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Quemesa

Manufactured by Olympus
Sourced in Germany, Japan, Netherlands

The Quemesa is a high-performance laboratory equipment designed for scientific applications. It serves as a versatile instrument capable of performing various analytical tasks. The core function of the Quemesa is to provide reliable and accurate measurements, enabling researchers and scientists to conduct their experiments and analyses with precision.

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35 protocols using quemesa

1

Ultrastructural Analysis of Melanocyte Organelles

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Control and Rab4A-knockdown melanocytes were seeded on Matrigel-coated glass coverslips. After 24 h, cells were fixed initially with 0.5% Karnovsky's fixative (4% paraformaldehyde, 72 mM sodium cacodylate pH 7.4, 4 mM CaCl2, 0.5% glutaraldehyde) for 2 h followed by overnight fixation with 2% Karnovsky's fixative (contains 2% glutaraldehyde). Cells were processed for Epon embedding as described previously (Raposo et al., 2001 (link)). Ultrathin sections of cell monolayers were prepared with a Reichert UltracutS ultramicrotome (Leica Microsystems) and contrasted with uranyl acetate and lead citrate as described previously (Raposo et al., 2001 (link)). Samples were examined with a FEI Tecnai Spirit electron microscope (FEI Company), and digital acquisitions were made with a numeric camera (Quemesa; Soft Imaging System). For quantification, melanosome stages were defined by morphology (Raposo et al., 2001 (link)) and vacuoles were defined as empty organelles. The number of melanosomes and vacuoles per µm2 cytosol were counted using ImageJ software. We counted ten cells from each control sh and Rab4A sh condition. Furthermore, we estimated the melanosome stages from 883 total melanosomes of control sh and 300 total melanosomes of Rab4A sh cells.
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2

TEM Imaging of Hydrophilic Gel Samples

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Transmission electron microscopy (TEM) micrographs were acquired on a Philips EM 208 equipped with Quemesa (Olympus Soft Imaging Solutions) camera; images were acquired with RADIUS software; samples were prepared as follows. Carbon-copper grids were first exposed to the UV-ozone cleaner (Procleaner Plus) for 15 min to make the grid surface more hydrophilic, then gels were precisely deposited on a TEM grid, dried for 15 min at RT, and contrasted by aqueous tungsten phosphate solution (pH 7.4).
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3

Transmission Electron Microscopy Imaging

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TEM micrographs were acquired using a Philips electron microscope 208 (FEI, Hillsboro, Oregon, OR, USA) that was equipped with a Quemesa (Olympus Soft Imaging Solutions (Berlin, Germany) camera; images were recorded with RADIUS software; samples were prepared as described previously using phosphotungstate as a negative stain [24 (link)].
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4

Liposomal Vesicles Characterization via TEM

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Morphological and structural characterization of uncoated and chitosan-coated liposomal vesicles was done using transmission electron microscopy (TEM) employing an EM 208 (Philips) system equipped with a camera (Quemesa, Olympus Soft Imaging Solutions). For this analysis, samples were diluted 1 : 1 with distilled water, then deposited on a Formvar/carbon support film on a specimen grid (Electron Microscopy Sciences). After air-drying for 5 min, the sample was negatively stained with 1% (w/v) of uranyl acetate solution for 10 min.
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5

Characterization of Ferrous Sulfate-Loaded Nanoliposomes

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Morphological characterizations of unloaded and ferrous sulfate–loaded nanoliposomes were performed by transmission electron microscopy (TEM) (EM 208, Philips, Amsterdam, Netherlands) equipped with camera Quemesa (Olympus Soft Imaging Solutions, Münster, Germany). The samples were negatively stained with 1% (w/v) of uranyl acetate solution.
Fe-nanoliposomes were also observed by the optical microscope in fluorescence field Axioplan 2- Image Zeiss, Jena, Germany, equipped with software to capture the images. The Rhodamine B dye was used to visualize lipid vesicles with a 100 X oil immersion objective.
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6

