Ixon du888

Manufactured by Oxford Instruments

Sourced in United Kingdom, Ireland

The IXon DU888 is an electron-multiplying CCD (EMCCD) camera designed for low-light scientific imaging applications. It features a back-illuminated sensor with high quantum efficiency and low readout noise, enabled by its on-chip electron multiplication technology. The camera provides high-speed, high-resolution imaging capabilities.

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Lab products found in correlation

670 nm diode laser, by B&W Tek (2 mentions) Ckx41, by Olympus (1 mentions) X cite 120, by Olympus (1 mentions) Metafluor software control, by Molecular Devices (1 mentions) Diaphot 200 microscope, by Nikon (1 mentions) Fv1000 confocal laser scanning microscope, by Olympus (1 mentions) Csu10, by Yokogawa (1 mentions) Las x, by Leica (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Tcs sp8 confocal microscope, by Leica (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Perfect focus system, by Nikon (1 mentions) Eclipse ti2, by Oxford Instruments (1 mentions) Cfi plan apo lambda, by Nikon (1 mentions) Nis elements ar software, by Nikon (1 mentions) Deltavision software, by GE Healthcare (1 mentions) Deltavision omx, by GE Healthcare (1 mentions) Eclipse ti, by Nikon (1 mentions) Slowfade gold mounting media, by Thermo Fisher Scientific (1 mentions) Vectashield mounting media, by Vector Laboratories (1 mentions)

Spelling variants of this product

IXon° Du888 EMCCD camera (4 citations) IXon DU888 Ultra EMCCD camera (3 citations) IXon Ultra DU888 EMCCD (2 citations)

11 protocols using ixon du888

Netrin-1 Binding Substrate Analysis

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Netrin-1 attached on the substrate was analyzed as described (Moore et al., 2012 (link)) with modifications. Glass coverslips coated with polylysine or coated sequentially with polylysine and L1-CAM-Fc were incubated with culture medium and netrin-1 containing a 10-His tag at the C-terminus, in the absence or presence of 2 μg/ml heparin. The glasses were washed with PBS, and blocked for 1 hr at room temperature with 0.1% BSA in PBS. They were then labeled with anti-His antibody and secondary anti-mouse antibody conjugated with Alexa Fluor 488. Fluorescence images were acquired using a fluorescence microscope (IX81; Olympus) equipped with an EM-CCD camera (Ixon DU888; Andor), using a plan-Fluar 20 × 0.45 NA objective (Olympus), and quantified using ImageJ (Fiji version).

Baba K., Yoshida W., Toriyama M., Shimada T., Manning C.F., Saito M., Kohno K., Trimmer J.S., Watanabe R, & Inagaki N. (2018). Gradient-reading and mechano-effector machinery for netrin-1-induced axon guidance. eLife, 7, e34593.

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Fluorescent Imaging of Giant Vesicles

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Fluorescent live cell imaging and photography was performed using an Olympus CKX41 inverted light microscope with an X-cite 120 fluorescence camera and illumination system. Images were captured for 13 fields of views and the number of cells per field of view (x400 magnification) was counted, as determined by nuclear DAPI staining. Using FM® 1-43FX fluorescence, the number of giant vesicles (GV) (greater than 1µm in diameter) was counted in the same fields. Imaging was also performed using an Olympus IX81 with spinning disk confocal microscope and images taken using an Andor iXon DU888 camera.

Wright P.K., Jones S.B., Ardern N., Ward R., Clarke R.B., Sotgia F., Lisanti M.P., Landberg G, & Lamb R. (2014). 17β-estradiol regulates giant vesicle formation via estrogen receptor-alpha in human breast cancer cells. Oncotarget, 5(10), 3055-3065.

