Ti u inverted fluorescence microscope

Manufactured by Nikon

Sourced in Japan, United States

The Ti-U inverted fluorescence microscope is a high-performance laboratory instrument designed for advanced imaging applications. It features a stable, inverted optical configuration and supports a range of fluorescence techniques. The Ti-U provides researchers with a versatile platform for their imaging needs.

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Lab products found in correlation

Basic fibroblast growth factor (bfgf), by Merck Group (1 mentions) Ultra low attachment plate, by Corning (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) B27 supplement, by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions) Nis element software, by Nikon (1 mentions) Matrigel, by BD (1 mentions) Pierce anti ha magnetic beads, by Thermo Fisher Scientific (1 mentions) Easysep magnet, by STEMCELL (1 mentions) C4742 95, by Nikon (1 mentions) Ti u inverted microscope, by Nikon (1 mentions) Gfp quantification kit, by Abcam (1 mentions) Victor x4, by PerkinElmer (1 mentions) Entinostat, by Cayman Chemical (1 mentions) Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (1 mentions) Celltracker blue cmac dye, by Thermo Fisher Scientific (1 mentions) Celltracker green cmfda, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Eclipse Ti-U inverted fluorescence microscope Ti-U inverted microscope Ti-U fluorescence microscope Nikon Ti inverted fluorescence microscope Ti-U microscope Ti inverted microscope Ti fluorescence microscope Inverted fluorescence microscope Ti inverted Ti microscope

15 protocols using ti u inverted fluorescence microscope

Detecting pSV-Tag-MP Expression in Tumor Cells

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For checking the expression and location of pSV-Tag-MP in tumor cells, 2 × 10⁵ Hey cells were transfected with either pSV-GFP or pSV-Tag-MP in 12-well plates. The cells were imaged 48 h after transfection under the Nikon Ti-U Inverted Fluorescence Microscope.
For the cell pull-down experiment, 5 × 10⁶ PBMCs were spiked with 100 Hey cells that had been incubated with 2 IU of either pSV-GFP or pSV-Tag-MP for 72 h. Cells were incubated in 2-mL tubes with 50 μL of Pierce Anti-HA Magnetic Beads (Thermo Fisher Scientific, Waltham, MA, USA) in 1 mL of 0.1% BSA-PBS by rotation for 30 min at 4°C. Afterward, the tubes were loaded to the EasySep Magnet (STEMCELL Technologies, Cambridge, MA, USA) for 1 min to allow the tagged tumor cells to attach to the magnetic side of the tube. PBS with the unbound cells was discarded. After two washes with 1 mL of 0.1% BSA-PBS, the beads with bound cells were suspended in 100 μL of DMEM containing 10% FBS and seeded in a 96-well plate. The pull-down cells were analyzed under a Nikon Ti-U Inverted Fluorescence Microscope.

Fu X., Tao L, & Zhang X. (2021). A chimeric virus-based probe unambiguously detects live circulating tumor cells with high specificity and sensitivity. Molecular Therapy. Methods & Clinical Development, 23, 78-86.

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Mammosphere Assay for Cancer Stem Cells

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Mammosphere assay was performed as previously described (Lo et al., 2012 (link)). Material and reagent were prepared as followed; a 96-well ultra low attachment plate (Corning) and DMEM/F12 media containing 20 ng/ml epidermal growth factor (Sigma Aldrich), 10 ng/ml basic fibroblast growth factor (Sigma Aldrich), 5 μg/ml insulin (Sigma Aldrich), 50 × B27 supplement (Life technologies) and 0.4% FBS (Gibco). Cells were counted and seeded 200 cells (in 200 μl) per well in the ultra low attachment plate with mixed mammosphere media. For treatment, 10 replicates were seed and incubated with either DMSO or RORγ small molecules for 6 days. Images were taken using the Nikon Ti–U inverted fluorescence microscope under bright field.

Oh T.G., Wang S.C., Acharya B.R., Goode J.M., Graham J.D., Clarke C.L., Yap A.S, & Muscat G.E. (2016). The Nuclear Receptor, RORγ, Regulates Pathways Necessary for Breast Cancer Metastasis. EBioMedicine, 6, 59-72.

