HeLa cells (5 × 104) were seeded on cover glass in 24-well plates and transfected with 0.3 μg of pNL43 Luc E- R+ or pNL43 Luc E- R-, either without/with 0.1 μg of pCAGGS/mRFP-TSG101 and 0.1 μg of pcDNA/YFP-Vpr plasmids, for 16 h. The cells were then fixed in 3.7% formaldehyde, permeabilized with 0.5% Triton X100, and stained with anti-p24 mouse mAb AG3.0 [33 (link)] and FITC-conjugated sheep anti-mouse Fab’ (ICN Biomedicals). TIRF images were acquired by an Eclipse TE2000-E inverted microscope (Nikon).
Eclipse te2000 e inverted microscope
Manufactured by Nikon
Sourced in Japan, United States
The Eclipse TE2000-E is an inverted microscope designed for a variety of laboratory applications. It features a sturdy, ergonomic design and provides high-quality imaging capabilities. The microscope is equipped with a range of optical components and can accommodate various sample types. Its core function is to enable detailed observation and analysis of specimens under controlled laboratory conditions.
Automatically generated - may contain errors
Lab products found in correlation
Ez c1, by Nikon (3 mentions)
Odyssey 9120 imager, by LI COR (2 mentions)
Image studio software, by LI COR (2 mentions)
H 1200, by Vector Laboratories (2 mentions)
Dark field oil condenser, by Nikon (2 mentions)
S fluor 20 0.75 air objective, by Nikon (2 mentions)
Prism 7, by GraphPad (2 mentions)
Actin stain 670 phalloidin, by Tebu-Bio (2 mentions)
Nucgreen dead 488, by Thermo Fisher Scientific (2 mentions)
Metamorph software, by Molecular Devices (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Quanta 600, by Thermo Fisher Scientific (1 mentions)
35 mm glass bottom imaging dishes, by MatTek (1 mentions)
Spectra x led light engine, by Lumencor (1 mentions)
Clara charge coupled device camera, by Oxford Instruments (1 mentions)
Fluorodish cell culture dishes, by World Precision Instruments (1 mentions)
Di01 r405 488 561 635 25x36, by IDEX Corporation (1 mentions)
Superk extreme exw 12, by NKT Photonics (1 mentions)
Prior proscan 3, by Prior Scientific (1 mentions)
Axiovert 100, by Zeiss (1 mentions)
34 protocols using eclipse te2000 e inverted microscope
1
Quantifying HIV-1 Virus-Like Particle Formation
2
Assessing PD-L1 Expression in Pancreatic Cancer
3
Imaging Conjugate Biodistribution in U87ΔEGFR Glioma
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Intracranial U87ΔEGFR-luc tumor-bearing NSG mice (6–8 weeks old, male and female) were prepared as described above and randomized into three groups (n = 3) 5 days post tumor implantation. Each Cy5.5 conjugate was administered intravenously at 3 mg/kg. After 48 h, the tumor-bearing mice were anesthetized with ketamine/xylazine. Subsequently, the mice underwent cardiac perfusion with PBS(+) containing sodium heparin (10 units/mL) and then 4% paraformaldehyde/PBS(+). This step removes conjugates circulating or bound to the vascular endothelial cells. Major organs including the brain were then harvested. Cy5.5-based near-infrared fluorescence images of the harvested organs were taken using a LI-COR Odyssey 9120 imager (Ex: 685 nm laser, intensity: L1.0 for brain, L2.0 for other organs, Em: 700 nm channel). Semi-quantification of the signals from ROIs was also performed using LI-COR Image Studio software. For tissue imaging, the brain samples were embedded in paraffin and tissue sections were prepared (thickness: 5 μm). After de-paraffinization of using toluene, mounting medium containing DAPI (VECTOR #H-1200) was applied to the tissue slides. Fluorescence Images were taken using a Nikon Eclipse TE2000E inverted microscope (Cy5 channel). Three ROIs in each sample were acquired and analyzed for semi-quantification using ImageJ software.
4
Imaging Conjugate Biodistribution in U87ΔEGFR Glioma
5
Sulforaphane Inhibits Tumor Growth in Nude Mice
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The use and care of animals as models were approved by the Institutional Animal Care and Use Committee of Zaozhuang City Hospital (Zaozhuang, China). All experimental procedures were performed according to the approved guidelines. The mouse experiments were performed in the Animal Laboratory Center at Zaozhuang City Hospital. The cells (1×107cells) treated with various concentrations of sulforaphane, described above, were suspended in 100 μl serum-free medium and injected subcutaneously into the left flank of 4–6-week-old male BALB/c nu/nu nude mice (15–18 g), with 8 mice in each groups, 32 mice in total, were injected with cells. The mice were raised in the SPF animal facility with sterile food and water ad libitum, at 20°C with a 50% humidity and a 12-h light/dark cycle. Tumor size was measured with digital calipers and calculated every week. Tumor volume was measured every 7 days and at the end (~6 weeks) of treatment, when the mice were sacrificed. The tumors were excised, weighed, fixed in 10% neutral formalin, and embedded in paraffin for histological analysis under an Eclipse TE2000E inverted microscope (Nikon Corporation).