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Fluorescent Imaging of Cellular Structures

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Fluorescent images of the RINm-5F cells were acquired with a Nipkow spinning disk confocal system (Yokogawa CSU-10; Yokogawa Electric Corporation, Tokyo, Japan) attached to a Diaphot 200 microscope (Nikon, Kanagawa, Japan) and equipped with a 60×1.40/NA oil immersion objective (Nikon). Fluorescence was excited at 488 nm with argon ion laser (Melles-Griot, Didam, The Netherlands) and detected at 520/35 nm using cooled EM-CCD camera (iXon+ DU888; Andor, Belfast, Northern Ireland, UK) under MetaFluor software control (Molecular Devices Corp., Downingtown, PA).
Images of the human caudate nucleus were acquired by an Olympus FV1000 confocal laser scanning microscope (Olympus) equipped with a 20×/0.95W XLUMPlanFI objective. Emission spectra for each dye were limited as follows: Hoechst (361–497 nm) and Cy5 (630–647 nm). For co-localization analysis of PDYN and Hoechst z-stacks of 24 images were acquired with a depth interval of 0.5 µm. The open source platform Image J (NIH, MD) was used for 3D projections and orthogonal co-localization analysis.

Kononenko O., Bazov I., Watanabe H., Gerashchenko G., Dyachok O., Verbeek D.S., Alkass K., Druid H., Andersson M., Mulder J., Svenningsen Å.F., Rajkowska G., Stockmeier C.A., Krishtal O., Yakovleva T, & Bakalkin G. (2016). OPIOID PRECURSOR PROTEIN ISOFORM IS TARGETED TO THE CELL NUCLEI IN THE HUMAN BRAIN. Biochimica et biophysica acta, 1861(2), 246-255.

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In Vivo Fluorescence Imaging of Gastric Lesions

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Five 12-to-16-month-old IL1B mice were intravenously injected into the tail vein with 60 nmol of MK007 diluted in 0.9% NaCl (tot. vol. 150 µL; 400 nmol/mL) and sacrificed 4 h post injection (p.i.) by isoflurane overdosage. One non-injected 12-month-old IL1B mouse served as negative control. The whole stomach and esophagus were then excised and imaged using a homemade ex vivo wide-field fluorescence imaging system [15 (link)]. Briefly, the fluorescence images were acquired at different time exposure (0.2, 0.5, 1 and 2 s), using a 670 nm diode laser (B&W tek, Newark, DE, USA) and appropriate bandpass filter (780/10). A back illuminated EM-CCD camera (iXon DU888, Andor, Belfast, Northern Ireland, UK) was then used to capture the signal. The target-to-background ratio (TBR) of the stomach was performed by dividing the fluorescence signal in the squamocolumnar junction (SCJ) area (target lesion) with the adjacent normal gastric glandular tissue (background area) using ImageJ/Fiji software. In the organs, the signal quantification was performed by calculating the corrected total cell fluorescence/CTCF = integrated density – [(area * AVERAGE (mean background)]. For CTCF, three regions of interest (ROIs) without any tissue were used as background.

Marcazzan S., Braz Carvalho M.J., Konrad M., Strangmann J., Tenditnaya A., Baumeister T., Schmid R.M., Wester H.J., Ntziachristos V., Gorpas D., Wang T.C., Schottelius M, & Quante M. (2022). CXCR4 peptide-based fluorescence endoscopy in a mouse model of Barrett’s esophagus. EJNMMI Research, 12, 2.

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Visualization of Cellular Structures

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Viable cultures containing ALL cells and LEVs were immobilised onto CellTak coated glass coverslips, fixed with 3.7% paraformaldehyde probed using antibodies as detailed and imaged as described. LAMP1 and Talin: Super resolution microscopy was performed using a gSTED (gated Stimulated Emission Depleted) Leica TCS SP8 confocal microscope (Leica Biosystems, Wetzler, Germany) equipped with x63, NA1.4 oil lens, PMT and Hybrid (HyD) detectors, and with white laser (470–670 nm) and 405 UV laser. The software used was LAS X (Leica). Vinculin and time lapse images were captured using a Zeiss Axiovert 200 M microscope (Call Zeiss AG, Jena, Germany) fitted with a Zeiss_Plan-Fluor 0.5 numerical aperture connected to an Andor iXon DU888+ (Andor, Belfast, Northern Ireland) low light black and white camera. Illumination by UV light source was filtered using the SEDAT wheel filter set with appropriate wavelengths. The imaging system and image composites were achieved using Metamorph software (Molecular Devices, Sunnyvale, CA, USA).

Johnson S.M., Dempsey C., Parker C., Mironov A., Bradley H, & Saha V. (2017). Acute lymphoblastic leukaemia cells produce large extracellular vesicles containing organelles and an active cytoskeleton. Journal of Extracellular Vesicles, 6(1), 1294339.

