Elipse ti

Manufactured by Nikon

The Elipse Ti is a high-performance laboratory microscope designed by Nikon. It features a sturdy, ergonomic design and advanced optics to provide clear, detailed images for a variety of scientific applications.

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Lab products found in correlation

Vybrant cell adhesion assay kit, by Thermo Fisher Scientific (4 mentions) Glomax discover system, by Promega (4 mentions) Scmos orca flash 4, by Hamamatsu Photonics (2 mentions) X60 1.42 na apo tirf oil immersion objective, by Nikon (2 mentions) Csu x1 spinning disk unit, by Yokogawa (1 mentions) Csu x1 spinning disk unit, by Nikon (1 mentions) Dmi8 microscope, by Leica camera (1 mentions) Edu assay kit, by Thermo Fisher Scientific (1 mentions) Dynabead, by Thermo Fisher Scientific (1 mentions) Emccd sensor, by Nikon (1 mentions) Nis element, by Nikon (1 mentions) D eclipse c1, by Nikon (1 mentions) Ixon ultra camera, by Nikon (1 mentions) Prism 9, by GraphPad (1 mentions) Oil objective, by Zeiss (1 mentions) Axio imager z2 upright microscope, by Zeiss (1 mentions) Clara interline ccd camera, by Oxford Instruments (1 mentions) Plan apo 60 oil objective, by Nikon (1 mentions) Mineral oil, by Merck Group (1 mentions) 96 well flat bottomed plate, by Corning (1 mentions)

12 protocols using elipse ti

Imaging Third Instar Larval Brains

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Third instar larval brains (120 to 144h after egg lying depending on overall growth delay) were dissected in Schneider medium containing 10% FCS and transferred to 50 μL wells (Ibidi, μ-Slide Angiogenesis) for live imaging. Mutant and control brains were imaged in parallel at 25°C. Z-series (thickness of 20 μm with 1 μm spacing) were acquired with a temporal resolution of 30 to 90 s for 1 to 2.5 hours. Alternatively, samples were mounted on a stainless-steel slide, between coverslip and mineral oil as described in a previous study [59 (link)].
Images were acquired with a spinning disk system consisting of a DMi8 microscope (Leica) equipped with a 63X (1.4 N.A.) oil objective, a CSU-X1 spinning disk unit (Yokogawa) and an Evolve EMCCD camera (Photometrics). The microscope was controlled by the Inscoper Imaging Suite and the dedicated software (Inscoper). Alternatively, a CSU-X1 spinning-disk unit mounted on an inverted microscope (Elipse Ti; Nikon) equipped with a 60X (1.4 N.A.) oil objective, a sCMOS ORCA Flash 4.0 (Hamamatsu) and controlled by MetaMorph; was also used for some experiments. Images were processed with Fiji or Imaris softwares.

Gallaud E., Richard-Parpaillon L., Bataillé L., Pascal A., Métivier M., Archambault V, & Giet R. (2022). The spindle assembly checkpoint and the spatial activation of Polo kinase determine the duration of cell division and prevent tumor formation. PLoS Genetics, 18(4), e1010145.

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Tracking Exocytosis in COS-1 Cells

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COS-1 cells were plated at low density on glass coverslips and transfected with APP-SEP (super ecliptic pHluorin) and with pARIS HTT or pARIS HTT_SA (Pardo et al., 2010 (link)) using calcium phosphate. Acquisitions were made the day after transfection at 5 Hz during 1 min using an inverted microscope (Elipse Ti, Nikon) with a X60 1.42 NA APO TIRF oil-immersion objective (Nikon) coupled to a CCD camera (CoolSnap, Photometrics) and maintained at 37°C and 5% CO2. Analysis was done on area delimited by cell edges and exocytosis rate was quantified using ExocytosisAnalyser macro on ImageJ developed by Marine Scoazec.

Bruyère J., Abada Y.S., Vitet H., Fontaine G., Deloulme J.C., Cès A., Denarier E., Pernet-Gallay K., Andrieux A., Humbert S., Potier M.C., Delatour B, & Saudou F. (2020). Presynaptic APP levels and synaptic homeostasis are regulated by Akt phosphorylation of huntingtin. eLife, 9, e56371.

