HeLa cells were plated on 96-well glass bottom plates (Matriplate) at the density of 8 × 103 ∼ 1 × 104 per well. Transient transfection was performed with Fugene HD (Promega) 24 hrs later according to manufacturer’s protocol. A mixture of plasmids comprising SunTag system with equal amount (50 ng) for each was first made in Opti-MEM then Fugene HD was added and incubated for 15 mins at room temperature. The amount of DNA and Fugene HD can be optimized accordingly. 24 hrs after transfection cells were imaged using a DeltaVision Elite microscope with a 60× objective (1.42 NA; Olympus) and temperature-matched oil in an environmentally controlled chamber (37°C, 5% CO2). Z-stacks of 5 images with 1 μm interval were acquired for quantification with dNEMO. Cell segmentation was manually performed in dNEMO and the spot detection parameters are set as default. In comparison of SunRISER and MS2-MCP, stacks of 4 planes with a z-spacing of 0.5 μm were obtained for high frame rate (one 3D stack per 2s). For comparison of promoters, mean fluorescence intensity was measured using ImageJ for a fixed region in cytoplasmic area.
Deltavision elite
Manufactured by Olympus
Sourced in United States, Japan, Germany
The DeltaVision Elite is a high-performance microscope system designed for advanced imaging applications. It features an inverted optical configuration and supports a range of sample types, including live-cell, fixed-cell, and 3D samples. The system is equipped with advanced optics, a motorized stage, and a high-sensitivity camera, enabling high-resolution, high-contrast imaging. The DeltaVision Elite is a versatile tool for researchers in various fields, such as cell biology, developmental biology, and materials science.
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Lab products found in correlation
Ix71 stand, by Olympus (6 mentions)
Ix71 inverted microscope, by Olympus (3 mentions)
Deltavision elite system, by GE Healthcare (3 mentions)
Ultimate focus, by Olympus (2 mentions)
Fugene hd, by Promega (1 mentions)
Softworx software, by GE Healthcare (1 mentions)
Planapo n 60, by Olympus (1 mentions)
Oil super plan apo chromatic objective, by Olympus (1 mentions)
Lysotracker red, by Thermo Fisher Scientific (1 mentions)
Softworx, by Cytiva (1 mentions)
Deltavision software, by GE Healthcare (1 mentions)
Texas red, by Vector Laboratories (1 mentions)
Cy5 conjugated secondary antibodies, by Abcam (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Axiovert 200m, by Zeiss (1 mentions)
384 well non binding microplates, by Greiner (1 mentions)
Plan apochromat vc 60x 1.2 na water objective, by Nikon (1 mentions)
Low melting agarose, by Merck Group (1 mentions)
Uplansapo 100 1.40 oil objective, by Olympus (1 mentions)
Fm4 64, by Thermo Fisher Scientific (1 mentions)
21 protocols using deltavision elite
1
SunTag for Live-Cell Imaging of RNA
2
Visualizing PGRP-SA and PGRP-LC Binding
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Bacteria were grown to OD600nm ∼0.5 and 1 mL aliquot was centrifuged for 5 min at RT, 16.1 x1000 g. The cells were washed with 500 μL of HemoBuffer and centrifuged again. They were resuspended in 0.3 mg/mL final concentration of either mCherry_PGRP-SA or mCherry_PGRP-LC in a 200 μL reaction volume, filling up the volume with Hemo buffer and incubated at RT for 5 min without shaking. The “cells-bound PGRPs” complexes were harvested for 5 min at RT, 7.5 x1000 g. The pellets were washed twice with 200 μL of Buffer. Finally, the pellets were resuspended in the left-over volume of the washings and 2 μL was loaded on a 1.2% (w/v) Agarose-PBS slides (it does not dissolve in HemoBuffer). Images were acquired with a GE Healthcare DeltaVision Elite integrated imaging system, in the conventional mode at 25°C using an Olympus 150x 1.45 NA TIRF Objective (Olympus, USA) using the SoftWoRx software (GE Healthcare).
3
High-Resolution Imaging of Fluorescent Cells
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High-resolution imaging was performed using a General Electric (Boston, MA) DeltaVision Elite imaging system mounted on an Olympus (Japan) IX71 stand with a computerized stage, Ultimate Focus, environment chamber, ultrafast solid-state illumination with excitation/emission filter sets for CFP, GFP, YFP, mCherry, and Cy5, critical illumination, Olympus PlanApo N 60X/1.42 NA DIC (oil) and UPlanSApo 60X/1.3 NA DIC (sil) objectives, Photometrics (Tucson, AZ) CoolSNAP HQ2 camera, proprietary constrained iterative deconvolution, and vibration isolation table.
4
High-Resolution Imaging System Protocol
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High-resolution imaging was performed using a General Electric (Boston, MA) DeltaVision Elite imaging system mounted on an Olympus (Japan) IX71 stand with a motorized XYZ stage, Ultimate Focus, cage incubator, ultrafast solid-state illumination with excitation/emission filter sets for DAPI, CFP, FITC, GFP, YFP, TRITC, mCherry, and Cy5, critical illumination, Olympus PlanApo N 60 ×/1.42 NA DIC (oil) and UPlanSApo 60 ×/1.3 NA DIC (silicone) objectives, Photometrics (Tucson, AZ) CoolSNAP HQ2 camera, SoftWoRx software with constrained iterative deconvolution, and vibration isolation table.
