically regulate transport of sphingosine-1-phosphate. Molecular Cell, 83(15), 2739-2752.e5.
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5

Immunofluorescence Staining of Stem Cells

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Cells were fixed with 4% paraformaldehyde (PFA) for 15 min, blocked for 1 h in blocking buffer (5% FBS, 0.25% Triton X‐100 in PBS), and stained for OCT4 (SantaCruz, #5279), NANOG (Cell Signaling, #3580S), TNNT2 (ThermoFisher Scientific, #MA5‐12960), ACTN2 (Sigma, #A7811), MEF2C (Cell Signaling, #sc‐13266), GFP (Abcam, #ab6556) in staining buffer (1% BSA, 0.25% Triton X‐100 in PBS). Nuclei were stained with Hoechst for 15 min. Images were made using a spinning disk microscope (PerkinElmer).
Dierickx P., Vermunt M.W., Muraro M.J., Creyghton M.P., Doevendans P.A., van Oudenaarden A., Geijsen N, & Van Laake L.W. (2017). Circadian networks in human embryonic stem cell‐derived cardiomyocytes. EMBO Reports, 18(7), 1199-1212.
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6

Customized Cell Compression Device

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A custom-made cell compression device has been invented based on movement of thin membrane attached with a piston, which is precisely controlled by air pressure regulator. The cell compression device was designed using Solidworks and device components were 3D-printed using Dental SG resin (Formlabs) for its biocompatibility. All the components were printed and then washed with IPA for 20 min, followed by post processing in ultraviolet chamber as suggested by Formlabs. A 20 mm diameter coverslip was stick on the top center of the cell compression device. Silicon membrane was sticked with a piston and then clamped to the bottom of the cell compression device by clamping tools. The assembled cell compression device was then connected to the air pressure regulator. Cells were plated on glass-bottom petridish and maintained in cell incubator. Before the experiment, a cell compression device was capped and locked on the cell culture dish. Images were acquired using ×40 oil lens (NA = 1.3) in PerkinElmer spinning disk microscope.
Kidiyoor G.R., Li Q., Bastianello G., Bruhn C., Giovannetti I., Mohamood A., Beznoussenko G.V., Mironov A., Raab M., Piel M., Restuccia U., Matafora V., Bachi A., Barozzi S., Parazzoli D., Frittoli E., Palamidessi A., Panciera T., Piccolo S., Scita G., Maiuri P., Havas K.M., Zhou Z.W., Kumar A., Bartek J., Wang Z.Q, & Foiani M. (2020). ATR is essential for preservation of cell mechanics and nuclear integrity during interstitial migration. Nature Communications, 11, 4828.
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7

HBV Infection Kinetics in Hepatocytes

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A total of 1 × 107 genome equivalent copies of HBV or HBV antigen-deficient virus supplemented with 4% polyethylene glycol 8000 (PEG 8000) were incubated with 2 × 104 HepG2-NTCP cells or PHH. The cells were maintained for 7 days subsequently in PMM supplemented with 2% FBS and changed to fresh medium every 2 days. For single-cell sequencing, the cells were collected after trypsin digestion. For antigen staining, the intracellular horseradish peroxidase was removed with phosphate-buffered saline (PBS) supplemented with 3% H2O2 and 40% CH3OH after the cells were fixed with 3.7% paraformaldehyde (PFA) and then stained for core with 1C10 Ab and for S with 56A1 Ab, followed by the phycoerythrin (PE) tyramide signal amplification (TSA) fluorescence system. The cell images were captured with a Nikon structured illumination microscope (SIM) or PerkinElmer spinning-disk microscope. Secreted viral antigen HBsAg and HBeAg were detected by ELISA.
Peng B., Jing Z., Zhou Z., Sun Y., Guo G., Tan Z., Diao Y., Yao Q., Ping Y., Li X., Ren T., Li B, & Li W. (2022). Nonproductive Hepatitis B Virus Covalently Closed Circular DNA Generates HBx-Related Transcripts from the HBx/Enhancer I Region and Acquires Reactivation by Superinfection in Single Cells. Journal of Virology, 97(1), e01717-22.
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