Ti e microscope

Manufactured by Yokogawa

Sourced in Japan

The Ti-E microscope is a high-performance inverted research microscope designed for a wide range of applications. It features a motorized nosepiece, condenser, and focus drive for precise and efficient sample observation. The Ti-E provides excellent optical performance and flexibility to accommodate various accessories and imaging techniques.

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Lab products found in correlation

Csu21, by Yokogawa (3 mentions) Csu w1 spinning disk confocal unit, by Oxford Instruments (3 mentions) Alc600 laser beam combiner, by Oxford Instruments (3 mentions) C9100 50 emccd camera, by Hamamatsu Photonics (2 mentions) Csu x1 scan head, by Hamamatsu Photonics (2 mentions) Ixon ultra 888 emccd camera, by Oxford Instruments (2 mentions) Perfect focus system, by Nikon (2 mentions) Csu x1 spinning disk unit, by Yokogawa (2 mentions) Nis elements software, by Nikon (2 mentions) Csu w1 spinning disk head, by Yokogawa (1 mentions) Flash 4 scmos camera, by Hamamatsu Photonics (1 mentions) Csu w1, by Hamamatsu Photonics (1 mentions) Orca flash4.0 v2 scmos camera, by Hamamatsu Photonics (1 mentions) Csu 10 spinning disk head, by Hamamatsu Photonics (1 mentions) Imageem emccd camera, by Hamamatsu Photonics (1 mentions) Du897 ultra emccd camera, by Oxford Instruments (1 mentions) Halocarbon oil 27, by Merck Group (1 mentions) Sp5 confocal system, by Leica (1 mentions) Sir dna, by Spirochrome (1 mentions) Perfusion chamber, by Warner Instruments (1 mentions)

Close spelling variants of this product

Ti-E inverted microscope (19 citations) Ti microscope (13 citations) Eclipse Ti-E inverted microscope (6 citations) Eclipse Ti-E microscope (5 citations) Eclipse Ti-E base microscope (3 citations) Ti-E-PFS inverted microscope (2 citations)

24 protocols using ti e microscope

Observing Root Epidermal Cell Expansion

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Timing: 5–6 h

These steps aim to observe root epidermal cell expansion during root bending.

45.

Pour 1/2 MS medium into a chambered cover slide (ibidi, 80287), and make sure to fill the entire chamber by covering it with a sterile glass plate.

Note: For halo-stimulation, replace the bottom left part with 1/2 MS medium containing 200 mM NaCl. For the control treatment, replace the bottom left part with 1/2 MS medium.

46.

Install a 20× objective onto the objective inverter (LSM TECH), and turn on the spinning-disk confocal microscope.

Note: The spinning-disk confocal microscopy is an inverted Nikon Ti-E microscope with a CSU-W1 spinning disk head (Yokogawa, Japan), with a deep-cooled evolve charge-coupled iXon Ultra 888 EM-CCD camera (Photometrics Technology, USA).

47.

Transfer a 4-day-old seedling with straight root onto the split-agar medium in the chambered cover slide, cover the seedling with a cover slide, and seal it with micropore tape.

48.

Place the chambered cover slide vertically on the vertical stage.

49.

Adjust to get a clear view of the epidermal cells in the root tip.

50.

Start imaging with Z-steps of 0.5 μm intervals covering a depth of more than 10 μm from the root surface to the interior and 10 min time intervals for 3 h.

Note: A 514-nm laser and a 540/50-nm emission filter are used to obtain the YFP signal.

Yu B., Zheng W., Persson S, & Zhao Y. (2023). Protocol for analyzing root halotropism using split-agar system in Arabidopsis thaliana. STAR Protocols, 4(2), 102157.