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TIRF Imaging of Protein Adsorption

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TIRF microscopy was conducted to study protein adsorption using time-lapse imaging with fibrinogen from human plasma conjugated to Alexa Fluor 647 (Fisher). The time-lapse imaging was conducted using a custom-built microscope^{48 (link)} equipped with Olympus IX71 body, 100x objective and a CRISP ASI autofocus system. The excitation was conducted using a 638 nm laser (DL638–328 050, CrystaLaser, Reno, NV). Emission was collected using the appropriate Semrock bandpass filters and the images were acquired in a water-cooled, back-illuminated EMCCD camera (iXon DU-888, Andor, Belfast, UK) liquid-cooled to −70°C with an electronic gain of 60. An enzymatic oxygen scavenging system was used in the imaging buffer to reduce photobleaching.^{49 (link)} Imaging buffer consisted of 50 mM Tris-HCl (pH 8.0), 10 mM NaCl, 0.15 mg ml⁻¹ glucose oxidase, 34 μg ml⁻¹ catalase, 0.8% (w/v) glucose and 1% (v/v) β-mercaptoethanol. On SLIC surfaces, time-lapse imaging was conducted at a frame rate of 30 frames hr⁻¹. On other surfaces, the time-lapse imaging was conducted at a frame rate of 10 frames s⁻¹. During imaging, the concentration of fibrinogen was maintained at 5 nM for all experiments.

Vahabi H., Vallabhuneni S., Hedayati M., Wang W., Krapf D., Kipper M.J., Miljkovic N, & Kota A.K. (2022). Designing Non-Textured, All-Solid, Slippery Hydrophilic Surfaces. Matter, 5(12), 4502-4512.

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Visualizing HCV Infection Dynamics

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Huh7 YFP-TIA1 Neo cells (1.8 × 10⁴) with or without stable overexpression of PKR were seeded 24 hours before infection in 12-well plates with glass bottom (thickness 0.16 to 0.19 mm) (Cellvis). Cells were infected with HCV_TCP at an MOI of 1.5 TCID₅₀ per cell. Medium was replenished 24 hours after infection. Forty-eight hours after infection, culture medium was replenished with phenol red–free microscopy medium supplemented with IFN-α (100 IU/ml; PBL International) and transferred to the heating chamber of the microscope (Okolab). Image acquisition was performed using a Nikon Eclipse Ti2/Andor Revolution CSU-W1 spinning disc confocal microscope equipped with a 20× air objective CFI Plan Apo lambda [numerical aperture (NA) = 0.75], Nikon Perfect Focus System, Andor lasers 514 nm (40 mW) and 561 nm (50 mW), triple line dichroic beamsplitter 445/514/561, emission filters for YFP (540/30) and mCherry (600/50), Nikon motorized stage with linear encoder, electron multiplying charge-coupled device (EMCCD) Camera iXon DU-888—13 μm by 13 μm pixel size (Andor), and NIS Elements AR software (Nikon). Images (signals of YFP-TIA1 and NS5A-mCherry) were acquired in 15-min intervals for 72 hours, starting 4 to 5 hours after treatment with IFN-α. Typically, 15 to 30 fields of view were manually selected using the NS5A-mCherry signal and acquired simultaneously.

Klein P., Kallenberger S.M., Roth H., Roth K., Ly-Hartig T.B., Magg V., Aleš J., Talemi S.R., Qiang Y., Wolf S., Oleksiuk O., Kurilov R., Di Ventura B., Bartenschlager R., Eils R., Rohr K., Hamprecht F.A., Höfer T., Fackler O.T., Stoecklin G, & Ruggieri A. (2022). Temporal control of the integrated stress response by a stochastic molecular switch. Science Advances, 8(12), eabk2022.

