Ti2 e inverted microscope

Manufactured by Nikon

Sourced in Japan, United States

The Ti2-E inverted microscope is a high-performance laboratory instrument designed for advanced imaging and analysis. It features a sturdy inverted frame, allowing for a wide range of applications. The Ti2-E offers excellent optical quality, precise control, and efficient data capture capabilities to meet the demands of modern research and clinical settings.

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Lab products found in correlation

Hoechst, by Thermo Fisher Scientific (2 mentions) Nis elements software, by Nikon (2 mentions) Nis element, by Nikon (2 mentions) Sp8 confocal microscope, by Leica (2 mentions) 96 well dish, by Ibidi (2 mentions) Ds qi2 monochrome cmos camera, by Nikon (2 mentions) 60 plan apocr λ, by Nikon (2 mentions) Ax r confocal microscopy system, by Nikon (2 mentions) Live dead baclight bacterial viability assay, by Thermo Fisher Scientific (2 mentions) 24 well plate, by Ibidi (2 mentions) Csu w1 spinning disk, by Yokogawa (1 mentions) Biostation im q, by Nikon (1 mentions) Plan apochromat, by Yokogawa (1 mentions) L3224, by Thermo Fisher Scientific (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Lu nf laser launch, by Nikon (1 mentions) 3d n storm super resolution microscope, by Nikon (1 mentions) Co2 independent media, by Thermo Fisher Scientific (1 mentions) Prime bsi scmos camera, by Teledyne (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Inverted microscope (910 citations) Ti2-E (108 citations) Ti2 microscope (102 citations) Ti2 inverted microscope (50 citations) Ti2-E microscope (50 citations) Eclipse Ti2-E inverted microscope (33 citations) Ti2-E inverted fluorescence microscope (5 citations) Inverted Research Microscope Eclipse Ti2-E (4 citations) Ti2-E motorized inverted microscope (2 citations) Eclipse Ti2‐E inverted microscope system (2 citations)

54 protocols using ti2 e inverted microscope

Live Tracking of HIV Integration in THP-1 Cells

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For all time-lapse studies, the cells were plated in a polymer-coverslip bottom µ-Dish 35 mm (ibidi #81156). THP-1 cells (2×10⁶) were differentiated for 48 h and then transduced with CPSF6 mNeonGreen LV (MOI = 0.01) for 3 days. Next, the cells were infected with HIV-1 and the live imaging was performed during 24–72 h p.i. For CPSF6 cluster fusion studies, CPSF6 mNeonGreen-positive cells were imaged every 5 min in 2D with a Biostation IM-Q (Nikon).
For FRAP experiments, the selected ROI was irradiated for 200 µs/pixel with 488 nm/561 nm laser and, after bleaching, the frames were acquired every 5–10 sec for 5 min through a Ti2E inverted microscope (Nikon), based on a CSU-W1 spinning-disk (Yokogawa), using a 60× objective (Plan Apochromat, oil immersion, NA = 1.4).
Experiments of live tracking of GIR and vDNA (OR-GFP) were performed in differentiated THP-1 cells transduced with OR-GFP LV (MOI = 5). Two days post-transduction, the cells were infected with HIV-1 ANCH3 GIR virus (MOI = 30). Different cells were imaged during 45–96 h p.i., every 5 min in 3D (stacks spacing 0.3 µm) with a Ti2E inverted microscope (Nikon), based on a CSU-W1 spinning-disk (Yokogawa), using a 60× objective (Plan Apochromat, oil immersion, NA = 1.4).

Scoca V., Morin R., Collard M., Tinevez J.Y, & Di Nunzio F. (2022). HIV-induced membraneless organelles orchestrate post-nuclear entry steps. Journal of Molecular Cell Biology, 14(11), mjac060.