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Imaging Tissue Sections with Spinning Disk Confocal

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Tissue sections on the slides were imaged using the Nikon Ti-E microscope coupled to a Yokogawa CSU-W1 spinning disk using the Hamamatsu Flash 4 sCMOS camera. The images were captured using 60× Plan Apochromat (NA 1.4) oil objective. The sections to image were manually identified using a 10× objective (NA 1.45). The images were obtained at 100% laser power for far red 633 nm, red 561 nm, green 488 nm and DAPI 405 nm lasers. For each channel the following filters were used: DAPI, ET455/50m; green, ET525/36m; red, ET605/70m; far red, ET700/75m. A Nikon Elements Job ‘Tiler’ was used to capture the images if tiles were taken, and the image was stitched later in Fiji (https://imagej.net/software/fiji) using the grid/collection stitching plugin. The order of experiments was Lambda (z-series), so each color was imaged in z before moving to the next color. For obtaining a z-stack through the tissue section, each slice imaged was 1 µm apart and a range of 20 steps was taken. Images were obtained in the order of 633 (for the 647 probe), 561 (for the 555 probe), 488 (for the atto 488 probe) and 405 (for DAPI). On a slide, the whole tissue section was imaged, and for each slide a single row was imaged all the way across for each genotype of embryo. All images were stitched using Fiji before further processing.

Afzal Z., Lange J.J., Nolte C., McKinney S., Wood C., Paulson A., De Kumar B., Unruh J., Slaughter B.D, & Krumlauf R. (2023). Shared retinoic acid responsive enhancers coordinately regulate nascent transcription of Hoxb coding and non-coding RNAs in the developing mouse neural tube. Development (Cambridge, England), 150(10), dev201259.

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Visualizing cortical and embryonic microtubule networks

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Gravid adults were dissected directly on polylysine-coated
coverslips in a drop of egg buffer and were gently flattened by mounting
with 22.8 μm beads (Whitehouse scientific, Chester, UK) as spacers.
Cortical images were acquired using a Nikon Ti-E microscope equipped with a
100X, 1.49 NA objective; a Yokogawa CSU-X1 spinning disk head; and a
Hamamatsu ImageEM EM-CCD camera. Embryo cross-section images were acquired
using a Nikon TE-2000 microscope equipped with a 60X, 1.4 NA objective; a
Yokogawa CSU-10 spinning disk head; and a Hamamatsu Orca-Flash4.0
V2+ scMOS camera. For embryo imaging, mNG was excited using a 40 mW,
514 nm laser, which produced less autofluorescent background and
phototoxicity compared to 488 nm. GFP was excited using a 50 mW, 488 nm
laser and mKate2 was excited using a 50 mW, 561 nm laser.

Dickinson D.J., Schwager F., Pintard L., Gotta M, & Goldstein B. (2017). A single-cell biochemistry approach reveals PAR complex dynamics during cell polarization. Developmental cell, 42(4), 416-434.e11.

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Live Cell Fluorescence Microscopy of S. pombe

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All S. pombe fluorescence microscopy was performed on a Nikon Ti-E microscope with a CSI W1 spinning disk (Yokogawa) using a 100 ×/ 1.4 NA Olympus Plan Apo oil objective and an iXon DU897 Ultra EMCCD camera (Andor). Fluorophores were excited at 488 nm (GFP) and 561 nm (mCherry) and collected through an ET525/36 m (GFP) or ET605/70 m (mCherry) bandpass filter. Images were collected over a z-volume of 8 µm with 0.5 µm spacing for 4–5 h at 5-min intervals. Image acquisition conditions for all experiments are listed in Table S4. Live imaging was performed in 35-mm glass bottom dishes (MaTek, no. 1.5 coverslip) maintained at 25°C using an Oko Lab stage top incubator.
For imaging meiotic divisions, strains were prepared as specified above. For imaging mitotic divisions, strains were grown in yeast extract with supplements (YES; 5 g yeast extract, 30 g dextrose, 0.2 g each adenine, uracil, histidine, leucine, and lysine, in 1 L of water) and maintained in exponential growth for at least 2 d prior to imaging. The 35-mm glass bottom dishes were coated with 1 mg/ml soybean lectin (in water) for 15 min, rinsed with 1 ml YES media, and 200 µl of log phase cell culture was added and allowed to settle for 30 min at 30°C. Cells were rinsed twice with prewarmed YES media and then imaged.