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Metastatic B16F10 Cell Trafficking via LPA Receptor

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In some experiments, 1 × 10⁶ B16F10 cells stably expressing GFP were injected via tail vein into the various LPA receptor KO female mice and their respective WT littermates. Mice were sacrificed 6 h and 24 h after injection. Lungs were harvested and filled with PBS, and the GFP-expressing B16F10 cells on the lung surface were imaged using a Nikon TiU inverted fluorescence microscope (using a low-magnitude 4X objective lens). At least 12 images were taken for each mouse, and tumor burden was measured as total area of GFP fluorescence (in pixel units) using the Nikon NIS element software.

Lee S.C., Fujiwara Y., Liu J., Yue J., Shimizu Y., Norman D.D., Wang Y., Tsukahara R., Szabo E., Patil R., Banerjee S., Miller D.D., Balazs L., Ghosh M.C., Waters C.M., Oravecz T, & Tigyi G.J. (2014). Autotaxin, LPA Receptors (1 and 5) Exert Disparate Functions in Tumor Cells Versus the Host Tissue Microenvironment in Melanoma Invasion and Metastasis. Molecular cancer research : MCR, 13(1), 174-185.

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Wound Healing Assay with RORγ Knockdown

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Wound healing assay was performed as previously described (Oh et al., 2014 (link)). Cells were transfected with control-siRNA or RORγ-siRNA for 24 h in a 6-well plate. Scratches were created across the monolayers of cell lines using a sterilized yellow tip. Images of wounds were taken at 0-, 24-, and 48-h time points using a 4 × objective of the Nikon Ti–U inverted fluorescence microscope under bright field. Cell migration (gap closure) of scratch wounds was measured using the ImageJ software.

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Measuring Algal Cell Diameters

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Photomicrographs of algal cells were taken using an inverted Nikon microscope (Nikon Ti-U Inverted Fluorescence Microscope) and the cell diameters estimated both manually and using IMARIS image analysis software.

Chioccioli M., Hankamer B, & Ross I.L. (2014). Flow Cytometry Pulse Width Data Enables Rapid and Sensitive Estimation of Biomass Dry Weight in the Microalgae Chlamydomonas reinhardtii and Chlorella vulgaris. PLoS ONE, 9(5), e97269.

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Angiogenesis Assay using HUVEC

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The day before the experiment, 20 μL and 1 mL tips and 48-well plates in a 4°C refrigerator was prepared. matrigel™ (50 μL; cat. no. 356230; BD Bedford, MA, USA) was 4 degrees frozen for 2 hours and placed in an ice bath when the matrigel was in a liquid state. The matrix gel was diluted 3 times with a DMEM, and 150 μL matrigel-medium suspension was added to each well of the 48-well plate; the whole process was carried out in an ice bath. The treated plate was placed in a 37°C incubator for about 30 minutes. Inoculation of cells and administration were done as follows: HUVEC were digested in a logarithmic growth phase, centrifugated, and adjusted the cell density to 2 ∗ 10⁴, 200 μL of medium per well. Except for the control group, the other groups were treated with various concentrations of YPFS (10, 30, and 50 mg/mL) for 24 hours and resuspended in a DMEM with 10% FBS with or without TSLP (100 ng/mL). After approximately 6–10 hours, the tube began to form, and the images were obtained using the Nikon Ti-U inverted fluorescence microscope.

Yuan Q., Yao F., Zhou L., Liang G., Song X., Jiang G., Zhou M, & Zhang L. (2019). Yu Ping Feng San Exert Anti-Angiogenesis Effects through the Inhibition of TSLP-STAT3 Signaling Pathways in Hepatocellular Carcinoma. Evidence-based Complementary and Alternative Medicine : eCAM, 2019, 1947156.

