Eclipse ti s inverted fluorescence microscope

Manufactured by Nikon

Sourced in Japan, United States

The Eclipse Ti-S is an inverted fluorescence microscope designed for advanced imaging applications. It features a high-resolution optical system, a motorized stage, and supports a range of fluorescence imaging techniques. The Eclipse Ti-S is a versatile tool for researchers and scientists working in fields such as cell biology, neuroscience, and material science.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Uv1050, by Techcomp (2 mentions) Lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions) Fitc cona, by Merck Group (1 mentions) Antifade mounting medium, by Beyotime (1 mentions) Paraformaldehyde (pfa), by Leagene (1 mentions) Nis elements br 3, by Nikon (1 mentions) Flow cytometry, by BD (1 mentions) Agilent 7500ce icp ms, by Agilent Technologies (1 mentions) Nicolet is5 instrument, by Thermo Fisher Scientific (1 mentions) Hplc system, by Agilent Technologies (1 mentions) Eclipse ci, by Nikon (1 mentions) Su8020, by Hitachi (1 mentions) Thermo forma 311, by Thermo Fisher Scientific (1 mentions) C6 plus, by BD (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Beyotime (1 mentions) Nanodrop 2000 ultra micro spectrophotometer, by Thermo Fisher Scientific (1 mentions) Nicolet is5, by Thermo Fisher Scientific (1 mentions) Digital sight ds u2 camera, by Nikon (1 mentions)

29 protocols using eclipse ti s inverted fluorescence microscope

Time-lapse Fluorescence Microscopy of Bacterial Biofilm

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Strains N°24 and N°40 were incubated in a 4-well glass Lab-Tek®II Chamber Slide System (NUNC, Denmark) while inclined at approximately 45° to form a clear liquid-air interface, and placed in a moist sterile incubation box. The FLM assay was performed in the same cultures after 12, 24, 36, 48, 60, 72, 84, 96, 108, and 120 h incubation period. Treatments for each strain were in triplicates. The wells were rinsed with PBS and fixed with 4% paraformaldehyde (LEAGENE, China) for 30 min. Then, the wells were labeled by FITC-ConA (Sigma, USA) for exopolysaccharides (EPS) and propidium iodide (PI; Sigma, USA) for bacterial nucleic acid, before rinsed finally with PBS. The slide was sealed with antifade mounting medium (Beyotime, China).
The pictures were taken with a Nikon ECLIPSE Ti-S Inverted Fluorescence Microscope (Nikon, Japan) equipped with 33 mm ND4/ND8 filters employing green filter detecting PI fluorescence (500–550/615 nm excitation/emission wavelengths) and blue filter to detect FITC fluorescence (400–490/525 nm excitation/emission wavelengths). The pictures were processed with NIS-Elements BR 3.0 software (Nikon, Japan).

Liu J., Fu K., Wang Y., Wu C., Li F., Shi L., Ge Y, & Zhou L. (2017). Detection of Diverse N-Acyl-Homoserine Lactones in Vibrio alginolyticus and Regulation of Biofilm Formation by N-(3-Oxodecanoyl) Homoserine Lactone In vitro. Frontiers in Microbiology, 8, 1097.

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Measuring Intracellular ROS in hAMSCs and BMSCs

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The intracellular ROS level for hAMSCs and BMSCs was measured using the ROS detection kit from Applygen (red fluorescence) and Beyotime (green fluorescence), respectively. The cells were washed with D‐PBS after incubation with dihydroethidium for 30 min at 24–28°C. Finally, red and green fluorescent protein expression was measured using Nikon Eclipse Ti‐S inverted fluorescence microscope (Nikon) and flow cytometry (BD Biosciences).

Yuan H., Xu Y., Luo Y., Zhang J., Zhu X, & Xiao J. (2022). Ganoderic acid D prevents oxidative stress‐induced senescence by targeting 14‐3‐3ε to activate CaM/CaMKII/NRF2 signaling pathway in mesenchymal stem cells. Aging Cell, 21(9), e13686.

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Immunofluorescence Detection of P-glycoprotein

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Cells were seeded onto the cover slips and incubated for 24 h to attach. Cells were fixed and permeabilized with methanol for 10 min at −20 °C. Cells were washed three times, five minutes each in PBS, and then blocked with 1% BSA in PBST for 1 h at room temperature. Later, cells were incubated with mouse monoclonal anti-P-gp antibody diluted 1:100 (Santa Cruz, USA) in 1% BSA in PBST in a humidified chamber for 1 h at room temperature. Subsequently, cells were washed five times, five minutes each in PBS and incubated with anti-mouse IgG secondary antibody conjugated with FITC (1:100) (Santa Cruz, USA), for 1 h at room temperature in the dark. Finally, cells were washed five times, five minutes each in PBS and mounted on a glass slide using 90% glycerol. Images were acquired using a Nikon Eclipse Ti-S inverted Fluorescence microscope (Nikon Instruments, Melville, New York).

