Du885

Manufactured by Oxford Instruments

Sourced in United States

The DU885 is a laboratory instrument designed for ultraviolet-visible (UV-Vis) spectroscopy. It is capable of measuring the absorbance or transmittance of light in the ultraviolet and visible wavelength ranges.

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Lab products found in correlation

Elyra ps1 system, by Zeiss (2 mentions) Eclipse ti e, by Nikon (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Rhod 2 am, by Cayman Chemical (1 mentions) Polyfect transfection reagent, by Qiagen (1 mentions) Acetylated α tubulin, by Santa Cruz Biotechnology (1 mentions) Zen black software, by Zeiss (1 mentions) Plan apo vc 100 oil dic n2, by Nikon (1 mentions) Nis elements ar 4, by Nikon (1 mentions) Na 1.40 oil immersion objective, by Nikon (1 mentions) Cfi plan apo vc60xh objective, by Nikon (1 mentions) Swept field confocal, by Nikon (1 mentions) Ti microscope, by Nikon (1 mentions) Zen 2012, by Zeiss (1 mentions) Eclipse ti e microscope, by Nikon (1 mentions) Cellasic onix system, by Merck Group (1 mentions)

Spelling variants of this product

DU885 EM-CCD camera (10 citations) DU885 electron-multiplying CCD camera (5 citations) DU885 EMCCD (3 citations)

9 protocols using du885

Visualizing Actin and Calcium Dynamics

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For the observation of intracellular actin in live cells, we utilized PolyFect Transfection Reagent (QIAGEN, Hilden, Germany) to transfect the cells with the pTagRFP-actin vector (Evrogen, Moscow, Russia). The exposure time for an actin image was 1.0 s. For the observation of intracellular calcium ions in live cells, we used the calcium ion indicator Rhod-2 AM (Cayman Chemical, Ann Arbor, MI, USA) to stain the cells for 1 h, and then replaced the medium. The exposure time for a calcium image was 1.0 s. We set the electron-multiplying gain of the camera as 10 for acquiring the calcium images. During the capture of each actin or calcium image, the red light was temporarily blocked for ~30 s. The average pixel value (after background subtraction) within a cell normalized to that of the same cell measured before any optical stimulation was recorded as the relative intracellular calcium level.
The fluorescence images were acquired using an epifluorescence microscope (Eclipse Ti-E, Nikon, Tokyo, Japan). The images were captured by a 14-bit electron-multiplying charge-coupled device (EMCCD) camera (DU-885, Andor, Belfast, Northern Ireland) cooled down to −60 °C. The fluorescence intensity of individual cells was quantified using ImageJ (http://rsb.info.nih.gov/ij/).

Kao Y.C., Liao Y.C., Cheng P.L, & Lee C.H. (2019). Neurite regrowth stimulation by a red-light spot focused on the neuronal cell soma following blue light-induced retraction. Scientific Reports, 9, 18210.

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Quantitative Super-Resolution Imaging of Acetylated Microtubules

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Transfected cells were seeded into 6-well dishes containing 5 mm round coverslips and 2 mL fresh media treated with chloroquine after 24 hours, followed by fixation for 10 minutes in a 1:1 ratio of 4% Paraformaldehyde: DMEM at 37°C. Cells were blocked using 3% donkey serum in PBS, followed by probing the required primary antibody, i.e. acetylated α-tubulin (Santa Cruz, 23950) overnight at 4°C. Next, cells were incubated with a secondary antibody, Alexa Fluor 568 (ThermoFisher, A-10042) for 50 minutes. Coverslips were then washed 3x5 minutes with PBS and using Dako^® fluorescent mounting media. For SR-SIM analysis of GFP-Tau transfected cells, thin (0.1 μm) z-stacks of high-resolution (1024x1024 pixels) image frames were collected by utilizing an alpha Plan-Apochromat 60x/1.4 oil immersion DIC M27 ELYRA objective on an ELYRA PS.1 system (Carl Zeiss Microimaging, Germany) equipped with a 488nm laser (100mW) in 5 rotations, 561nm laser (100mW) in 5 rotations and an Andor EM-CCD camera (iXon DU 885). Images were reconstructed using Zeiss Zen Black Software (2012) based on a structured illumination algorithm [22 ]. Colocalization analysis of these reconstructed super-resolution images using both the 2D and 3D VR-assisted 3D ROI selection systems.