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Live/Dead Cell Viability Assay

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Cell viability was assessed using a live/dead viability/cytotoxicity assay (L3224, Invitrogen, Carlsbad, CA, USA). After aspiration of all four media reservoirs of each device, 150 µl of staining solution prepared from 5 ml of EGM-2, 10 µl of ethidium homodimer-1 (Etdh-1, detecting dead cells; Invitrogen), 2.5 µl of Calcein-AM (detecting live cells; Invitrogen), and 5 µl of Hoechst (detecting nuclei; Molecular Probes, Eugene, OR, USA) was pipetted into the media reservoirs. The devices were then incubated at 37°C in 5% CO₂ for 45 min. Fluorescence images were obtained using a confocal microscope (Nikon Ti2-E inverted microscope).

Ahn J., Yoon M.J., Hong S.H., Cha H., Lee D., Koo H.S., Ko J.E., Lee J., Oh S., Jeon N.L, & Kang Y.J. (2021). Three-dimensional microengineered vascularised endometrium-on-a-chip. Human Reproduction (Oxford, England), 36(10), 2720-2731.

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Super-resolution Imaging of Biopsy and Cell Samples

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For patient biopsy samples, imaging was performed on a 3D N-STORM super-resolution microscope (Nikon, Minato City, Tokyo, Japan), consisting of a Ti-E inverted microscope with an E-TIRF illuminator and a Piezo Z stage on a vibration isolation table. This N-STORM system was equipped with a CFI 100XH oil objective Apo, TIRF, NA 1.49, N-STORM and λ/4 lenses, quad cube C-NSTORM (Chroma, Cat# 97355, Bellows Falls, VT, USA), a Perfect Focus System, an MLC-MBP-ND laser launch (Agilent, Santa Clara, CA, USA), and an iXON Ultra DU-897U EMCCD camera (Andor Technology, Belfast, UK). For cultured cell lines, we used an upgraded N-STORM system with a Nikon Ti2-E inverted microscope, Nikon N-STORM TIRF illuminator with a 2× magnification lens, and Nikon LU-NF laser launch. The laser powers used from the Ti-E and Ti2-E systems to excite the AF647 dyes were 140 mW and 89 mW, respectively, as measured out of the optical fiber. The laser power for the activation of AF405 dye was 5–10 mW, as measured out of the optical fiber. We acquired 60,000 frames for the cultured cell lines and 20,000 frames for the patient biopsy samples using an exposure of 10 ms. The data were acquired using NIS Elements software (Nikon, Version 4.3 for Ti-E and 5.21.01 for Ti2-E).

Maddox A.L., Brehove M.S., Eliato K.R., Saftics A., Romano E., Press M.F., Mortimer J., Jones V., Schmolze D., Seewaldt V.L, & Jovanovic-Talisman T. (2022). Molecular Assessment of HER2 to Identify Signatures Associated with Therapy Response in HER2-Positive Breast Cancer. Cancers, 14(11), 2795.

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Fixed and Live-cell Imaging Setup

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Fixed and live-cell images were acquired using a Ti-E or Ti-2E inverted microscope (Nikon Instruments) driven by NIS Elements software (Nikon Instruments). Images were captured using a Clara cooled charge-coupled device (CCD) camera (Andor) or Prime Bsi sCMOS camera (Teledyne Photometrics) with a Spectra-X light engine (Lumencore). For live-cell imaging, cells in CO₂-independent media (Gibco) were imaged using Nikon objectives Plan Apo 20 ×0.75 NA or 40 ×0.95 NA and an environmental chamber at 37 °C. Fixed-cell images were taken using Plan Apo 40 ×0.95 NA, Plan Apo λ 60 ×1.42 NA, and APO 100 ×1.49 NA (Nikon).

Marquis C., Fonseca C.L., Queen K.A., Wood L., Vandal S.E., Malaby H.L., Clayton J.E, & Stumpff J. (2021). Chromosomally unstable tumor cells specifically require KIF18A for proliferation. Nature Communications, 12, 1213.

