Eclipse e400 microscope

Manufactured by Nikon

Sourced in Japan, United States, United Kingdom, Canada

The Eclipse E400 is a high-performance microscope designed for laboratory use. It features a sturdy construction, advanced optics, and a range of accessories to support various microscopy applications. The core function of the Eclipse E400 is to provide clear, detailed images of specimens for scientific observation and analysis.

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Lab products found in correlation

Lipidure coat plate a u96, by NOF corporation (15 mentions) Ds 5m camera, by Nikon (5 mentions) Vectashield containing dapi, by Vector Laboratories (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Vectashield, by Vector Laboratories (4 mentions) Ds fi1 camera, by Nikon (4 mentions) Fitc and tritc filters, by Nikon (4 mentions) Dxm1200f, by Nikon (3 mentions) Epi fl filter block n uv 2a, by Nikon (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Anti c5b 9, by Abcam (3 mentions) Digital sight ds u2 camera, by Nikon (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Coolpix camera, by Nikon (3 mentions) Alexafluor 488 conjugated goat anti mouse igg1, by Thermo Fisher Scientific (3 mentions) 5804r centrifuge, by Eppendorf (2 mentions) Alexa fluor conjugated goat anti rabbit or goat anti mouse antibody, by Thermo Fisher Scientific (2 mentions) Neutral buffered formalin, by Merck Group (2 mentions) Elements br software, by Nikon (2 mentions) Boyden chamber, by Merck Group (2 mentions)

225 protocols using eclipse e400 microscope

In Vitro Replication of Toxoplasma Tachyzoites

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Purified live and UV-inactivated tachyzoites were examined for in vitro replication competency using a modification of an established immunofluorescence procedure (D’Angelo, et al., 2009 (link)). Tachyzoites were added to HFF cells growing in 8-chamber slides (Millipore). At 2–9 days post-infection (37°C, 5% CO₂), cell monolayers were rinsed with DPBS, fixed, permeabilized and then immunolabeled with rabbit (Rb) anti-SAG-1 (AbD Serotec, UK) followed by Alexa Fluor 594 goat anti-Rb (red, Life Technologies). DAPI (Invitrogen) for visualizing host cell nuclei was added to the secondary antibody. Stained cells were examined by epifluorescence using a Nikon eclipse E400 microscope with the program MetaVue version 6.2r6. Prior to staining, cells were examined by phase contrast using Axiovert 100 (Carl Zeiss) and images taken with Axiovision Rel 4.8.

Kannan G., Prandovszky E., Steinfeldt C.B., Gressitt K.L., Yang C., Yolken R.H., Severance E.G., Jones-Brando L, & Pletnikov M.V. (2014). One Minute Ultraviolet Exposure Inhibits Toxoplasma gondii Tachyzoite Replication and Cyst Conversion without Diminishing Host Humoral-Mediated Immune Response. Experimental parasitology, 145, 110-117.

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Measuring Caspase-3 Activation by Immunofluorescence

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Activation of caspase-3 was determined by indirect immunofluorescence based on a single cell analysis. Briefly, cells were grown on glass coverslips. 24 h after transfection cells were incubated with TNF (30 ng/ml, 4 h) or MPP⁺ (7.5 μM, 4 h) or STS (1 μM, 5 h). The cells were then fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% goat serum in 0.2% Triton X-100 in PBS for 1 h at room temperature. Fixed cells were incubated with an antibody against activated caspase-3 overnight at 4°C, followed by an incubation with ALEXA555-conjugated secondary antibody for 1 h at room temperature. After extensive washing, cells were mounted onto glass slides and analyzed for active caspase-3 by fluorescence microscopy using a Nikon Eclipse E400 microscope.

Meschede J., Šadić M., Furthmann N., Miedema T., Sehr D.A., Müller-Rischart A.K., Bader V., Berlemann L.A., Pilsl A., Schlierf A., Barkovits K., Kachholz B., Rittinger K., Ikeda F., Marcus K., Schaefer L., Tatzelt J, & Winklhofer K.F. (2020). The parkin coregulated gene product PACRG promotes TNF signaling by stabilizing LUBAC. Science signaling, 13(617), eaav1256.

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Quantifying p65 Nuclear Translocation

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Translocation of p65 from the cytosol to the nucleus was determined by indirect immunofluorescence based on single cell analysis. SH-SY5Y cells or MEFs were plated on glass coverslips and transfected with control, PACRG- or SHARPIN-specific siRNA. 24 h after transfection cells were treated with TNF (25 ng/ml) for 20 min and then fixed with 4% PFA in PBS for 10 min at room temperature. Cells were permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% donkey or goat serum in 0.2% Triton X-100 in PBS for 1 h at room temperature. Subsequently cells were incubated with an anti-p65 antibody diluted in blocking buffer overnight at 4°C. After extensive washing, cells were incubated with an ALEXA488-conjugated secondary antibody for 1 h at room temperature. Finally, the cells were washed extensively with PBS and then mounted onto glass slides and analyzed by fluorescence microscopy using a Nikon Eclipse E400 microscope. Nuclei were counterstained with DAPI (Sigma) and transfected cells were visualized by mCherry or mitoDsRed plasmid cotransfection. Cells were categorized in two classes: cells showing complete or incomplete nuclear translocation of p65.

