Cells were plated on glass-bottom MatTek dishes coated with fibronectin (1 µg ml−1) and maintained in complete media in a 37 °C incubator with 5% CO2. The next day, the media was changed to Leibovitz’s L-15 (Gibco)/10% serum or FluoroBrite DMEM (Gibco)/10% serum depending on the CO2 microscope environment. Live imaging of mCherry-CaaX/pHluo_M153R-CD63-expressing HT1080 cells was performed with a Nikon Eclipse TE2000E epifluorescence microscope (ran by MetaMorph software, Molecular Devices) equipped with a 37 °C chamber and a cooled CCD camera using a Nikon Plan Fluor oil 40x/1.30 NA or Super Fluor 20x/0.75 NA objective lens. Still images of pHluo_M153R-CD63 were captured from diverse live cells using a Nikon Plan Fluor oil 40x/1.30 NA lens on the Nikon Eclipse TE2000E (ran by MetaMorph software, Molecular Devices). Time-lapse movies of pHluo_M153R-CD63-expressing cells on nanopatterned glass-bottom dishes (800 nm width of both ridge and groove and 600 nm depth, Nanosurface Biomedical) coated with fibronectin (1 µg ml−1), and of cells expressing the dual reporter pHluo_M153R-CD63-mScarlet, were acquired with a Nikon A1R confocal microscope (ran by NIS-Elements) equipped with a Tokai Hit Incubation Chamber (37 °C with 5% CO2) using a Plan Apo 40x/1.3 NA oil immersion lens.
Eclipse te2000 e
Manufactured by Nikon
Sourced in Japan, United States, Germany, France
The Eclipse TE2000-E is a high-performance inverted microscope designed for advanced cell and tissue culture applications. It features a sturdy, ergonomic design and a range of optical configurations to support a variety of research requirements. The Eclipse TE2000-E provides researchers with the tools necessary to conduct detailed analysis and observation of biological samples.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (9 mentions)
Hoechst 33342, by Thermo Fisher Scientific (7 mentions)
Nis element, by Nikon (5 mentions)
Metamorph software, by Molecular Devices (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions)
Orca flash 4, by Hamamatsu Photonics (4 mentions)
Ez c1, by Nikon (4 mentions)
Rabbit anti iba1, by Fujifilm (3 mentions)
Dpx mounting media, by Merck Group (3 mentions)
Tunel assay, by Roche (3 mentions)
Nis elements ar software, by Nikon (3 mentions)
Eclipse c1 confocal microscope, by Nikon (3 mentions)
Paraformaldehyde (pfa), by Merck Group (3 mentions)
L3224, by Thermo Fisher Scientific (3 mentions)
Rabbit anti myeloperoxidase mpo, by Agilent Technologies (2 mentions)
Alexa 488 conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 or 568 labeled secondary antibody, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Roche (2 mentions)
25mm glass coverslips, by Thermo Fisher Scientific (2 mentions)
Fura 2 am, by Thermo Fisher Scientific (2 mentions)
242 protocols using eclipse te2000 e
1
Visualizing Endosomal Dynamics in 3D
2
Macrophage-Streptococcus Interaction Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in Delta T culture dish (Bioptechs) at a density of 4 × 105 cells/ml and differentiated into macrophages by incubation with 100 nM PMA. Based on the results of previous experiments, three different GBS isolates were selected: isolate 231 (serotype III, ST-17), isolate 9427 (serotype Ia, ST-144) and isolate 203 (serotype Ib, ST-8). Bacteria were prepared as described in 2.3 and 2.4 and added to macrophages at a MOI of 10. The Delta T culture dish was placed in a confocal microscope (Nikon eclipse TE2000-E) and heated to 37°C throughout the experiment. In 3 h, 360 images were acquired, i.e., 1 image per 30 s with 600x magnification.
3
Transwell Assay for Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell migration was detected by a Transwell assay [25 (link)]. Briefly, hUC-MSCs treated with rhMG53 (30 μg/ml) and/or H2O2 (200 μM, 16 h) were collected and resuspended in DMEM/F12 media without serum, and re-plated in transwell with 8.0 μm holes at a density of 1 × 105 cells/ml, 300 μl/well. Twenty-four hours later, the cells were fixed with 4% paraformaldehyde for 10 min and stained with crystal violet for 30 min. The cells in the upper chamber were removed, and the migrated cells were detected as purple in color and counted under a microscope (Eclipse TE2000-E; Nikon, Japan).