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Assessing Prostate Cancer Cell Proliferation

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5,000 to 7,500 cells were plated in microculture devices as previously described (32 (link)). LNCaP or LNCaP-C4 cells were cultured with either UGSM-2, UGSM-2 + exogenous sonic hedgehog (Shh, Curis), or Gli3-/- UGSM cells (UGli3^-./-)(33 (link)). Drugs were added on day 1 and replenished until day 5. Following the growth period, cell proliferation was quantified using an EdU assay kit (Invitrogen) or RNA was extracted using DynaBeads (Invitrogen) (33 (link)). EdU assay images were obtained on an inverted Nikon Elipse Ti using a 4x objective. Fluorescent nuclear counts and GPF intensities were determined using ImageJ v1.38 (NIH). % EdU (+) cells were obtained by dividing total EdU (+) cells to total cell number (Hoescht-nuclear stain) X 100. Cell proliferation was assessed for significant differences by Wilcoxon Mann-Whitney test. Significant differences have a p-value <0.05.

Powers G.L., Hammer K.D., Domenech M., Frantskevich K., Malinowski R.L., Bushman W., Beebe D.J, & Marker P.C. (2014). Phosphodiesterase 4D Inhibitors Limit Prostate Cancer Growth Potential. Molecular cancer research : MCR, 13(1), 149-160.

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Monocyte Adhesion to Endothelial Cells

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The adhesion of monocytes (THP-1 cells) to EA.hy926 cells was determined using the Vybrant Cell Adhesion Assay Kit (ThermoFisher Scientific). EA.hy926 cells were seeded in 96-well flat bottom plates (density of 2 × 10⁵ cells/mL, 1 × 10⁵ cell per well). After incubation with FAs for 48 h and then with TNF-α for 24 h, calcein-labelled THP-1 cells (5 × 10⁴ cells in 100 µL) were incubated with EA.hy926 cells for 1 h at 37 °C. Non-adherent THP-1 cells were removed by gentle washing, 100 µL PBS added to each well and co-cultures read on the Glomax Discover System (Promega). THP-1 monocyte adhesion was measured as a percentage of control (non-stimulated DMEM treated cells). Images of fluorescence-labelled THP-1 monocytes bound to EA.hy926 cells were taken with a Nikon Elipse Ti using NIS elements software (version 4.30).

Valenzuela C.A., Baker E.J., De Souza C.O., Miles E.A, & Calder P.C. (2021). Differential Effects of Ruminant and Industrial 18-Carbon trans-Monounsaturated Fatty Acids (trans Vaccenic and Elaidic) on the Inflammatory Responses of an Endothelial Cell Line. Molecules, 26(19), 5834.

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Live-cell TIRF Microscopy for Puncta Analysis

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TIRF imaging was as we described before (Jha et al., 2019 (link)). Images were recorded at 37°C with Nikon NIS-Elements paired with the Nikon Elipse Ti with PFS (Perfect Focus System) autofocus capabilities, Nikon N-Storm, Andor iXon Ultra Camera with EMCCD Sensor, D-Eclipse C1, and 60× TIRF objective lens (Nikon), 1.45 Na⁺ Oil immersion, infinity/0.10–0.22 DIC H. The size and intensity of the puncta were analyzed by ImageJ in imported images recorded by the Nikon NIS software. Background was subtracted from the first image in a sequence and maintained through the time course. The area of the cell from which the puncta were analyzed was determined by the NIS-elements software, and number and intensity were normalized using this area. The results are shown as mean ± SEM, and statistical analysis and figures were made with GraphPad Prism 9.

Liu H., Lin W.Y., Leibow S.R., Morateck A.J., Ahuja M, & Muallem S. (2022). TRPC3 channel gating by lipids requires localization at the ER/PM junctions defined by STIM1. The Journal of Cell Biology, 221(5), e202107120.

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Fluorescence Imaging of Cell Structures

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Bright-field images were acquired on an Axio Imager.Z2 upright microscope (Zeiss) running AxioVision LE64, release 4.9.1 SP1 (Zeiss) for whole mount, in situ hybridization and lacZ staining. Immunofluorescence and PFO* images were acquired on either an Elipse Ti with a CSU-X1 spinning disc confocal (Nikon) and Clara interline CCD camera (Andor) with a Plan Apo ×60 oil objective (Nikon) running Nikon Elements 5.02 build 1266 (Nikon) or an LSM 800 confocal laser scanning microscope with a ×63 oil objective (Zeiss) running Zen 2 blue edition v1.0 (Zeiss). Cell length calculations, SMO intensity analysis, and PFO* intensity at the primary cilium analysis were done in FIJI.

Lo M., Sharir A., Paul M.D., Torosyan H., Agnew C., Li A., Neben C., Marangoni P., Xu L., Raleigh D.R., Jura N, & Klein O.D. (2022). CNPY4 inhibits the Hedgehog pathway by modulating membrane sterol lipids. Nature Communications, 13, 2407.