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Rat Hippocampal Neurons Imaged over Time

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Primary rat hippocampal E18 neurons were plated on 25mm round glass coverslips (Warner Instruments) at a density of 4x10⁵ cells per coverslip. Cells were maintained as described above. Neurons were transfected at DIV14 with Lifeact-GFP using Lipofectamine 2000 (Invitrogen) according to manufacturer instructions. Neurons were imaged with a 60X oil immersion objective (Nikon Plan Apo, N.A. 1.40) on a Nikon (Tokyo, Japan) Ti2-E inverted microscope with a SOLA light source. Their environment was maintained with a Tokai Hit stage top incubation system, with settings as follows: Top heater 42.3°C; Stage Heater 38.3°C; Bath Heater 41°C; Lens Heater 41°C; CO₂ concentration 5%. Neurons were imaged with the following parameters: SOLA light source, 10%; exposure, 200 ms; image size, 1028 × 1028 pixels. Images were captured with an ORCA-Flash 4.0 V3 CMOS camera (Hamamatsu, Hamamatsu City, Japan). An image was captured every 15 minutes for a total of 6 hours. DMSO, 500 nM Aβ42 and/or 10 μm SR7826 were added after the first two images were acquired. For spine density analysis, spines from a representative secondary dendrite at least 25 μm from the soma were counted at each time point and plotted over time.

Henderson B.W., Greathouse K.M., Ramdas R., Walker C.K., Rao T.C., Bach S.V., Curtis K.A., Day J.J., Mattheyses A.L, & Herskowitz J.H. (2019). Pharmacologic inhibition of LIMK1 provides dendritic spine resilience against amyloid-β. Science signaling, 12(587), eaaw9318.

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Fluorescent Microgel Size Characterization

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For size characterization of individual populations of microgels with fluorescence microscopy, PEG-thioester-norbornene, PEG-ester-norbornene, and PEG-amide-norbornene microgels were labeled with 20 μM Alexa Fluor 546 C (Invitrogen), 20 μM Dylight 405 maleimide (Invitrogen), 20 μM and fluorescein-maleimide (Sigma), respectively. PEG-MMP microgels were labeled with 20 μM fluorescein-maleimide for size characterization. Maleimide moieties on each fluorophore react with PEG thiol via thiol-Michael addition. These fluorescent labels were mixed with the respective polymer precursor solution before electrospraying. Fluorescent PEG microgels were suspended in PBS in a multiwell glass-bottom plate and imaged on a Nikon Ti2-E inverted microscope. Images were corrected for bleaching using histogram matching and converted to binary using a local thresholding algorithm. Microgels were selected, segmented, and diameters and roundness were measured in FIJI. PDI is calculated as (standard deviation / mean)².

Miksch C.E., Skillin N.P., Kirkpatrick B.E., Hach G.K., Rao V.V., White T.J, & Anseth K.S. (2022). 4D Printing of Extrudable and Degradable Poly(Ethylene Glycol) Microgel Scaffolds for Multidimensional Cell Culture. Small (Weinheim an der Bergstrasse, Germany), 18(36), e2200951.

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Assessing Apoptosis in Transfected Cells

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Cells were seeded at 15,000 cell/well onto IbiTreat 8-well chambers and allowed to adhere for 24 h. They were subsequently transfected with sfGFP, TCP-VASH1, or TCP-VASH2 plasmids for 24 h. The cells were then fixed, permeabilized and stained as described above with a Cleaved Caspase-3 antibody and an Alexa Fluor 594 secondary antibody. Images were acquired on a Nikon Ti2-E inverted microscope at 10× magnification. To analyze these images, first, the boundary of a successfully transfected cell was identified using the GFP channel in Nikon HCA to create an object for each cell. The same metrics to identify GFP+ objects were utilized as from the imaging of transfected live cells. With each object identified, the boundary of the object was slightly eroded to ensure that the AF549 intensity associated with Caspase-3 is only a result of that cell, and not any adjacent cells or debris. The average AF594 intensity of the cell is then calculated for each eroded object. Cells were determined to be positive for active Caspase-3 when the average 594 intensity of the GFP+ object was calculated to be >10,000 RFUs.