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Quantifying Cellular Mitotic Abnormalities

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Still images were acquired using a Hamamatsu Orca CCD camera attached to a Nikon Ti-U inverted fluorescence microscope at 20× magnification in air (or 60× magnification in oil for metaphase spreads). Phase contrast and fluorescence images were merged using ImageJ. For multinucleation experiments, cells with two or more nuclei were counted and recorded as the percentage of the total number of cells in each image. The number of images needed to accurately quantify an experimental condition was determined using a running average. Briefly, the total percentage of multinucleated cells was adjusted as each additional image was analyzed. When the percentage of multinucleation stabilized, no further images were analyzed. Similar methodology was used to quantify centrosome amplification and multipolar mitosis. For metaphase spreads, total chromosomes per cell were counted for 30–36 cells. Aneuploidy was defined as the number of chromosomes per cell normalized to the average number of chromosomes per cell in a population.
Timelapse movies were acquired using a Hamamatsu C4742-95 camera attached to a Nikon Ti-U inverted microscope and fitted with an environmental chamber held at 90% humidity, 37°C, and 5% CO₂. Images were acquired at 10× magnification every 3 min for a total of 8 hr, and stacked in ImageJ.

Simi A.K., Anlaş A.A., Stallings-Mann M., Zhang S., Hsia T., Cichon M., Radisky D.C, & Nelson C.M. (2018). A soft microenvironment protects from failure of midbody abscission and multinucleation downstream of the EMT-promoting transcription factor Snail. Cancer research, 78(9), 2277-2289.

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Visualizing PKCα Distribution in Myocytes

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Images for GFP and brightfield microscopy were collected from live human myocytes 1–2 d after gene transfer. To determine the PKCα cellular distribution in rat myocytes, paraformaldehyde-fixed myocytes isolated 18–20 wk after sham and PO surgery were immunostained with PKCα (Santa Cruz Biotechnology) and Texas Red–conjugated goat anti-mouse Abs, as described earlier (Hwang et al., 2012 (link); Kim et al., 2016 (link)). Images were collected with a Nikon Ti-U inverted fluorescence microscope and DS-Fi1 5 megapixel digital imaging using identical exposure times for each sample.

Ravichandran V.S., Patel H.J., Pagani F.D, & Westfall M.V. (2019). Cardiac contractile dysfunction and protein kinase C–mediated myofilament phosphorylation in disease and aging. The Journal of General Physiology, 151(9), 1070-1080.

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Morusin Modulates HUVEC Tube Formation

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Matrigel™ (50 μL; BD Discovery Labware, Bedford, MA, USA) was tiled on the bottom of 96-well plates at 37°C for 45 min. HUVECs, which were treated with various concentrations of morusin (1, 2, and 3 μg/mL) for 24 h and re-suspended in DMEM with 10% FBS with or without VEGF (10 ng/mL), were seeded at a density of 2×10⁴ cells/well in 50 μL of medium. After approximately 6–10 h, the tube began to form, and the images were obtained using the Nikon Ti-U inverted fluorescence microscope. Tube formation was defined by counting the number of branch points of the formed tubes,32 (link) and the total tube lengths in five randomly selected microscopic fields per well were quantified using Image-Pro Plus 6.0 software.33 (link)

Gao L., Wang L., Sun Z., Li H., Wang Q., Yi C, & Wang X. (2017). Morusin shows potent antitumor activity for human hepatocellular carcinoma in vitro and in vivo through apoptosis induction and angiogenesis inhibition. Drug Design, Development and Therapy, 11, 1789-1802.

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Assessing Aβ42-Induced Cytotoxicity in PC12 Cells

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The PC12 culture conditions were the same as for 24-well plates, and 5 × 10⁴ cells were added to each well. Radiation was applied to the Aβ42 modeling group or control group, as mentioned in Section 3.5. Irradiation followed by 20 hours of culture, the culture solution was carefully removed, and calcein solution with a concentration of 0.5 M was added to each well, followed by co-incubation for 30 min. Live cells were observed under a Nikon Ti-U inverted fluorescence microscope (Tokyo, Japan).

Wang L., Cheng Y., Wang W., Zhao J., Wang Y., Zhang X., Wang M., Shan T, & He M. (2023). Effects of Terahertz Radiation on the Aggregation of Alzheimer’s Aβ42 Peptide. International Journal of Molecular Sciences, 24(5), 5039.

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