Ti e pfs

Manufactured by Nikon

The Ti-E-PFS is a high-performance microscope stage from Nikon designed for precise focusing and stable imaging. It features a piezo-driven focusing system for rapid and accurate focus control. The Ti-E-PFS is a core component of Nikon's advanced microscope systems, enabling precise and reliable experimental results.

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Lab products found in correlation

Plan apo lambda, by Nikon (2 mentions) Csu x1 spinning disk unit, by Oxford Instruments (2 mentions) 4 line laser module, by Oxford Instruments (2 mentions) 897 emcdd camera, by Oxford Instruments (2 mentions) 897 emccd camera, by Oxford Instruments (1 mentions) Nis elements, by Adobe (1 mentions) Orca flash 4 camera, by Hamamatsu Photonics (1 mentions)

8 protocols using ti e pfs

AI-182 Fluorescent Imaging of Cancer Cells

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Example 10

This example illustrates imaging of cancer cells using AI-182 as a fluorescent probe.

In these experiments, human carcinoma cells were incubated with AI-182

[Figure (not displayed)]

(5 μM) at 37° C. in the presence of 5% CO₂for 30 min, and examined using a Nikon Ti-E PFS inverted high resolution microscope equipped with a Nikon (Magnification: 20×) Plan APO objective, Prior H117 ProScan flat top linear encoded stage, and Prior Lumen 200PRO illumination system with standard DAPI and FITC filter sets. Results are shown in FIG. 10. Top row: Live Cell Imaging of Human Glioblastoma (U87) Cells Using AI-182. Middle row: Live Cell Imaging of Human Pancreatic Cancer Cells (PANC1) Using AI-182. Bottom row: Live Cell Imaging of Human Pancreatic Cancer Cells (Mia PaCa-2) Using AI-182. Note accumulation of the probe within cells.

All references cited herein are incorporated by reference, each in its entirety. Applicant reserves the right to challenge any conclusions presented by any of the authors of any reference.

US10335504B2. Heterocyclic molecules for biomedical imaging and therapeutic applications (2019-07-02). Washington University [US]. Inventors: G. S. M. Sundaram [US], Jothilingam Sivapackiam [US], Vijay Sharma [US].

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Live Sample Imaging with Confocal Microscopy

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Fluorescent images of live or fixed samples were captured using a Nikon Ti-E-PFS inverted spinning-disk confocal microscope equipped with a 60× 1.4 NA Plan Apo Lambda objective. The system is outfitted with a Yokogawa CSU-X1 spinning disk unit, a self-contained 4-line laser module (excitation at 405, 488, 561, and 640 nm), and Andor iXon 897 EMCCD camera. Confocal fluorescent images and DIC images were acquired and processed using the Nikon NIS-Elements and Adobe Photoshop CS5 software.

Boateng R., Nguyen K.C., Hall D.H., Golden A, & Allen A.K. (2017). Novel functions for the RNA-binding protein ETR-1 in Caenorhabditis elegans reproduction and engulfment of germline apoptotic cell corpses. Developmental biology, 429(1), 306-320.

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Imaging Cytotoxic T Cell Dynamics

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Imaging was performed on an inverted microscope (Nikon Ti-E PFS, Melville NY) equipped with a 60x objective for IRM and TFM, and a 100× objective lens TIRF imaging respectively using a Prime BSI camera (Photometrics, Tucson AZ). Imaging protocols were implemented using Nikon Elements and images were cropped in Fiji before further analysis using MATLAB scripts. For live cell imaging, activated CTLs in L-15 medium were seeded on anti-CD3 coated surfaces equilibrated to 37°C in a stage-top Okolab Incubator (Okolab S. R. L., Pozzuoli, NA, Italy). IRM time-lapse images were acquired every 5 s. TIRF images were acquired every 0.5-1 s for imaging of F-Tractin-EGFP transfected cells and every 1 s for imaging of EGFP-EB3 transfected cells.
Fast imaging of lytic granules: Rapid TIRF imaging of lytic granules was performed using a 100x objective and an electron multiplying charge coupled device (emCCD) camera (Andor iXon 897). Activated CTLs expressing Lamp1-RFP were imaged between 1 to 15 minutes after being added to an anti-CD3-coated coverslip. For each cell, timelapse images were acquired every 100 ms for 100 s (1000 frames).

Pathni A., Özçelikkale A., Rey-Suarez I., Li L., Davis S., Rogers N., Xiao Z, & Upadhyaya A. (2022). Cytotoxic T Lymphocyte Activation Signals Modulate Cytoskeletal Dynamics and Mechanical Force Generation. Frontiers in Immunology, 13, 779888.