Syed S.B., Arya H., Fu I.H., Yeh T.K., Periyasamy L., Hsieh H.P, & Coumar M.S. (2017). Targeting P-glycoprotein: Investigation of piperine analogs for overcoming drug resistance in cancer. Scientific Reports, 7, 7972.

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Evaluation of Cu-DCA Nanoparticles Against Oxidative Stress

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Briefly, HUVECs were seeded onto 6-well plates with a density of 5 × 10⁴ cells per well. After 24 h incubation, except of the control group and Cu-DCA NZs group, H₂O₂ (150 µM) was added to the wells. Subsequently, the samples (PBS, DCA and Cu-DCA NZs solution) were immediately added for incubation of 24 h. Then, the group of Cu-DCA NZs + H₂O₂ + NIR wells was exposed to laser irradiation (808 nm, 1 W·cm^− 2, 5 min). After irradiation, the DCFH-DA probe was added according to the instructions of the reactive oxygen detection kit and incubated for 30 min. Finally, the fluorescence images were taken by a Nikon Eclipse Ti-S inverted fluorescence microscope (Nikon Corporation, Japan).

Huang W., Xu P., Fu X., Yang J., Jing W., Cai Y., Zhou Y., Tao R, & Yang Z. (2023). Functional molecule-mediated assembled copper nanozymes for diabetic wound healing. Journal of Nanobiotechnology, 21, 294.

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Comprehensive Characterization of Nanomaterials

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The following instruments were used: an ultrasonic cleaner (42 kHz, 100 W, Kun Shan Ultrasonic Instruments), an AFM (SHIMADZU), an HPLC system (Agilent Technologies), a multifunctional microplate inspection system (Thermo Fisher), a DLS particle size analyser (Malvern), a UV–vis absorption spectrophotometer (UV1050, Techcomp). a Nicolet iS5 instrument (Thermo Fisher Scientific, USA) for Fourier transform infrared (FT-IR) spectroscopy, an InVivo Elite Preclinical Optical Imaging System (InVivo Smart, VIEWORKS), a field emission TEM (Thermo Fisher), an SU8020 (HITACHI, Japan) for field emission SEM, an Agilent 7500ce ICP–MS (USA), a Nikon Eclipse Ti-S inverted fluorescence microscope (Nikon Corporation, Japan), an upright microscope (Nikon ECLIPSE Ci, Nikon Corporation), a fully automatic chemiluminescence fluorescence image analysis system (Tanon 5200, Shanghai Tanon), and a portable dissolved oxygen analyser (JPBJ-608, Shanghai INESA).

Fu X., Yu X., Jiang J., Yang J., Chen L., Yang Z, & Yu C. (2022). Small molecule-assisted assembly of multifunctional ceria nanozymes for synergistic treatment of atherosclerosis. Nature Communications, 13, 6528.

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Cytotoxicity Evaluation of PC-DOX in CT26 Cells

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CT26 (mouse colon cancer cells) was cultured in DMEM containing 10% (v:v) FBS (Gibco Life Technologies, USA) and 1% (v:v) penicillin–streptomycin (Beyotime Biotechnology, Shanghai, China). Cells were incubated at 37 °C in a humidified incubator with 5% CO₂ (Thermo Forma 311, Thermo Scientific, Waltham, MA, USA).
CT26 cells in the logarithmic growth phase were rinsed with PBS, digested with trypsin, then centrifuged at 1500 rpm for 3 min to pellet the cells. The cells were then counted using a cell counter, and seeded into a 24-well plate at a density of 15 × 105 cells·well-1. At the end, the cells were cultured for about 16–18 h.
The preparation and enzymolysis of PC–DOX could refer to the steps in Section 4.4. After enzymolysis, PC–DOX was diluted with DMEM (DOX, 2.5 μg/mL), 1 mL of diluted sample was incubated with the cells for 4 h and cultured at 37 °C in a 5% CO₂ incubator. After that, the cells were washed with PBS and imaged using a Nikon eclipse Ti-S inverted fluorescence microscope (Ti-S, Nikon Corporation, Tokyo, Japan).
Finally, the cells were digested with trypsin, then centrifuged at 1500 rpm for 3 min to pellet the cells. Using the DMEM medium to resuspend the cells and using a flow cytometer (BD C6plus, Waltham, MA, USA), we quantified the fluorescence intensity in cells.

Li J., Ma L., Wang C., Jiang P., Cui P, & Wang J. (2022). Rationally Designed Oral DOX Gels for Colon-Specific Administration. Gels, 8(12), 759.

