Te2000

Manufactured by Nikon

Sourced in Japan, United States, China, France, Germany, United Kingdom, Canada

The TE2000 is a compact and versatile inverted microscope designed for a wide range of laboratory applications. It features a high-quality optical system, a stable and user-friendly design, and a range of accessories to support various imaging techniques. The TE2000 is a reliable and efficient tool for researchers and scientists working in various fields, including cell biology, tissue engineering, and materials science.

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Lab products found in correlation

Transwell chamber, by Corning (9 mentions) Matrigel, by BD (7 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (7 mentions) Ez c1, by Nikon (6 mentions) Lsm 710 confocal microscope, by Zeiss (6 mentions) Ds qi1mc, by Nikon (5 mentions) Matrigel, by Yeasen (5 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Nis element br 3, by Nikon (4 mentions) Trypsin, by Thermo Fisher Scientific (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (4 mentions) Metamorph software, by Molecular Devices (4 mentions) Lab tek chambered coverglass slides, by Thermo Fisher Scientific (4 mentions) Lsm 780, by Zeiss (4 mentions) Gfp lc3 vector, by Genechem (3 mentions) Fura 2 am, by Thermo Fisher Scientific (3 mentions) Tecnai g2 12, by Thermo Fisher Scientific (3 mentions) C2 plus, by Nikon (3 mentions) Ds fi1, by Nikon (3 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (3 mentions)

456 protocols using te2000

Quantifying Renal and Cell ROS Levels

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The levels of ROS from renal tissues were assessed using the dihydroethidium (DHE) fluorescent probe (D7008, Sigma-Aldrich, MO, United States), following the previously described protocols (Fan et al., 2021 (link)). In brief, after frozen sections were incubated with 50 uM DHE for 1 h in the dark at room temperature, the sections were incubated for 10 min with DAPI (1 mg/ml). The cell images were taken with a fluorescent microscope (TE-2000, Nikon, Co., Japan) with an excitation (Ex) wavelength at 525 nm and an emission (Em) wavelength at 610 nm after washing. The values used ImageJ software to capture the red (the oxidized probe) and blue (the reduced probe) fluorescence intensities to calculate the ratio and then standardized to the con group.
The levels of ROS in HK-2 cells were tested by the 2′, 7′- dihydrofluorescein diacetate (DCFH-DA) fluorescent probe (D6883, Sigma-Aldrich, United States), following the manufacturer’s protocols. Briefly, after treating HK-2 cells with 10 uM DCFH-DA in the dark for 1 h, the pictures were detected using a fluorescent microscope (TE-2000, Nikon, Co., Tokyo, Japan) at (Ex/Em) 485 nm/530 nm.

Qiongyue Z., Xin Y., Meng P., Sulin M., Yanlin W., Xinyi L, & Xuemin S. (2022). Post-treatment With Irisin Attenuates Acute Kidney Injury in Sepsis Mice Through Anti-Ferroptosis via the SIRT1/Nrf2 Pathway. Frontiers in Pharmacology, 13, 857067.

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Assessing Oxidative Stress in Lung Cells

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The levels of ROS in lung tissues were assessed using the dihydroethidium (DHE) fluorescent probe (D7008, Sigma-Aldrich, MO, USA) following the previously described protocols [30 ]. In brief, frozen sections were stained with 50 uM of DHE for 1 hour at room temperature in the dark and then with DAPI (1 mg/ml) for 10 minutes. The fluorescence intensity was measured using a fluorescent microscope (TE-2000, Nikon Co., Tokyo, Japan) at 525 nm excitation (Ex) wavelength and 610 nm emission (Em) wavelength after washing.
ROS levels in MLE-12 cells were tested by the 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe (D6883, Sigma-Aldrich, MO, USA) following the manufacturer's protocols. After treating MLE-12 cells with 10 uM DCFH-DA for 1 hour in the dark and then treated with DAPI (1 ug/ml) for 10 minutes, the cell pictures were detected using a fluorescent microscope (TE-2000, Nikon, Co., Tokyo, Japan) at (Ex/Em) 485 nm and 530 nm.