Theart R.P., Loos B., Powrie Y.S, & Niesler T.R. (2018). Improved region of interest selection and colocalization analysis in three-dimensional fluorescence microscopy samples using virtual reality. PLoS ONE, 13(8), e0201965.

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Live-cell Imaging of Transfected HeLa Cells

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HeLa cells were analyzed 24–48 h after transfection using an inverted Nikon Eclipse Ti microscope equipped with a 100× objective (Plan Apo VC 100× Oil DIC N2; Nikon). EGFP and mCherry/DsRed channels were acquired simultaneously using a dual camera port system composed of custom-made excitation/emission filters, a dual-band beam splitter, and two cooled 14-bit electron-multiplying CCD cameras (DU-885; Andor Technology, Belfast, UK). For recording live-cell image sequences, cells were maintained at 37°C by means of a stage heater (ibidi, Martinsried, Germany) with a temperature control system (TC 20; npi, Tamm, Germany) and an objective heater (PeCon, Erbach, Germany). Images and sequences were acquired and analyzed with NIS Elements AR 4 software (Nikon).

Renigunta V., Fischer T., Zuzarte M., Kling S., Zou X., Siebert K., Limberg M.M., Rinné S., Decher N., Schlichthörl G, & Daut J. (2014). Cooperative endocytosis of the endosomal SNARE protein syntaxin-8 and the potassium channel TASK-1. Molecular Biology of the Cell, 25(12), 1877-1891.

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Imaging Bacterial Cells Under Microfluidic Flow

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Overnight cultures were diluted 1:200 and grown at 37 °C, followed by two 1:10 dilutions (at OD₆₀₀ around 0.1) to generate steady state exponential phase cells. Cells were imaged on a Nikon Eclipse Ti-E inverted fluorescence microscope with a 100X (NA 1.40) oil-immersion objective (Nikon Instruments). Images were collected on a DU885 electron-multiplying charged couple device camera (Andor Technology) using μManager version 1.4 (34 ). Cells were maintained at 37 °C during imaging with an active-control environmental chamber (HaisonTech).
For single-cell imaging on agarose pads, 1 μL of cells was spotted onto a pad of 1% agarose in fresh LB. For microfluidic flow-cell experiments, including the spent media assays, the oscillatory osmotic shock assays, and the plasmolysis/lysis assays, cells were loaded to CellASIC ONIX microfluidic chips (Sigma-Aldrich, Cat. #B04A-03-5PK) and medium was exchanged using the CellASIC ONIX2 microfluidic platform (Sigma-Aldrich, Cat. #CAX2-S0000).

Mikheyeva I.V., Sun J., Huang K.C, & Silhavy T.J. (2023). Mechanism of outer membrane destabilization by global reduction of protein content. bioRxiv.

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Imaging of HeLa cells expressing C19orf12

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HeLa cells were co-transfected with GFP-tagged C19orf12 wild-type or mutant chimeras and the mitochondrial marker mtDsRed using calcium phosphate method. Thirty six hour after transfection, time-lapse recording were performed with a Nikon Swept Field Confocal equipped with CFI Plan Apo VC60XH objective (numerical aperture, 1.4) (Nikon Instruments, Melville, NY, USA) and an Andor DU885 electron multiplying charge- coupled device (EM-CCD) camera (Andor Technology Ltd, Belfast, Northern Ireland), the overall image sampling was below the resolution limit (X and Y pixel size: 133 nm). Coverslips were placed in an incubated chamber with controlled temperature, CO₂ and humidity; images were then acquired with a differential frequency during the experiment: cells were placed in 1 mM Ca²⁺ KRB and basal fluorescence images were acquired for 5 min; then cells were stimulated with H₂O₂ (500 μM final), and fluorescence images were acquired for 1 h and 30 min.

Venco P., Bonora M., Giorgi C., Papaleo E., Iuso A., Prokisch H., Pinton P, & Tiranti V. (2015). Mutations of C19orf12, coding for a transmembrane glycine zipper containing mitochondrial protein, cause mis-localization of the protein, inability to respond to oxidative stress and increased mitochondrial Ca2+. Frontiers in Genetics, 6, 185.