4
Fluorescent Protein Localization Assay
5
Flow Cytometric Analysis of DC-Tumor Fusion Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Patient-derived breast carcinoma cells and patient-derived DCs were incubated with primary mouse anti-human mAbs against the tumor antigens MUC1, HER2, HLA-A2, HLA-A11, HLA-DR and CD86 and matched isotype controls (BD Pharmingen). Cells were then washed, and incubated with FITC-conjugated mouse anti-mouse IgG1 (BD Pharmingen). DC-tumor fusion cell preparations were subjected to dual staining to quantify the percentage of cells that co-expressed the unique DC marker (HLA-DR-PE) and the tumor marker (HER2-FITC) (BD Pharmingen). Cells were fixed in 2% paraformaldehyde (Sigma-Aldrich) and underwent flow cytometry analysis using FACScan (BD Biosciences) and CellQuest software (BD Biosciences). True fusion cells were identified by immunofluorescence staining and laser confocal microscopy. Approximately 2×104 DC-tumor fusion cells were spun onto slides (Cytospin 4 Thermo Shandon), allowed to dry, and fixed with 4% paraformaldehyde for 8 min. The slides were incubated with FITC-conjugated mouse anti-human HER2 mAb for 1 h at 4°C. The slides were then washed and incubated with PE-conjugated mouse anti-human HLA-DR mAb for 1 h at 4°C before being washed, fixed in 4% paraformaldehyde, and analyzed by confocal microscopy (Nikon Eclipse TE2000-E).
6
Immunofluorescence of Cellular Markers in ICH
7
Time-Lapse Migration Assay for Primary Fibroblasts
8
Immunofluorescence Analysis of ZIKV Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed in 4% paraformaldehyde and stained with the primary antibodies (Supplementary Table 4 ) after permeabilization with 0.1% Triton X-100/0.5% BSA/PBS solution. Appropriate Alexa Fluor® 488 or 568 labeled secondary antibody (Life Technologies) and DAPI (Roche Applied Science) nuclear counter-staining were used for visualization. The stained samples were analyzed using fluorescence microscopy (Eclipse TE2000-E, Nikon). The numbers of ZIKVE+, Ki-67+, or DAPI+ cells per mm2 were counted under fluorescence microscopy. Cell viability 65 h after ZIKVPR infection was represented as percentage fold change (relative to mock) of DAPI+ cell number (/mm2) in ZIKVPR-infected cells (mean ± s.e.m.). The experimenter was not blinded to treatment. None of cell cultures were excluded from our analyses.
9
Immunofluorescence Assay for EBNA1 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The day before, HEK293T cells stably expressing the EBNA1-FLAG-HA protein were seeded at a density of 5 × 104 cells/well in 24-well plates. Cells were washed twice with PBS and fixed 20 min using 4% paraformaldehyde and 4% sucrose in PBS at room temperature. Cells were then permeabilized with 0.15% triton X-100 in PBS for 5 min at room temperature and blocked in 10% normal goat serum (Wisent). Anti-HA antibody (Santa Cruz Biotechnologies) was incubated 4 h at room temperature to allow detection of EBNA1-FLAG-HA protein. Cells were washed and incubated 1 h in the dark with DyLight 488-labelled goat anti-mouse secondary antibody (ThermoFisher Scientific). Nucleus staining was performed using 1 μg/ml Hoechst, 15 min at room temperature. Cover glasses were mounted on slides with SlowFadeGold mounting medium (Life Technologies), then epifluorescence microscopy was conducted using a Nikon Eclipse TE2000-E visible/epifluorescence inverted microscope using bandpass filters for Hoechst and DyLight 488.
10
Quantifying H3N2 Antigen Diffusion in Skin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To quantify the diffusion profile of H3N2 in the skin over time, FITC-dextran (MW: 70 kDa) was used as a model drug with a molecular weight similar to that of the H3N2 antigen (MW: 75 kDa). At 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, and 330 min after microneedle administration, the distribution of fluorescence of FITC-dextran in the skin was observed using a confocal microscope (Ex: 490 nm/Em:520 nm, ECLIPSE TE2000-E, Nikon, Tokyo, Japan). Sequential z-stack images were captured at 15-μm-depth intervals from the skin surface to a depth of 840 μm. Analysis was performed using a plot profile of Image J (National Institutes of Health, Bethesda, MD) of fluorescent signal intensity in confocal z-stacks obtained along the inserted microneedles.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!