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Immunofluorescence Staining of Tubulin in Monolayer Cells

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Monolayer culture was performed by seeding cell on a glass bottom culture dish with 60% confluence. After that, cells were prepared for immunofluorescence staining according to standard protocols. Cells were fixed in cold 4% paraformaldehyde for 20 min, washed 3 times in 0.5% Triton X-100 in PBS (PBST) for 5 min, and incubated in blocking solution consisting of 1% bovine serum albumin (BSA) in PBST for 2 h. Anti-alpha tubulin (Abbkine, diluted 1:200) and DyLight549 goat anti-mouse (Abbkine, diluted 1:200) antibodies were used as the primary antibody and secondary antibody, respectively. 4′,6-diamidino-2-phenylindole (DAPI) was used for nuclear staining. A laser confocal microscope (NIKON Elipse Ti) was used for observation [16 (link), 17 (link)].

Gao E.M., Turathum B., Wang L., Zhang D., Liu Y.B., Tang R.X, & Chian R.C. (2021). The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles. Reproductive Sciences, 29(4), 1343-1356.

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Monocyte Adhesion to Endothelial Cells

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Adhesion of monocytic THP-1 cells to EA.hy926 cells was determined using the Vybrant Cell Adhesion Assay Kit (ThermoFisher Scientific). EA.hy926 cells were exposed to FAs at 25 and 50 µM for 48 hr followed by stimulation with TNF-α (1 ng/mL) for 6 hr. After stimulation, calcein labelled THP-1 cells were incubated with EA.hy926 cells for 1 hr at 37 o C. Nonadherent THP-1 cells were removed by gentle washing, 100 µL PBS added to each well and co-cultures read on the Glomax Discover System (Promega). THP-1 monocyte adhesion was measured as a percentage of control (TNFα stimulated cells). Images of fluorescencelabelled THP-1 monocytes bound to EA.hy926 cells were taken with a Nikon Elipse Ti using NIS elements software (version 4.30).

Baker E.J., Valenzuela C.A., De Souza C.O., Yaqoob P., Miles E.A, & Calder P.C. (2020). Comparative anti-inflammatory effects of plant- and marine-derived omega-3 fatty acids explored in an endothelial cell line. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1865(6).

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Visualizing APP Exocytosis in COS-1 Cells

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COS-1 cells were plated at low density on glass coverslips and transfected with APP-SEP (super ecliptic pHluorin) and with pARIS HTT or pARIS HTT SA (Pardo et al., 2010) using calcium phosphate.
Acquisitions were made the day after transfection at 5Hz during 1 min using an inverted microscope (Elipse Ti, Nikon) with a X60 1.42 NA APO TIRF oil-immersion objective (Nikon) coupled to a CCD camera (CoolSnap, Photometrics) and maintained at 37°C and 5% CO2. Analysis was done on area delimited by cell edges and exocytosis rate was quantified using ExocytosisAnalyser macro on ImageJ developed by Marine Scoazec.

Bruyère J., Abada Y., Vitet H., Fontaine G., Deloulme J., Cès A., Denarier É., Pernet‐Gallay K., Andrieux A., Humbert S., Potier M., Delzescaux T., & Saudou F. (2020). Presynaptic APP levels and synaptic homeostasis are regulated by Akt phosphorylation of Huntingtin.

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Live Imaging of Microtubule Dynamics

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Third-instar larval brains were dissected in Schneider's Drosophila medium supplemented with 10% FCS. Isolated brains were loaded and mounted on stainless steel slides, and the preparations were sealed with mineral oil (Sigma-Aldrich) as previously described 14 . For MT depolymerization experiments, larval brains were incubated during 30 min in the above medium supplemented with colchicine at a final concentration of 15 μM. After incubation, brains were mounted and processed for live cell imaging.
Images were acquired at 25°C using a CSU-X1 spinning-disk system mounted on an inverted microscope (Elipse Ti; Nikon) equipped with a 60X 1.4 NA objective. At 20, 30 or 60 sec intervals 10 z-steps were acquired with 1µm intervals. Fluorescent protein probes were excited with 488nm or 561nm laser light and the images were captured using a sCMOS ORCA-Flash4.0 (Hamamatsu) camera. Recordings were controlled using MetaMorph acquisition software. Data were processed in ImageJ and viewed as maximum-intensity projections prior to analysis or figure preparation.

Thomas A., Gallaud E., Richard‐Parpaillon L., Serre L., Arnal I., Richard‐Parpaillon L., Savoian M.S., & Giet R. (2020). Peripheral microtubules ensure asymmetric furrow positioning in neural stem cells.

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