King G.A., Wettstein R., Varberg J.M., Chetlapalli K., Walsh M.E., Gillet L.C., Hernández-Armenta C., Beltrao P., Aebersold R., Jaspersen S.L., Matos J, & Ünal E. (2022). Meiotic nuclear pore complex remodeling provides key insights into nuclear basket organization. The Journal of Cell Biology, 222(2), e202204039.

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Visualizing Drosophila Embryo Nuclei Dynamics

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Embryos were dechorionated by gently rolling them on a double-sided adhesive tape until the chorion layer was completely peeled. Then, embryos were mounted in a halocarbon oil 27 (Sigma) on a gas-permeable membrane (aka bio-foil; Kenneth Technologies), and covered with a high-definition 1.5H coverslip (Paul Marienfeld). Images were acquired 5–10 μm below the visible apical end of the cells on a Leica SP5 confocal system with an HCX PL APO CS 63×/1.4-NA oil-immersion objective. Time-lapses were acquired at 561 nm excitation wavelength, line averaging with 1–3 repeats, frame taken every 29–32 seconds, and pixel size 0.2405 μm². Embryo for live image of nuclear cycles (S3 File) was dechorionated with 50% bleach, washed and mounted on Biofoil membrane in halocarbon 27 oil. The embryo was then imaged on the Nikon Ti-E microscope with the Yokogawa spinning disc (CSU-21) module using the 561 laser to visualize nuclei at 10 s intervals.

Stern T., Shvartsman S.Y, & Wieschaus E.F. (2020). Template-based mapping of dynamic motifs in tissue morphogenesis. PLoS Computational Biology, 16(8), e1008049.

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Visualizing Nuclei Dynamics in Fly Embryos

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Embryos were collected from either wild type or flies maternally expressing RNRL^D68N that also expressed H2AV::mRFP in order to visualize nuclei. Embryos were dechorionated with 50% bleach, washed and mounted on Biofoil membrane in halocarbon 27 oil. Embryos were then imaged on the Nikon Ti-E microscope with the Yokogawa spinning disc (CSU-21) module using the 561 laser to visualize nuclei at 10 second intervals.

Djabrayan N.J., Smits C.M., Krajnc M., Stern T., Yamada S., Lemon W.C., Keller P.J., Rushlow C.A, & Shvartsman S.Y. (2019). Metabolic regulation of developmental cell cycles and zygotic transcription. Current biology : CB, 29(7), 1193-1198.e5.

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Fluorescence Imaging of C. elegans Synaptic Vesicles

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Raw data collection was performed as previously described^{7 (link)}. Briefly, young-adult hermaphrodite C.elegans expressing the synaptic vesicle precursor marker RAB-3 in the DA9 motor neuron (wyIs251[Pmig-13::GFP::RAB-3]) were paralyzed in 0.3 mM Levamisole in M9. Once paralyzed, worms were carefully transferred to M9 solution on a 10% agarose pad for imaging. This results in an effective Levamisole concentration which is significantly lower than the concentration that was suggested to affect axonal transport^{42 (link)}. Worms were maintained on the pad for no more than 20 min, although we confirmed that viability was maintained even after 4 h.
Fluorescence imaging was performed using a Nikon 60 × CFI plan Apo VC, NA 1.4 objective on a Nikon Ti-E microscope equipped with Yokogawa CSU-X1 scan-head and a Hamamatsu C9100-50 EM-CCD camera at a frame rate of 110 ms/frame, and 240 nm per pixel.
Post-analysis fluorescence movies were corrected and analyzed in imageJ. Animal movement was corrected using FIJI plugin StackReg. Kymographs were generated with KymoBuider. Intensity was averaged ± 5 pixels transverse to the kymograph line.