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Golgi Staining of Hippocampal and Prefrontal Neurons

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Following sacrifice, brains were removed from subjects and cut into an anterior block (anterior to the optic chiasm) and a posterior block (between the optic chiasm and the brainstem) and placed in solutions provided in the Rapid Golgi Stain Kit (FD NeuroTechnologies, Ellicott City, MD). Golgi impregnation was performed as previously described (Frankfurt et al., 2011 (link); Inagaki et al., 2012 ). Secondary basal dendrites and tertiary apical dendrites were analyzed blindly from pyramidal cells in the CA1 region of the dorsal hippocampus and layer II/III of the prelimbic portion of the mPFC. Six cells per region/brain were included in the analysis and 5 or 6 brains were quantified per group. Neurons in both areas were chosen for analyses as follows: (1) cell bodies and dendrites were well impregnated, (2) dendrites were clearly distinguishable from adjacent cells and continuous. Spines were counted under oil (100x) using a hand counter and dendritic length measured using the Spot Advanced program, version 5.0 Windows (Diagnostic Instruments, Inc.) and a Nikon Eclipse E400 microscope. Spine density was calculated by dividing the number of spines by the length of the dendrite and data expressed as number of spines/ 10 μm dendrite.

Bowman R.E., Luine V., Khandaker H., Villafane J.J, & Frankfurt M. (2014). Adolescent Bisphenol-A exposure decreases dendritic spine density: Role of sex and age. Synapse (New York, N.Y.), 68(11), 498-507.

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Histological Analysis of CHIKV Infection

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WT mice were inoculated with 10³ FFU of CHIKV and treated with indicated anti-CHIKV or isotype control mAbs by intraperitoneal injection at 3 dpi. At 7 dpi, animals were perfused sequentially with PBS and 4% paraformaldehyde (PFA). Ipsilateral feet were collected, and hair was removed using Nair (Church & Dwight). Tissue was fixed for 24 h in 4% PFA, rinsed with PBS and water, and then decalcified for 10 days in 14% EDTA free acid (Sigma) in water at pH 7.2. Decalcified tissue was rinsed, dehydrated, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H & E).
Viral RNA ISH was performed on tissues from mice at 5 and 7 dpi. Tissue and slides were prepared as described above. ISH was performed using RNAscope 2.5 (Advanced Cell Diagnostics (ACD)) according to the manufacturer’s instructions and as previously described using the CHIKV probe (479501) designed and synthesized by ACD (54 ). Images were acquired on a Nikon Eclipse E400 microscope.

Fox J.M., Roy V., Gunn B.M., Huang L., Fong R.H., Edeling M.A., Mack M., Fremont D.H., Doranz B.J., Johnson S., Alter G, & Diamond M.S. (2019). Optimal therapeutic activity of monoclonal antibodies against chikungunya virus requires Fc-FcγR interaction on monocytes. Science immunology, 4(32), eaav5062.

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Histopathological Lung Assessment in Hamsters

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Animal handling, hamster infections, NTZ preparation and oral administrations were performed as described above. The anatomo-histological study was implemented as previously described.²⁴ (link)^,²⁶ Briefly, lungs were collected after intratracheal instillation of 4% (w/v) formaldehyde solution, and then fixed 72h at room temperature with a 4% (w/v) formaldehyde solution before being embedded in paraffin. Tissue sections of 3.5µm were stained with hematoxylin-eosin (H&E) and blindly analyzed by a certified veterinary pathologist. Microscopic examination was done using a Nikon Eclipse E400 microscope. Different anatomic compartments were examined (1) for bronchial and alveolar walls, a score of 0 to 4 was assigned based on severity of inflammation; (2) regarding alveoli, a score of 0 to 2 was assigned based on presence and severity of hemorrhagic necrosis; (3) regarding vessel changes (leucocytic accumulation in vascular wall or in endothelial compartment), absence or presence was scored 0 or 1 respectively. A cumulative score was then calculated and assigned to a grade of severity (see Supplementary Table 4).

Driouich J.S., Cochin M., Touret F., Petit P.R., Gilles M., Moureau G., Barthélémy K., Laprie C., Wattanakul T., Chotsiri P., Hoglund R.M., Tarning J., Fraisse L., Sjö P., Mowbray C.E., Escudié F., Scandale I., Chatelain E., de Lamballerie X., Solas C, & Nougairède A. (2022). Pre-clinical evaluation of antiviral activity of nitazoxanide against SARS-CoV-2. eBioMedicine, 82, 104148.