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Fluorescence Microscopy of Bacterial Cells

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For standard microscopy, M. xanthus or E. coli cells, exponentially growing cells grown in CYE or LB media, respectively, were washed, concentrated in TPM buffer and mounted on microscope slides covered with a 1.5% TPM agarose pad. The cells were imaged on an automated and inverted epifluorescence microscope TiE‐PFS (Nikon), with a 100 × NA = 1.45 Phase Contrast objective and a camera orca flash 4 (Hamamatsu) at room temperature. Mercury fluorescent lamp with green and red optical filters was used when necessary. Images analysis, pictures and movies were prepared for publication using Fiji (https://fiji.sc/) and Adobe Photoshop. All images presented in the Figures and Figures EV are representative of three biological replicates.

Bautista S., Schmidt V., Guiseppi A., Mauriello E.M., Attia B., Elantak L., Mignot T, & Mercier R. (2022). FrzS acts as a polar beacon to recruit SgmX, a central activator of type IV pili during Myxococcus xanthus motility. The EMBO Journal, 42(1), e111661.

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Fluorescent Worm Imaging Procedure

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All fluorescent strains were treated with appropriate RNAi condition at 24°C for 24 h before imaging. 10 µl of anesthetic (0.1% tricane and 0.01% tetramisole in 1X M9 buffer) was added to a 3% agar pad on a slide and 10–15 live worms were transferred to the drop of anesthetic. A glass coverslip was slowly lowered to cover the samples and the coverslip edges were sealed with nail polish and allowed to dry before imaging. Images were obtained on a Nikon Ti-E-PFS inverted spinning-disk confocal microscope using a 60X 1.4NA Plan Apo Lambda objective. The microscope consists of a Yokowaga CSU-X1 spinning disk unit, a self-contained 4-line laser module (excitation at 405, 488, 561, and 640 nm), and an Andor iXon 897 EMCDD camera. Fluorescence intensities were quantified and image editing done using NIS-elements software.

Fernando L.M., Quesada-Candela C., Murray M., Ugoaru C., Yanowitz J.L, & Allen A.K. (2022). Proteasomal subunit depletions differentially affect germline integrity in C. elegans. Frontiers in Cell and Developmental Biology, 10, 901320.

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Anesthetizing and Imaging Live Worms

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10l of anesthetic (0.1% tricane and 0.01% tetramisole in 1X M9 buffer) was added to a 3% agar pad on a slide and 10-15 live worms were transferred to the drop of anesthetic.
A glass coverslip was slowly lowered to cover the samples and the coverslip edges were sealed with nail polish and allowed to dry before imaging. The images were obtained on a Nikon Ti-E-PFS inverted spinning-disk confocal microscope using a 60x 1.4NA Plan Apo Lambda objective. The microscope consists of a Yokowaga CSU-X1 spinning disk unit, a self-contained 4-line laser module (excitation at 405, 488,561, and 640nm), and an Andor iXon 897 EMCDD camera. Fluorescence intensities were quantified and image editing done using NIS-elements software.

Fernando L.M., Elliot J., & Allen A.K. (2020). The C. elegans proteasome subunit RPN-12 is required for hermaphrodite germline sex determination and oocyte quality.

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Time-lapse Microscopy of Drug Response

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Time-lapse microscopy was performed as previously described (Arora et al., 2017) . In brief, cells were plated on a 96-well plate coated with collagen (1:50 dilution in water) in phenol red-free full growth medium. Approximately 24 hr later, cells were transferred to a Ti-E PFS (Nikon) with a humidified, 37°C chamber with 5% CO 2 , and imaged periodically with a 10X 0.45 NA objective. Drug was added to the medium by pausing the movie, exchanging 50% of the medium in the well with medium containing 2X drug concentration, and restarting the movie. Movie-specific parameters are described below.

Tian C., Yang C., & Spencer S.L. (2020). EllipTrack: A Global-Local Cell-Tracking Pipeline for 2D Fluorescence Time-Lapse Microscopy.

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Live Worm Imaging on Spinning-Disk Confocal

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All fluorescent strains were treated with appropriate RNAi condition at 24°C for 24hrs before imaging. 10µl of anesthetic (0.1% tricane and 0.01% tetramisole in 1X M9 buffer) was added to a 3% agar pad on a slide and 10-15 live worms were transferred to the drop of anesthetic. A glass coverslip was slowly lowered to cover the samples and the coverslip edges were sealed with nail polish and allowed to dry before imaging. Images were obtained on a Nikon Ti-E-PFS inverted spinning-disk confocal microscope using a 60x 1.4NA Plan Apo Lambda objective. The microscope consists of a Yokowaga CSU-X1 spinning disk unit, a self-contained 4-line laser module (excitation at 405, 488, 561, and 640nm), and an Andor iXon 897 EMCDD camera. Fluorescence intensities were quantified and image editing done using NIS-elements software.

Fernando L.M., Quesada‐Candela C., Murray M., Ugoaru C., Yanowitz J.L., & Allen A.K. (2022). Proteasomal subunit depletions differentially affect germline integrity in C. elegans.

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