Gramlich M.W., Balseiro-Gómez S., Tabei S.M., Parkes M, & Yogev S. (2021). Distinguishing synaptic vesicle precursor navigation of microtubule ends with a single rate constant model. Scientific Reports, 11, 3444.

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Live-cell Imaging of DNA Dynamics

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For live-cell imaging, cells were plated on poly-L-Lysine-coated glass-bottom 2-well imaging slides (ibidi), allowing to image control and auxin-treated conditions in parallel. For DNA staining cells were incubated in media containing 500 nM SiR-DNA (Spirochrome) for 1 h before imaging. Timelapse acquisitions were carried out on a Nikon TiE microscope equipped with a Yokogawa CSU-W1 spinning disk confocal unit (50 µm pinhole size), an Andor Borealis illumination unit, Andor ALC600 laser beam combiner (405 nm/488 nm/561 nm/640 nm), and Andor IXON 888 Ultra EMCCD camera. The microscope was controlled by software from Nikon (NIS Elements, ver. 5.02.00). Cells were imaged in an environmental chamber maintained at 37 °C with 5% CO₂ (Oko Labs), using a Nikon PlanApo 60x/1.49 NA oil immersion objective and a Perfect Focus System (Nikon). Images were recorded every 15 min for 21 h as z-stacks with two planes and a step size of 6 µm, unbinned and with a pixel size of 217 nm. For excitation of mClover and SiR-DNA, the 488 and 640 nm laser lines were used, respectively. Fiji software (ImageJ 1.51j)^{73 (link)} was used to analyze images.

Cremer M., Brandstetter K., Maiser A., Rao S.S., Schmid V.J., Guirao-Ortiz M., Mitra N., Mamberti S., Klein K.N., Gilbert D.M., Leonhardt H., Cardoso M.C., Aiden E.L., Harz H, & Cremer T. (2020). Cohesin depleted cells rebuild functional nuclear compartments after endomitosis. Nature Communications, 11, 6146.

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Live Cell Traction Force Microscopy

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Live cell traction force measurements were performed on an inverted Nikon Ti-E microscope with a CSU-X confocal scanhead (Yokogawa), laser merge module containing 491, 561 and 642 nm laser lines (Spectral Applied Research) and an HQ2 cooled CCD camera (Roper Scientific). All hardware was controlled via Metamorph acquisition software (MDS Analytical Technologies). Traction force data was obtained at 37°C in a perfusion chamber (Warner Instruments) using a 60x 1.2 NA Plan Apo WI objective (Nikon). Cells were maintained in culture media supplemented with 10 mM HEPES and 30 μl/ml Oxyrase (Oxyrase, Inc.).

Soiné J.R., Brand C.A., Stricker J., Oakes P.W., Gardel M.L, & Schwarz U.S. (2015). Model-based Traction Force Microscopy Reveals Differential Tension in Cellular Actin Bundles. PLoS Computational Biology, 11(3), e1004076.

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Imaging HeLa Cells with Spinning Disk Microscopy

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HeLa cells were imaged with a Nikon Ti-E microscope equipped with a Yokogawa CSU X-1 spinning disk unit, a 60× objective (Plan Apo VC, oil, DIC, NA 1.4), Perfect Focus System and the Nikon NIS elements software. Images were acquired with a Andor iXon 897 EMCCD camera. Photo-activation was achieved with a single pulse of the 440 nm laser light, intensity set to 20%, for 1 s. During photo-activation CFPs were imaged using a 440 nm laser line, a triple dichroic mirror (440, 514, 561 nm), and a 460–500 nm emission filter. RFPs were imaged using a 561 nm laser line, a triple dichroic mirror (405, 488, 561 nm), and a 600–660 nm emission filter.

Mahlandt E.K., Palacios Martínez S., Arts J.J., Tol S., van Buul J.D, & Goedhart J. (2023). Opto-RhoGEFs, an optimized optogenetic toolbox to reversibly control Rho GTPase activity on a global to subcellular scale, enabling precise control over vascular endothelial barrier strength. eLife, 12, RP84364.

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