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Lung Histological Analysis Protocol

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Lung histological study was conducted as previously described (Driouich et al., 2021 (link)). Briefly, after collection, lungs were fixed with an intratracheal instillation of 4% (w/v) formaldehyde solution. Before being embedded in paraffin lungs were soaked during 72h at room temperature in a 4% (w/v) formaldehyde solution. Tissue sections of 3.5 μm, obtained following the “global open RENI” guidelines (standard reference in nomenclature and diagnostic criteria in toxicologic pathology https://www.goreni.org/), were stained with hematoxylin-eosin (H&E) and evaluated blindly by a certified veterinary pathologist. Microscopic observation was done using a Nikon Eclipse E400 microscope. The scoring system and the severity grade used in this study have been previously extensively described (Driouich et al., 2021 (link)).

Touret F., Driouich J.S., Cochin M., Petit P.R., Gilles M., Barthélémy K., Moureau G., Mahon F.X., Malvy D., Solas C., de Lamballerie X, & Nougairède A. (2021). Preclinical evaluation of Imatinib does not support its use as an antiviral drug against SARS-CoV-2. Antiviral Research, 193, 105137.

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In Vitro Fertilization of Oikopleura

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O. dioica were cultured as described [61]. For in vitro fertilization, mature female animals were transferred to watch glasses with 5 ml artificial seawater (Red Sea Salt 30 g/l, final salinity 30 PSU) and left to spawn. Sperm solutions were prepared from 3–4 male animals in 10 ml artificial seawater kept on ice. Solutions were checked for viability using a Nikon Eclipse E400 microscope and 50 µl aliquots were used to fertilize the oocytes spawned from one female. After 2.5 minutes, fertilized eggs were washed with artificial seawater. Embryos were left to develop at 18℃ to the desired stage.

Feng H., Raasholm M., Moosmann A., Campsteijn C, & Thompson E.M. (2019). Switching of INCENP paralogs controls transitions in mitotic chromosomal passenger complex functions. Cell Cycle, 18(17), 2006-2025.

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Evaluation of SPDL1 Expression

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The protein expression was evaluated at 20x original objective magnification in a blinded fashion by two independent researchers (DG, AKW), including one senior pathologist (DG) using the light ECLIPSE E400 microscope (Nikon Instruments Europe, Amsterdam, The Netherlands). All disagreements regarding SPDL1 staining were resolved by a consensus meeting. Immunoexpression of the examined protein was scored according to the modified Remmele-Stegner Index (IRS) being the product of the percentage of positively stained cells/areas (0–4) and staining intensity (0–3). The percentage of positive cells/area (PS) was evaluated as follows: (0) less than 10% of stained cells/area, (1) 11–20% of stained cells/area, (2) 21–50% of stained cells/area, (3) 51–80% of stained cell/area and (4) equal or more than 81% of stained cells/area. In turn, staining intensity (IS) was measured on a 4-point scale as (0) negative, (1) weak staining, (2) moderate staining, and (3) strong staining. To determine “high” or “low” expression levels of SPDL1, the final IRS scores were dichotomized based on the optimal cut-off point established with the cutp function of the Evaluate Cutpoints software [38 (link)]. The cut-point values for high and low expression groups were <7.5; ≥7.5, respectively.

Klimaszewska-Wiśniewska A., Buchholz K., Durślewicz J., Villodre E.S., Gagat M, & Grzanka D. (2022). SPDL1 Is an Independent Predictor of Patient Outcome in Colorectal Cancer. International Journal of Molecular Sciences, 23(3), 1819.

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Histological and Immunofluorescent Analysis of Murine Ear Inflammation

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Ears from mice sacrificed 4 days after DNFB (or OVA) challenge or re-challenge were excised and flash frozen in OCT compound. Skin cross-sections (7 μm thick) were stained with hematoxylin and eosin and imaged with a Nikon Eclipse E400 microscope. For fluorescent immunohistochemistry (IHC), 10 μm thick sections were fixed with 96% ethanol, blocked with PBS containing 5% donkey serum and 1% Tween20, and treated with a streptavidin/biotin blocking kit (Vector Labs, Burlingame, CA). Blocked sections were incubated overnight at 4 °C with primary antibodies: biotin-FoxP3 (FJK-16s; eBio), biotin-CD8a (53–6.7, eBio), or CD3 (SP7, monoclonal rabbit IgG; Thermo Scientific, Waltham, MA). Sections were then incubated with Cy3-streptavidin (Jackson ImmunoResearch Laboratories, West Grove, PA) or Alexa Fluor 555 donkey anti-rabbit IgG (Thermo Scientific) for 1 h at room temperature, counterstained with DAPI, and fixed with 2% paraformaldehyde. Slides were imaged with an epifluorescence microscope (Olympus Provis AX-70; Center Valley, PA). For histology and IHC, ears from naïve mice were used as controls.

Balmert S.C., Donahue C., Vu J.R., Erdos G., Falo LD J.r, & Little S.R. (2017). In vivo induction of regulatory T cells promotes allergen tolerance and suppresses allergic contact dermatitis. Journal of controlled release : official journal of the Controlled Release Society, 261, 223-233.

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