E600 upright microscope

Manufactured by Nikon

Sourced in United States

The Nikon E600 upright microscope is a high-performance laboratory instrument designed for various microscopy applications. It features a stable and ergonomic design, allowing for precise and reliable observations. The E600 provides a wide range of magnification options and optical capabilities to support a variety of research and analysis needs.

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Lab products found in correlation

Micromax, by Teledyne (2 mentions) Metamorph imaging software, by Universal Imaging (2 mentions) Control igg, by BioXCell (1 mentions) Diff quik, by Merck Group (1 mentions) Anti ifn γ clone xmg1, by BioXCell (1 mentions) Permount, by Merck Group (1 mentions) Neutrophil elastase, by Abcam (1 mentions) Bond refine polymer staining kit, by Leica (1 mentions) Bond max automated staining system, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Tcs sp5 confocal microscope, by Leica (1 mentions) Cd163, by Abcam (1 mentions) Image pro, by Media Cybernetics (1 mentions) Formaldehyde, by Merck Group (1 mentions) Ki 67 clone d3b5, by Cell Signaling Technology (1 mentions) Ds ri1 digital camera, by Nikon (1 mentions) Abc kit, by Vector Laboratories (1 mentions) Nis elements software, by Nikon (1 mentions) Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions)

11 protocols using e600 upright microscope

Culturing and Infecting Protozoan Parasites

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RH, GT1, Prugniaud (Pru), VEG strain T. gondii; and Nc1 [75] (link), Nc2 [76] (link) and NcLiv strain N. caninum were maintained by serial passage in human foreskin fibroblast (HFF) monolayers as described previously [77] (link). RH stably expressing mCherry were obtained from Dr. Anita Koshy [78] (link). Primary bovine fibroblasts were provided by Dr. Kenneth John McLaughlin (Penn). Cryopreserved bone marrow cells recovered from Myd88^−/−, Myd88/Trif^−/−, and Tlr2/Tlr4^−/− mice were differentiated into macrophages as described previously [79] (link), and infected with a 1∶1 MOI of parasites for QPCR and microarray experiments at 16–22 hours post-infection. Ifnar1^−/− and Tlr3^−/− mice were provided by Drs. Hao Shen and Yongwon Choi (Penn), respectively. For mouse infections, WT and Ifnar1^−/− mice received either control IgG or anti-IFN-γ (clone XMG-1.2, 1mg I.P., BioXCell) on day 0 and again at 4 days post-infection. Mice were infected with 3×10⁶ NcLiv strain Neospora caninum by I.P. injection on day 0. All mice were maintained at the University of Pennsylvania in accordance with Institutional Animal Care and Use Committee guidelines. For cytospins, 200,000 cells were spun onto glass slides using a Shandon Cytospin and stained with Diff-Quik, dehydrated, mounted with Permount (Sigma Aldrich), and images captured on a Nikon E600 upright microscope.

Beiting D.P., Peixoto L., Akopyants N.S., Beverley S.M., Wherry E.J., Christian D.A., Hunter C.A., Brodsky I.E, & Roos D.S. (2014). Differential Induction of TLR3-Dependent Innate Immune Signaling by Closely Related Parasite Species. PLoS ONE, 9(2), e88398.

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Multiplex Immunofluorescence Analysis of Tumor Immune Landscape

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Following deparaffinization and rehydration, heat induced antigen retrieval was performed using Tris-EDTA buffer pH 9. Tissues were stained for S100A9 (Novus Biologicals), CD33 (Novocastra), CD8, Neutrophil Elastase, and CD163 (Abcam). PD-1 antibody was obtained from R&D Systems. The following fluorescently conjugated secondary mAb (Life technologies) were used: anti-rabbit IgG AF594 for S100A9 and Neutrophil Elastase, anti-mouse IgG AF647 for CD33 and anti-mouse IgG AF594 for CD163. Alexa-594 donkey anti-mouse IgG was used for CD8 and Alexa-647 donkey anti-goat antibody for PD1. Cell nuclei were stained using DAPI (Life technologies). Images were obtained using Leica TCS SP5 Confocal microscope. 16 frames at 63X magnification were used to calculate the cell count/mm2.
IHC Staining was performed on a Bond Max automated staining system (Leica Microsystems). The Bond Refine polymer staining kit (Leica Microsystems) was used. FoxP3 mAb (Biolegend), CD4 mAb (Leica microsystems) and CD8 mAb (DAKO) were used and antigen retrieval was performed with ER2 and ER1 (Leica Microsystems) retrieval solutions. Slides were rinsed, dehydrated through a series of ascending concentrations of ethanol and xylene and then mounted. Images were obtained using Nikon E600 Upright Microscope. 12 frames at 40X magnification were used to calculate the cell count/mm².