6
Characterization of Silicon Micropillars
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The morphology and structure of the macroporous silicon and subsequent silicon dioxide micropillars were characterized by scanning electron microscopy (SEM) using a FEI Quanta 600 environmental scanning electron microscope (FEI, Hillsboro, OR, USA) operating at an accelerating voltage between 15 and 25 kV. The micropillars were also morphologically characterized by transmission electron microscopy (TEM) using a JEOL 1011 (JEOL Ltd., Akishima-shi, Japan) operating in dark-field mode at 80 kV. Confocal laser scanning microscopy images were taken using a Nikon Eclipse TE2000-E inverted microscope, equipped with a C1 laser confocal system (EZ-C1 software, Nikon, Tokyo, Japan). A 488-nm helium-neon laser was used as excitation source for DOX-loaded micropillars. The emission was collected through a 590 ± 30 bandpass emission filter (red channel). All fluorescence images were captured using a 5-megapixel CCD. The concentrations of DOX were determined using a spectrofluorometer (PTI Quantamaster 40, Photon Technologies International, Edison, NJ, USA) at an exciting wavelength of 480 nm.
7
Necramed Treatment in Nude Mice
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Control (gavage, daily, 50% PEG400) or Necramed (chloroquine (30 mg/kg), nelfinavir (250 mg/kg), rapamycin (2.24 mg/kg), metformin (150 mg/kg), and dasatinib (4 mg/kg) in 50% PEG400, gavage, daily) treatment began at 10 weeks of age. For histology, nude mice were treated for 7 days and then euthanized 3 hours following the last treatment. Liver, heart, and kidneys were harvested, fixed in 10% buffered formalin for 24 hours and then 70% ethanol for at least 24 hours, blocked, sectioned with 3 μm width, and H&E stained by the core facility at the Moores Cancer Center. Histology images were taken with a 4X objective on a Nikon Eclipse TE2000E inverted microscope. An additional 3 pairs of C57BL/6 mice were treated with control or Necramed for 21 days, sacrificed, and blood collected immediately by cardiac extraction. Blood was centrifuged in a Micro Serum Separator (Professional Hospital Supply) and plasma saved in a microcentrifuge tube at 20°C until analysis by a VetScan instrument using a VS Complete Diagnostic Profile.
8
Live-cell imaging of cell migration
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Twenty-four hours after transfection, cells were either plated on fibronectin (10 μg/ml)-coated, 35-mm glass-bottom imaging dishes (MatTek, Ashland, MA) or FluoroDish cell culture dishes (35 or 50 mm; World Precision Instruments, Sarasota, FL) or were embedded in collagen gels as described and imaged the next day. Before imaging, growth medium was replaced with CO2-independent imaging medium containing 134 mM NaCl, 5.4 mM KCl, 1.0 mM MgSO4, 1.8 mM CaCl2, 20 mM HEPES, and 5 mM d -glucose (pH 7.4). A stage warmer (NevTek Airstream, Williamsville, VA) was used to maintain cells at 37°C, and cells were imaged on an Eclipse TE-2000E inverted microscope (Nikon) equipped with a 40×/1.3 numerical aperture (NA) Plan Fluor or 60×/1.4 NA Plan Apo oil-immersion objective, the appropriate fluorophore-specific filters (Chroma Technology, Bellows Falls, VT), a Spectra X LED light engine (Lumencor, Beaverton, OR), and a Clara charge-coupled device camera (Andor, Concord, MA). Images were acquired every 5–60 s with 500- to 800-ms exposure times.
9
Quantitative Analysis of Ras Surface Density and Mobility Using Dual-Colour FCS
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Surface density and mobility of Ras were quantitatively analysed using FCS. Dual-colour FCS was performed on a home-built setup based on a Nikon Eclipse TE2000-E inverted microscope46 (link). Briefly, excitation wavelengths were selected by bandpass filters from a pulsed white light laser source (SuperK Extreme EXW-12, NKT Photonics) and combined through a single-mode optical fibre. The excitation pulses enter the microscope via a multi-colour dichroic cube (Di01-R405/488/561/635-25x36, Semrock). The fluorescence signal is collected by a × 100 high-numerical aperture oil-immersion objective and recorded by avalanche photodiode detectors (Hamamatsu). The signal is directly converted into autocorrelation signal by a hardware correlator (Correlator.com). Lights (488 and 568 nm) were used to excite the Atto 488 fluorophore and Texas Red-DHPE, respectively. The resulting autocorrelation G(τ) was fit to the two-dimensional Gaussian diffusion model to calculate surface density and mobility of Ras46 (link).
10
Motility Analysis of Tagged RSP6 Strains
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