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Live cell RUSH imaging protocol

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A Nikon Ti microscope with a Plan Apo VC 60× oil differential interference contrast (DIC) lens and an Andor DU-885 camera were used to acquire live wide-field RUSH images. The heated stage was pre-heated to 37°C. The cell medium was replaced with 1 mL pre-warmed HBS. When RUSH-transfected cells were found, biotin was added to the cells by diluting biotin to a 2× working solution in HBS. 1 mL of this 2× biotin working solution was added to the original 1 mL of imaging medium already on the cells. The cells were imaged over time periods of up to 60 min, with a frame being taken every 5 or 10 s.

Evans A.J., Gurung S., Wilkinson K.A., Stephens D.J, & Henley J.M. (2017). Assembly, Secretory Pathway Trafficking, and Surface Delivery of Kainate Receptors Is Regulated by Neuronal Activity. Cell Reports, 19(12), 2613-2626.

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Fluorescence Microscopy for Live-Cell Imaging

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Cells were imaged on a Nikon Eclipse Ti-E inverted fluorescence microscope with a 100× oil-immersion lens objective (Nikon Instruments) (numerical aperture, 1.40). Images were collected on a DU885 electron-multiplying charge-coupled-device (CCD) camera (Andor Technology) or a Neo scientific complementary metal oxide semiconductor (sCMOS) camera (Andor Technology) using μManager v. 1.4 (50 (link)). Cells were maintained at 37°C during imaging with an active-control environmental chamber (Haison Technology). One microliter of cells was spotted onto a pad of 1% agarose plus medium as noted.

Cesar S., Anjur-Dietrich M., Yu B., Li E., Rojas E., Neff N., Cooper T.F, & Huang K.C. (2020). Bacterial Evolution in High-Osmolarity Environments. mBio, 11(4), e01191-20.

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Structured Illumination Imaging Protocols

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Structured illumination imaging was performed using a Zeiss Elyra PS1 system. 3D-SIM data was acquired using a 63 × 1.4 NA oil objective. 488, 561, 642 nm 10 0 mW diode lasers were used to excite the fluorophores together with, respectively, a BP 495–575+LP 750, BP 570–650+LP 750 or LP 655 excitation filter. For 3D-SIM imaging the recommended grating was present in the light path. The grating was modulated in 5 phases and 5 rotations, and multiple z-slices with an interval of 110 nm were recorded on an Andor iXon DU 885, 1,002 × 1,004 EMCCD camera. Raw images were reconstructed using the Zeiss Zen 2012 software. For SIM representative images, the following number of experiments were performed: (i) glass versus scratches, three experiments, per experiment five images from different cells for each condition; (ii) CytoD versus untreated, three experiments, per experiment five images from different cells for each condition; (iii) myosin IIA inhibition versus untreated: two experiments (one with Blebb and one with ML7+Y27632 mix), five images from different cells for each condition.

Meddens M.B., Pandzic E., Slotman J.A., Guillet D., Joosten B., Mennens S., Paardekooper L.M., Houtsmuller A.B., van den Dries K., Wiseman P.W, & Cambi A. (2016). Actomyosin-dependent dynamic spatial patterns of cytoskeletal components drive mesoscale podosome organization. Nature Communications, 7, 13127.

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Live Cell Permeability Imaging

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Imaging was performed on a Ti-E Eclipse microscope (Nikon Instruments, Inc., Melville, NY, USA) with a 1.4NA 100X objective. Images were acquired with an Andor DU885 (Andor Technology, South Windsor, CT, USA) camera, using μManager v. 1.4.⁴⁴ Temperature was maintained within a custom environmental chamber (HaisonTech, Taipei, Taiwan). To measure cell permeability after treatment, cells were loaded into a B04 bacterial microfluidic chip in a CellASIC ONIX system (EMD Millipore, Hayward, CA, USA) and propidium iodide fluorescence was monitored every 1 min.

Ochoa J.L., Sanchez L.M., Koo B.M., Doherty J.S., Rajendram M., Huang K.C., Gross C.A, & Linington R.G. (2017). Marine Mammal Microbiota Yields Novel Antibiotic with Potent Activity Against Clostridium difficile. ACS infectious diseases, 4(1), 59-67.

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