Dominguez G.A., Condamine T., Mony S., Hashimoto A., Wang F., Liu Q., Forero A., Bendell J., Witt R., Hockstein N., Kumar P, & Gabrilovich D.I. (2016). Selective targeting of myeloid-derived suppressor cells in cancer patients using DS-8273a, an agonistic TRAIL-R2 antibody. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(12), 2942-2950.

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Quantitative Analysis of Immune Cell Populations

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BAL cells were collected from naïve mice of naïve and
infected/co-infected mice (3 days after PR8 infection) and cytospins performed using a
Cytospin3 centrifuge (Thermo-Shandon). Cells were fixed and stained using the Kwik-Diff
stain kit (Thermo-Shandon) as described (Iparraguirre et
al., 2008). Images were captured with a Nikon E600 upright microscope (original
magnification: 60×) and processed with the image software Image Pro (Media
Cybernetics) or Nikon Elements D software.

Wolf A.I., Strauman M.C., Mozdzanowska K., Williams K.L., Osborne L.C., Shen H., Liu Q., Garlick D., Artis D., Hensley S.E., Caton A.J., Weiser J.N, & Erikson J. (2014). Pneumolysin Expression by Streptococcus pneumoniae Protects Colonized Mice from Influenza Virus-induced Disease. Virology, 0, 254-265.

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Microscopic Imaging of Gummy Candies

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Gummy candies samples were cut into blocks of 2 mm thick using a razor blade. The internal structures of each sample were exposed by cutting the gummy in halves. The samples were mounted on a slide. Reflected light microscopy images of samples were recorded with Nikon E600 upright microscope (Nikon Instruments, Melville, NY, USA) equipped with a bright 100 W halogen illumination and a black and white Lumera Infinity 3-1UC camera with 1.4 Mpx for image recording. Objective lens was Nikon 10×/0.25 Ph1 and working distance 200 mm. Illumination was set manually while the gain of the camera was adjusted automatically. Images were recorded using Infinity Capture version 5.0.4 program.

Otálora M.C., Coralia O.s.o.r.i.o., Helber de Jesús B.a.r.b.o.s.a., Jairo Ernesto P.e.r.i.l.l.a, & Mónica Azucena N.a.z.a.r.e.n.o. (2019). Encapsulated betalains (Opuntia ficus-indica) as natural colorants. Case study: Gummy candies. Lebensmittel-Wissenschaft + [i.e. und] Technologie. Food science + technology. Science + technologie alimentaire, 103.

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Nanoparticle Biodistribution in Mice

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Whole organs (brain, liver, kidney, and spleen) of C57BL/6 mice were removed through necropsy 120 h after intravenous injection of nanoparticles or PBS and preserved in 10% formalin for 48 h. Tissues were then embedded in paraffin wax, sliced into 5 μm thick sections, and stained with hematoxylin and eosin (H&E) or Prussian blue/Nuclear Fast Red using standard clinical laboratory protocols. Microscopic images of tissues were acquired using an E600 upright microscope (Nikon) equipped with a CCD color camera. Blood cell panels and serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were quantified 120 h after intravenous administration of nanoparticles or free drug (n = 3 per treatment condition), and compared to mice receiving PBS injection (n = 5). Blood (300 μL) was drawn from each mouse through cardiac heart puncture. Samples were then submitted to a veterinary pathology laboratory (Phoenix Laboratories, Everett, WA) for third party analysis.

Stephen Z.R., Kievit F.M., Veiseh O., Chiarelli P.A., Fang C., Wang K., Hatzinger S.J., Ellenbogen R.G., Silber J.R, & Zhang M. (2014). Redox-Responsive Magnetic Nanoparticle for Targeted Convection-Enhanced Delivery of O6-Benzylguanine to Brain Tumors. ACS Nano, 8(10), 10383-10395.

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Telomere Visualization in Fibroblast Cells

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We treated fibroblast cells transfected with WT and mutant telomerase with 100 μg/ml of colcemid for 7 hrs. The cells were grown to 70% confluence after 21 passages on a 10 cm plate. Cells were then trypsinized to detach them from the plate, pelleted and treated in a hypertonic environment (75 mM KCl for 30 mins at 37° C) to rupture them. We fixed the cells in 10 ml of 3:1 methanol:acetic acid solution and stored at 4° C. Metaphase spreads were fixed in 4% formaldehyde (Sigma), treated with 1 mg/ml pepsin in 10 mM glycine, pH 2.0 at 37° C (Sigma) and fixed again with 4% formaldehyde. The slides were then dehydrated with ethanol, air dried, and hybridized with 20 μL of 200 nM telomeric-Cy5 peptide nucleic acid (PNA) probe (TelC-Cy5 - PNA biosciences) (LiCor) according to the manufacturer’s instructions. Slides were counterstained with DAPI and imaged using a Nikon E600 upright microscope.