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Multimodal Characterization of Nanomaterials

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The following instruments were used: a Bruker Avance III 600 MHz nuclear magnetic resonance (NMR) spectrometer (Bruker, Germany); a dynamic light scattering particle size analyzer (DLS, Malvern, UK), an ultraviolet‒visible (UV–vis) absorption spectrophotometer (UV1050, Techcomp, China); a Nicolet iS5 (Thermo Fisher Scientific, USA) for Fourier transform infrared (FTIR) spectroscopy; a Nanodrop2000 ultramicrospectrophotometer (Thermo Fisher Scientific, USA); an LC20 gel permeation chromatography (GPC, Shimadzu, Japan); a Nikon Eclipse Ti‐S inverted fluorescence microscope (Nikon Corporation, Japan); a field emission TEM (FEI, Netherlands); an NCF950 laser scanning confocal microscope (LSCM, Yongxin, China); an Illumina HiSeq2500 high throughput sequencer (Illumina, USA); and a flow cytometer (TCM, Beckman Coulter, USA).

Zhao Y., Li Y., Wang F., Gan X., Zheng T., Chen M., Wei L., Chen J, & Yu C. (2023). CES1‐Triggered Liver‐Specific Cargo Release of CRISPR/Cas9 Elements by Cationic Triadic Copolymeric Nanoparticles Targeting Gene Editing of PCSK9 for Hyperlipidemia Amelioration. Advanced Science, 10(19), 2300502.

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Comparative Adipose Tissue Model

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Microscopic imaging was done with a Nikon Eclipse Ti-s inverted fluorescence microscope (Nikon, Tokyo, Japan) and Nikon digital sight DS-U2-camera (Nikon), and images were further processed with NIS Elements (Nikon), ZEN 2012 software (Carl Zeiss, Oberkochen, Germany). Confocal imaging was done with LSM710 and with Zeiss Axio Observer Z1 inverted microscope culture and the vascularized adipose tissue model contained lipid accumulating cells, which the undifferentiated cells did not contain (Fig. 1). Upon morphological inspection, the lipid accumulation in the vascularized adipose tissue model was more evenly spread out than in adipocyte culture, where there were fewer adipocytes that contained larger lipid storages. This morphology was also described in our previous study (Huttala et al., 2018) (link) but could not be confirmed by the quantitative results of lipid per cell measurements (Fig. 1). Of the secreted proteins studied, adiponectin was significantly increased in the vascularized adipose tissue model (Fig. 1), but the secretion of leptin and FABP4 was similar in both models.

Huttala O., Sarkanen J.R., Heinonen T, & Ylikomi T. (2019). Presence of vasculature results in faster insulin response in adipocytes in vascularized adipose tissue model. ALTEX, 36(3).

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Oleocanthal Modulates ERα Expression

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To evaluate the effect of (−)-oleocanthal on expression and localization of ERα, BT-474 cells were seeded on 8-chamber culture slides (Becton Dickinson and Company, NJ, USA) at 4×10⁴ cells/chamber (2 replicates/group) and allowed to attach overnight in medium supplemented with 10% FBS. Cells were then washed in PBS and incubated with vehicle control or treatment medium containing various concentrations of (−)-oleocanthal in mitogen-free or 10 nM 17β-estradiol-supplemented media for 24 h. At the end of treatment, cells were washed in pre-cooled PBS, and fixed with 1:1 volume of methanol: acetone precooled to − 20°C for 2 min. Fixed cells were washed in PBS and blocked with 2% BSA-TBST for 1 h at room temperature. Afterwards, BT-474 cells were incubated in primary antibody to ERα (1:3000) in 2% BSA-TBST overnight at 4°C. Thereafter, cells were washed in pre-cooled PBS followed by incubation with goat anti-rabbit Alexa Fluor 594-conjugated secondary antibody (1:3000) in 2% BSA-TBST for 1 h at room temperature. After final washing, cells were embedded in Vectashield mounting medium with DAPI (Vector Laboratories IN, Burlingame, CA, USA). Fluorescent images were captured using Nikon Eclipse Ti–S inverted fluorescence microscope (Norcross, GA, USA) at a magnification of 200×.

Ayoub N.M., Siddique A.B., Ebrahim H.Y., Mohyeldin M.M, & El Sayed K.A. (2017). The olive oil phenolic (−)-oleocanthal modulates estrogen receptor expression in luminal breast cancer in vitro and in vivo and synergizes with tamoxifen treatment. European journal of pharmacology, 810, 100-111.

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Visualizing Chromatin Condensation in MN9D Cells

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MN9D cells grown in glass-bottomed plates were treated with 250 μM MPP⁺ or 4'I- MPP⁺ for 12 h and then with 300 nM DAPI in KRB-HEPES for 15 min in the dark at 37 °C [24 (link)]. Then cells were rinsed and PBS was added to cover the cell layer. Increase in nuclear DAPI fluorescence (Ex/Em at 358/461 nm) due to chromatin condensation was observed using a Nikon ECLIPSE Ti-S inverted fluorescence microscope. The controls were treated similarly except that MPP⁺ or 4'I-MPP⁺ was omitted from the initial incubation medium.

Mapa M.S., Le V.Q, & Wimalasena K. (2018). Characteristics of the mitochondrial and cellular uptake of MPP+, as probed by the fluorescent mimic, 4'I-MPP+. PLoS ONE, 13(8), e0197946.

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