Wang Y., Dong Z., Zhang Z., Wang Y., Yang K, & Li X. (2022). Postconditioning with Irisin Attenuates Lung Ischemia/Reperfusion Injury by Suppressing Ferroptosis via Induction of the Nrf2/HO-1 Signal Axis. Oxidative Medicine and Cellular Longevity, 2022, 9911167.

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Fluorescence Microscopy Imaging Techniques

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Stained sections were investigated with an inverted fluorescence microscope (Nikon TE2000; Nikon, Japan) equipped with a spot digital camera with three filters (for emission spectra maxima at 358, 461 and 555 nm). Both microscope and camera are connected to a computer system installed with image software (NIS Element BR-3.2; Nikon) including image merging and a fluorescence intensity analyzer. For laser confocal microscopy, the same microscope was used, which is equipped with a laser emission and detection system with three different channels. The optical scanning and image-processing tasks were run by the program Nikon EZ-C1 (v.3.80) including reconstruction of Z-stack images into projections or 3D images.

Liu W., Atturo F., Aldaya R., Santi P., Cureoglu S., Obwegeser S., Glueckert R., Pfaller K., Schrott-Fischer A, & Rask-Andersen H. (2015). Macromolecular organization and fine structure of the human basilar membrane - RELEVANCE for cochlear implantation. Cell and Tissue Research, 360(2), 245-262.

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Transwell Assay for HUVEC Invasion

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HUVECs invasion activities were determined using a 24-well Transwell chamber (cat. no. 3577; Corning Costar Corp.) with Matrigel (cat. no. 356234; BD Biosciences). Cells (1×10⁵) prepared in 400 µl serum-free media were seeded into the upper chamber of the Transwell insert. A total of 600 µl TCM was added to the lower chamber of the Transwell insert. After incubation for 48 h, the cells remaining in the upper chamber of the Transwell insert were removed. Cells on the underside of the chamber were fixed with 4% paraformaldehyde at room temperature for 30 min and stained with 0.1% crystal violet at room temperature for 20 min. Invaded cells were subsequently counted under a inverted fluorescence microscope (Nikon TE2000; Nikon Corp.) (magnification, ×200) in six different fields of view for each sample.

Guo W., Li H., Liu H., Ma X., Yang S, & Wang Z. (2019). DEPDC1 drives hepatocellular carcinoma cell proliferation, invasion and angiogenesis by regulating the CCL20/CCR6 signaling pathway. Oncology Reports, 42(3), 1075-1089.

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Colony Formation Assay Protocol

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After treatment, cells were seeded into 6-well plates (400 cells/well) and further cultured at 37°C. After incubation for 14 days, the cells were washed with PBS, fixed with 4% paraformaldehyde at room temperature for 15 min, and stained with 0.2% crystal violet solution at room temperature for 20 min. The number of colonies was counted with an inverted fluorescence microscope (Nikon TE2000; Nikon Corp.).

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Endothelial Cell Migration Assay

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Migration of HUVEC cells was monitored by an artificial scratch in a monolayer of confluent endothelial cells with a 1000 µl tip 24 h after tranfection and observation of the extent of wound closure after 24 or 48 h. Then the sharp wound in each well was captured using an inverted microscope (Nikon TE 2000; Nikon).

Duan Q., Yang L., Gong W., chaugai S., Wang F., Chen C., Wang P., Zou M, & Wang D.W. (2015). MicroRNA‐214 Is Upregulated in Heart Failure Patients and Suppresses XBP1‐Mediated Endothelial Cells Angiogenesis. Journal of Cellular Physiology, 230(8), 1964-1973.

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Autophagic Flux Measurement via AO Staining

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The degradation of autophagosomes is mediated by lysosomes; autolysis lowers the pH value and allows AO dye to penetrate into the acidic organelles and exhibit a red fluorescence; therefore, the intensity of red fluorescence following AO staining can be used to indicate the levels of autolysosomes (16 (link)). Cells were treated with dimethyl sulfoxide (DMSO) or 40 µM wogonin for 24 h in DMEM supplemented with 10% FBS at 37°C in an atmosphere containing 5% CO₂, then washed three times with PBS, fixed with 4% paraformaldehyde for 15 min, stained with 1 mg/l AO dye solution for 10 min at 37°C and then imaged under a fluorescence microscope (Nikon TE2000; Nikon Corporation, Tokyo, Japan).