5
High-Resolution Live Cell Imaging
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High resolution live cell imaging was performed using an Olympus-based Delta Vision Elite work station fitted with a 100x objective (numerical NA 1.4, oil). Post-acquisition analysis was carried out using Applied Precision software. Images presented are 2D projections of deconvolved Z-stacks of 0.3 μm optical sections.
6
High-Resolution Live Cell Imaging
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High resolution live cell imaging was performed using an Olympus-based Delta Vision Elite work station fitted with a 100× objective (numerical NA 1.4, oil). Post-acquisition analysis was carried out using Applied Precision software. Images presented are 2D projections of deconvoluted Z-stacks of 0.3 μm optical sections.
7
In vivo Ypq1-GFP Localization Assay
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For in vivo experiments, Ypq1-GFP was expressed under its native promoter in yeast auxotrophic to multiple amino acids (YML1415). Cells were grown in rich media (YPD) in 26°C overnight with shaking. The next day, cell pellets were centrifuged at 13,500 × g for 1 min, washed twice with MQ H2O, transferred to YNB media lacking the test amino acid, and incubated at 26°C for 6 h with shaking. At time intervals, cells were collected for 1 min at 13,500 × g, washed with MQ H2O, and imaged immediately.
Microscopy was performed with a DeltaVision Elite system (GE Healthcare Life Sciences), equipped with an Olympus IX-71 inverted microscope, a scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera, a 100 × /1.4 Oil Super-Plan Apo-chromatic objective, and a DeltaVision Elite Standard Filter Set with the FITC filter (Excitation:475/28, Emission:525/48) for GFP. Image acquisition and deconvolution were performed in the program Softworx. ImageJ (NIH) was used for image processing.
Microscopy was performed with a DeltaVision Elite system (GE Healthcare Life Sciences), equipped with an Olympus IX-71 inverted microscope, a scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera, a 100 × /1.4 Oil Super-Plan Apo-chromatic objective, and a DeltaVision Elite Standard Filter Set with the FITC filter (Excitation:475/28, Emission:525/48) for GFP. Image acquisition and deconvolution were performed in the program Softworx. ImageJ (NIH) was used for image processing.
8
Yeast Cell Microscopy Using DeltaVision Elite
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Microscopy was performed with a DeltaVision Elite system (GE Healthcare Life Sciences), equipped with an Olympus IX-71 inverted microscope, a sCMOS camera, a 100X/1.4 Oil Super-Plan APO objective, and a DeltaVision Elite Standard Filter Set with the FITC filter (Excitation:475/28, Emission: 525/48) for mNeonGreen and the TRITC filter (Excitation:542/27, Emission: 594/45) for mCherry, DsRed, and FM4-64. Yeast cells, except for those transformed with the pRS416-DsRed-HDEL plasmid, were grown in YPD at 26°C overnight. Yeast cells transformed with the pRS416-DsRed-HDEL plasmid were grown in YNB minus uracil at 26°C overnight. Before imaging, yeast cells were briefly washed with water and immediately imaged in milliQ water at room temperature. Image acquisition, deconvolution, and maximum projection analysis were performed with the program softWoRx. The image cropping and adjustment were performed using the ImageJ software (National Institutes of Health).
9
Monitoring Amino Acid-Dependent Ypq1-GFP Localization
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For in vivo experiments, Ypq1-GFP was expressed under its native promoter in yeast auxotrophic to multiple amino acids (YML1415). Cells were grown in rich media (YPD) in 26°C overnight with shaking. The next day, cell pellets were centrifuged at 13,500xg for 1 min, washed twice with MQ H2O, transferred to YNB media lacking the test amino acid, and incubated at 26°C for 6 hrs with shaking. At time intervals, cells were collected for 1 min at 13,500xg, washed with MQ H2O, and imaged immediately.
Microscopy was performed with a DeltaVision Elite system (GE Healthcare Life Sciences), equipped with an Olympus IX-71 inverted microscope, a scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera, a 100×/1.4 Oil Super-Plan Apo-chromatic objective, and a DeltaVision Elite Standard Filter Set with the FITC filter (Excitation:475/28, Emission:525/48) for GFP. Image acquisition and deconvolution were performed in the program Softworx. ImageJ (NIH) was used for image processing.
Microscopy was performed with a DeltaVision Elite system (GE Healthcare Life Sciences), equipped with an Olympus IX-71 inverted microscope, a scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera, a 100×/1.4 Oil Super-Plan Apo-chromatic objective, and a DeltaVision Elite Standard Filter Set with the FITC filter (Excitation:475/28, Emission:525/48) for GFP. Image acquisition and deconvolution were performed in the program Softworx. ImageJ (NIH) was used for image processing.
10
High-Resolution Live Cell Imaging
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Live high-resolution imaging was performed using a General Electric (Boston, MA) DeltaVision Elite imaging system mounted on an Olympus (Japan) IX71 stand with a computerized stage, Ultimate Focus, environment chamber (heat, CO2, and humidifier), ultrafast solid-state illumination with excitation/emission filter sets for DAPI, CFP, GFP, YFP, and Cy5, critical illumination, Olympus PlanApo N 60X/1.42 NA DIC (oil), UPlanSApo 20X/0.75 NA DIC (air), and UPLSAPO 10X/0.4 NA DIC (air) objectives, Photometrics (Tucson, AZ) CoolSNAP HQ2 camera, proprietary constrained iterative deconvolution, and vibration isolation table.
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