Bryan C., Rice C., Hoffman H., Harkisheimer M., Sweeny M, & Skordalakes E. (2015). Structural Basis of Telomerase Inhibition by the Highly Specific BIBR1532. Structure (London, England : 1993), 23(10), 1934-1942.

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Microscopic Quantification of Microvessels

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Under microscopy, localization of labeled microvessels was performed with a Nikon E600 upright microscope with a ×20 objective lens. The microscope was coupled to cooled charge‐coupled device camera (Micromax; Princeton Instruments Inc, Trenton, NJ). Five nearby 1‐mm² images were taken from each of 3 sections in the frontal cortex of each brain or in the GSN‐Ms, and the mean MVD within these images was taken to represent cortical or skeletal muscle MVD in that animal. All acquired images from individual sections were analyzed for number of microvessels using MetaMorph Imaging software (Universal Imaging Co, Downingtown, PA) or Nikon Elements software.41

Frisbee S.J., Singh S.S., Jackson D.N., Lemaster K.A., Milde S.A., Shoemaker J.K, & Frisbee J.C. (2018). Beneficial Pleiotropic Antidepressive Effects of Cardiovascular Disease Risk Factor Interventions in the Metabolic Syndrome. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 7(7), e008185.

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Quantitative Analysis of Ki67 Expression

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Tissues were embedded in Tissue-Tek OCT and frozen. Endogenous peroxidases were quenched from acetone-fixed sections (8 μm) by incubating in 0.3% H₂O₂ for 10 minutes at room temperature. Following quenching, sections were blocked using 3% goat serum followed by staining with antibodies against Ki-67 (clone D3B5, Cell Signaling Technology). Immunohistochemistry using the ABC Kit (Vector labs) was performed according to the manufacturer’s instructions, and sections were counter stained with hematoxylin. Slides were then imaged at 10X objective magnifications on a Nikon E600 Upright microscope with a Nikon DS-Ri1 Digital camera. Nikon NIS-Elements software was used for image acquisition and image stitching of the entire tumor. Image Pro Plus 7 analysis software was used to measure the percentage of Ki67 stained nuclei within each sample. Percentage of stained nuclei was calculated as total area of brown stained nuclei divided by total area of sample.

Allegrezza M.J., Rutkowski M.R., Stephen T.L., Svoronos N., Perales-Puchalt A., Nguyen J.M., Payne K.K., Singhal S., Eruslanov E.B., Tchou J, & Conejo-Garcia J.R. (2016). TRAMETINIB DRIVES T CELL-DEPENDENT CONTROL OF K-RAS-MUTATED TUMORS BY INHIBITING PATHOLOGICAL MYELOPOIESIS. Cancer research, 76(21), 6253-6265.

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Collagen-based Cell Invasion Assay

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250 × 10³ cells were added to collagen-based cell invasion chambers (#ECM551, Millipore, MA, USA) and incubated for 24, 48 or, 72 h (three separate experiments, each in duplicate) at 37 °C in 5% CO₂. FBS (10%) was used as a chemo attractant. Solutions containing the NPs (100 nM of SP, 100 nM CGRP, 50 nM of NKA or their combination) or vehicle (media) were replaced every 24 h. After the treatment was completed, noninvasive cells from the interior of the insert were removed. Inserts containing invasive cells were stained, washed, imaged (using a 10× objective and a Nikon E600 upright microscope equipped with a CCD digital camera), and lysed with the extraction buffer provided in the collagen-based invasion assay (50% of Reagent Alcohol [90% ethanol, 5% methanol and 5% isopropanol] in 50 mM acetic acid, pH 4.5). Dye mixture was transferred to a 96-well plate and absorbance at 560 nm was measured using a spectrophotometer (Epoch Microplate Spectrophotometer, BioTek Instruments Inc, Vermont, USA).

Gutierrez S, & Boada M.D. (2018). Neuropeptide-induced modulation of carcinogenesis in a metastatic breast cancer cell line (MDA-MB-231LUC+). Cancer Cell International, 18, 216.

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Histological Analysis of Nanoparticle-Treated Brains

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Whole brains were removed through necropsy 48 h post-CED of NPs and preserved in 10% formalin for 48 h. Whole brains were then embedded in paraffin wax, sliced in to 5 μm thick sections, and stained with hematoxylin and eosin (H&E) or Prussian blue/Nuclear fast red using standard clinical laboratory protocols. Microscopic images of brains were acquired using an E600 upright microscope (Nikon) equipped with a CCD color camera. Additional preserved brains were provided to the University of Washington Pathology Research Services Laboratory to acquire electron micrographs.

Stephen Z.R., Chiarelli P., Revia R.A., Wang K., Kievit F., Dayringer C., Jeon M., Ellenbogen R, & Zhang M. (2019). Time-resolved MRI assessment of convection-enhanced delivery by targeted and non-targeted nanoparticles in a human glioblastoma mouse model. Cancer research, 79(18), 4776-4786.

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