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Live-cell and Fixed-cell Imaging Protocol

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Live cells and fixed cells were imaged on an inverted microscope (Ti Eclipse; Nikon) at room temperature using a 40× NA 0.6 Plan Fluor air or a 100× NA 1.4 Plan-Apochromat oil objective lens (Nikon). All images were collected with a cooled back-thinned EM charge-coupled device (CCD) camera (iXon+ DU888; Andor Technology) with a 1× or 1.5× optivar using MicroManager 1.4 software. Live cells were imaged in their usual culture media. For cell tracks, a randomly selected region spanning a tiled series of 10 × 10 or 6 × 6 fields of view was imaged every minute for 20 min. For drug perturbation experiments, cells were imaged every 10 s for 10 min before drug addition, and continuing for 30 min after drug addition. For live-cell myosin experiments, cells were imaged every 3 s. Fixed cells were mounted in Vectashield mounting media (Vector Laboratories).
For 3D structured illumination microscopy experiments, fixed cells were mounted in SlowFade Gold mounting media (Invitrogen) and imaged on a DeltaVision OMX (GE Healthcare) microscope using DeltaVision software (GE Healthcare) with a 100× NA 1.4 UPlan-Apochromat oil objective lens (Olympus) using 0.125 µm z slices at room temperature. Images were collected with a cooled back-thinned EM-CCD camera (Evolve; Photometrics). 3D structured illumination reconstructions were created with the softWoRx imaging suite (GE Healthcare).

Lou S.S., Diz-Muñoz A., Weiner O.D., Fletcher D.A, & Theriot J.A. (2015). Myosin light chain kinase regulates cell polarization independently of membrane tension or Rho kinase. The Journal of Cell Biology, 209(2), 275-288.

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Ex vivo Fluorescence Imaging of Hsp70-TPP-Cy5.5

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To verify the in vivo biding capabilities of Hsp70-TPP-Cy5.5 to target malignant lesions and to validate the fluorescence molecular endoscopy, we used an ex vivo fluorescence imaging system to detect Hsp70-TPP-Cy5.5-derived fluorescence signals on exposed or excised specimen. For near-infrared epifluorescence imaging in tissue, specimens were illuminated using a 670-nm diode laser (B&W tek, DE, USA). The emitted fluorescence was guided through a 780/10 bandpass filter and captured with a back-illuminated EM-CCD camera (iXon DU888, Andor), as described previously [27 (link)]. Fluorescence images were sequentially acquired at exposure times of 0.2, 0.5, 1, and 2 s. Comparative analysis has been carried out at images taken with identical exposure times, objective apertures, and camera settings. Signal specificity was determined by calculating the ratio of the mean signal intensities of the relevant tissue and the adjacent normal tissue using ImageJ as previously reported [52 (link)]. Data are represented as single plotted values and/or mean ± SEM. Imaging procedures of the different types of specimen have been performed as follows.

Fang H.Y., Stangl S., Marcazzan S., Carvalho M.J., Baumeister T., Anand A., Strangmann J., Huspenina J.S., Wang T.C., Schmid R.M., Feith M., Friess H., Ntziachristos V., Multhoff G., Gorpas D, & Quante M. (2021). Targeted Hsp70 fluorescence molecular endoscopy detects dysplasia in Barrett’s esophagus. European Journal of Nuclear Medicine and Molecular Imaging, 49(6), 2049-2063.

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Spinning Disc Confocal Microscopy

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Spinning disc confocal microscopy (SDCM) was performed on an inverted Nikon Eclipse Ti2 (Nikon, Japan) microscope equipped with a Yokogawa CSU-W1 Spinning Disk Unit (Andor, Oxford Instruments, United Kingdom) and an incubation chamber (37°C, 5% CO₂). Imaging was performed using a 100 × oil immersion objective (Nikon CFI Apochromat TIRF 100X Oil NA 1.49) and either single or dual-channel EMCCD camera setup (ANDOR iXon DU-888) recording the eBFP2 (405/420–460), eGFP (488/510–540 nm), mScarlet (568/589–625 nm) and SiR (647/665–705 nm) channels with a pixel size of 0.13 µm. 3D stacks of 10–30 randomly chosen positions were automatically recorded with a z-spacing of 0.3–0.5 µm using the Nikon Imaging Software Elements v5.02. For CLEM experiments a Perkin Elmer Ultra VIEW VoX 3D spinning disk confocal microscope (Perkin Elmer, United States) with a 100 x oil immersion objective (NA 1.4; Perkin Elmer) was used.

Müller T.G., Zila V., Peters K., Schifferdecker S., Stanic M., Lucic B., Laketa V., Lusic M., Müller B, & Kräusslich H.G. (2021). HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells. eLife, 10, e64776.

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