Li S.J., Sun S.J., Gao J, & Sun F.B. (2016). Wogonin induces Beclin-1/PI3K and reactive oxygen species-mediated autophagy in human pancreatic cancer cells. Oncology Letters, 12(6), 5059-5067.

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Cholesterol Levels and Absorption in Chondrocytes

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TC levels in synovial fluid, serum, cells and cartilages were detected using a Total Cholesterol Assay Kit (Solarbio, Beijing, China) according to the manufacturer's instruction and the detailed method was shown in Additional file 1. Nitrobenzoxadiaz (NBD)‐cholesterol (Biofound Biotechnology Co, Beijing, China), a fluorescent cholesterol analogue, was used to assess the cholesterol absorption level of cells. Briefly, chondrocytes were cultured for 24 h in serum‐free DMEM containing 5 μg/mL NBD‐cholesterol with or without 10 μg/mL IL‐1β (Peprotech, NJ, USA), 50 μg/ml Tumor necrosis factor α (TNFα, Peprotech), or hydrogen peroxide. After the treatment, the cells were fixed in 4% paraformaldehyde (PFA) for 15 min and stained with DAPI (Sigma–Aldrich) for 20 min. The cells were observed using a fluorescence microscope (NIKON TE2000, Nikon Corporation, Japan).

Li J., Li X., Zhou S., Wang Y., Ying T., Wang Q., Wu Y, & Zhao F. (2023). Circular RNA circARPC1B functions as a stabilisation enhancer of Vimentin to prevent high cholesterol‐induced articular cartilage degeneration. Clinical and Translational Medicine, 13(9), e1415.

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Fluorescent Staining of Actin and Nuclei

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Cells were incubated for 24 h at 37°C with 5% CO₂, in a humidified incubator, then fixed in 3.7% formaldehyde for 20 min at room temperature, and stained for actin cytoskeleton and nuclei. Cells were permeabilized with 0.5% Triton X-100 in PBS at 4°C for 15 min, incubated for 1 h with TRITC-labeled phalloidin (1:200) at 37°C in the dark, then with Hoechst 33342 (Life Technologies, Carlsbad, CA, USA) for 10 min at room temperature, and washed twice with deionized water. Samples were observed under fluorescence microscopy (Nikon TE2000, Nikon Instruments, Amsterdam, Netherlands) at an excitation wavelength of 360 nm for Hoechst staining (nucleus staining) and 630 nm for phalloidin (actin staining).

Collart-Dutilleul P.Y., Panayotov I., Secret E., Cunin F., Gergely C., Cuisinier F, & Martin M. (2014). Initial stem cell adhesion on porous silicon surface: molecular architecture of actin cytoskeleton and filopodial growth. Nanoscale Research Letters, 9(1), 564.

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Quantifying Cellular and Mitochondrial ROS

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The ROS levels were measured using a dihydroethidium (DHE) fluorescent probe for cytosolic ROS detection or MitoSOX Red (ThermoFisher Scientific, Waltham, MA) for mitochondrial ROS detection. DHE and MitoSOX staining and detection were conducted as previously described.26 The cellular H₂O₂ levels were detected using a Hydrogen Peroxide Assay Kit (Beyotime, Shanghai, China). Tissue sections were frozen in optimal cutting temperature compound and quickly sectioned, and the sections were then stained with DHE or MitoSOX dye according to the manufacturers' protocols. To quantify the intensity of DHE and MitoSOX fluorescence, the plates were measured using a Fluoroskan Ascent Fluorometer (ThermoFisher, Helsinki, Finland). The ROS concentrations are expressed as a normalized value relative to the fluorescence intensity of the control cells. To visualize the staining, the sections or specimens were placed on an inverted fluorescence microscope (Nikon TE2000, Nikon Corporation, Tokyo, Japan).

Wang B., Xiong S., Lin S., Xia W., Li Q., Zhao Z., Wei X., Lu Z., Wei X., Gao P., Liu D, & Zhu Z. (2017). Enhanced Mitochondrial Transient Receptor Potential Channel, Canonical Type 3–Mediated Calcium Handling in the Vasculature From Hypertensive Rats. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(7), e005812.

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