Mathias T.J., Ju J.A., Lee R.M., Thompson K.N., Mull M.L., Annis D.A., Chang K.T., Ory E.C., Stemberger M.B., Hotta T., Ohi R., Vitolo M.I., Moutin M.J, & Martin S.S. (2022). Tubulin Carboxypeptidase Activity Promotes Focal Gelatin Degradation in Breast Tumor Cells and Induces Apoptosis in Breast Epithelial Cells That Is Overcome by Oncogenic Signaling. Cancers, 14(7), 1707.

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Transwell Migration Assay for BMMSCs

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The experimentally-treated Transwell chambers were fixed (anhydrous methanol: Glacial acetic acid 3:1) for 30 min, stained with a 2% crystal violet dye solution for 30 min and washed with phosphate-buffered saline (PBS). The upper layer of cells was removed with a cotton swab, peeled off and placed on a slide, fixed with neutral gum, and then observed under a Ti2-E inverted microscope, (Nikon Corporation). The TNF-α/IEC-6 cells were prepared in a 35-mm diameter well and added to a Transwell chamber containing Ad/MSCs, Ad-CXCR3,/MSCs or Ad-(CXCR3 + HO)/MSCs. The green fluorescent protein (GFP) signal was locked with a living cell workstation microscope, and GFP-expressing BMMSCs located 5 µm below the Transwell membrane were observed to record the time at which when BMMSCs began to appear.

Yin M., Shen Z., Yang L., Zheng W, & Song H. (2019). Protective effects of CXCR3/HO-1 gene-modified BMMSCs on damaged intestinal epithelial cells: Role of the p38-MAPK signaling pathway. International Journal of Molecular Medicine, 43(5), 2086-2102.

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Super-Resolution Imaging of Cellular Structures

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SIM images were taken with an N-SIM system (Nikon) attached to a Ti2-E inverted microscope (Nikon) with a CMOS camera (ORCA-Flash 4.0 V3; Hamamatsu Photonics) using a Plan Apochromat 100×/1.35 NA silicon-immersion objective lens at a step size of 0.12 μm. The chromatic aberration of the system was calibrated and corrected by 0.1 μm TetraSpeck beads (T7279; Thermo Fisher Scientific) in the mounting medium. Images were reconstructed and analyzed using NIS elements AR (Nikon) according to the manufacturer's protocol.

Tocan V., Hayase J., Kamakura S., Kohda A., Ohga S., Kohjima M, & Sumimoto H. (2021). Hepatocyte polarity establishment and apical lumen formation are organized by Par3, Cdc42, and aPKC in conjunction with Lgl. The Journal of Biological Chemistry, 297(6), 101354.

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Quantitative Immunofluorescence Microscopy Analysis

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Immunofluorescence microscopy was performed with a Nikon Ti2-E inverted microscope equipped with Spinning Disk Super Resolution by Optical Pixel Reassignment Microscope (Yokogawa CSU-W1 SoRa, Nikon) and Microlens-enhanced Nipkow Disk with pinholes and 60x SR Plan Apo IR oil immersion objective. Images were acquired at room temperature. Cells were grown on 12 mm coverslips (GmbH & Co KG), washed in PBS and fixed by adding 4 % PFA in 0.1 M sodium phosphate. Cells were fixed at room temperature for 25 min and then washed 3X with PBS. Samples were permeabilized by immersing coverslips in ice-cold methanol for 2–3 s, followed by PBS rinses. Samples were then blocked in PBS-1X with 5% normal donkey serum (Jackson) for 1 h at RT. Subsequent antibody incubations were performed in this buffer. Antibodies used in this study are listed in Table 5.
Images were processed with ImageJ/FIJI (Schindelin et al., 2012 (link)). For analysis of protein colocalization, Manders’ coefficient was determined with ImageJ/Fiji through JACoP plugin (Bolte and Cordelieres, 2006 (link)). For lysosome size measurements, lysosome perimeter was determined with ImageJ/Fiji through StarDist plugin (Schmidt et al., 2018 ). Thresholds were the same for all images analyzed in a given experiment.

Talaia G., Bentley-DeSousa A, & Ferguson S.M. (2023). Lysosomal TBK1 Responds to Amino Acid Availability to Relieve Rab7-Dependent mTORC1 Inhibition. bioRxiv.

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