Transmission Electron Microscopy of Small Extracellular Vesicles

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Procedures were performed essentially as described (Raposo et al., 1996 (link); Hurbain et al., 2017 (link)).
For transmission electron microscopy (TEM), sEV preparations were loaded on copper formvar/carbon coated grids (Ted Pella). Fixation was performed with 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4), followed by a second fixation with PBS 1% glutaraldehyde in PBS. Samples were stained with 4% uranyl acetate in methylcellulose.
For immunolabeling electron microscopy (IEM), sEV preparations were loaded on grids and fixed with 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). Immunodetection was performed with a mouse anti-human CD63 primary antibody (Abcam ab23792). Secondary incubation was next performed with a rabbit anti mouse Fc fragment (Dako Agilent Z0412). Grids were incubated with Protein A-Gold 10 nm (Cell Microscopy Center, Department of Cell Biology, Utrecht University). A second fixation step with 1% glutaraldehyde in PBS was performed. Grids were stained with uranyl acetate in methylcellulose.
All samples were examined with a Tecnai Spirit electron microscope (FEI, Eindhoven, The Netherlands), and digital acquisitions were made with a numeric 4k CCD camera (Quemesa, Olympus, Münster, Germany). Images were analyzed with iTEM software (EMSIS) and statistical studies were done with Prism-GraphPad Prism software (v8).
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7

Immunoelectron Microscopy of Fixed Cells

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Cells were fixed with a mixture of 2% (w/v) paraformaldehyde and 0.2% (w/v) glutaraldehyde in 0.1 M PHEM buffer (120 mM PIPES, 50 mM HEPES, 4 mM MgCl2, 20 mM EGTA pH 6.9), and processed for immunoelectron microscopy as described previously (Setty et al., 2007 (link)). Ultrathin cryosections were prepared using UC7 ultracryomicrotome (Leica, Vienna, Austria) (Raposo et al., 1997 ), double immunogold labeled with protein A conjugated to 10- or 15-nm gold particles (PAG10, PAG15 from Cell Microscopy Center, AZU, Utrecht, The Netherlands) and analyzed under a Tecnai Spirit electron microscope (FEI, Eindoven, The Netherlands) equipped with a 4k CCD camera (Quemesa, Olympus).
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8

Immunodetection of Extracellular Vesicles

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Procedures were performed essentially as described [25 (link),32 (link),33 (link)]. Immunodetection was carried out with the following primary antibodies: mouse anti-human CD63 (Abcam ab23792), mouse anti-human CD9, or mouse anti-human CD81 (both from Dr E. Rubinstein, Université Paris-Sud, Institut André Lwoff, Villejuif, France). Secondary incubation was performed with a rabbit anti mouse Fc fragment (Dako Agilent Z0412), then grids were incubated with Protein A-Gold 10 nm (Cell Microscopy Center, Department of Cell Biology, Utrecht University). All samples were observed with a Tecnai Spirit electron microscope (FEI, Eindhoven, The Netherlands), and digital acquisitions were made with a numeric 4k CCD camera (Quemesa, Olympus, Münster, Germany). Images were analysed with iTEM software (EMSIS) and statistical studies were done with GraphPad Prism software (v8).
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9

Phage Visualization by Electron Microscopy

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Pf4* phages were resuspended in PBS and a drop of suspension was placed on a petri dish. A Formvar EM grid was placed Formvar side down on top of the phages drop for approximately 1 min. The grid was removed, blotted with filter paper, and placed onto a drop of 7.0% uranyl acetate in distilled water for 3 min. The grid was washed in water, blotted again with filter paper, and examined on EM JEOL 1010 at 80 KV. The pictures were recorded by using Quemesa, the electron digital camera of Olympus.
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10

Immunolabeling and TEM Imaging of HTT Aggregates

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Total brain homogenate was centrifuged at 18,000 x g at 4 C for 20 min; the resulting supernatant was pelleted by ultra-centrifugation at 190,000 x g for 40 min and resuspended in 10 mM Tris-HCl (pH 7.4). Immunolabeling was performed with minor modifications as described (Laue, 2010) . Briefly, samples were incubated on formvar-coated copper grids (Plano) for 10 min before immunolabeling. Grids were blocked and washed in PBS supplemented with 1% BSA and 0.1% glycine. Labeling was performed with the anti-HTT aggregate antibody Agg53 and an appropriate 12 nm colloidal gold-labeled secondary antibody (Jackson ImmunoResearch). Samples were stained with 2% uranyl acetate and imaged with a Zeiss EM 910 transmission electron microscope at 80 kV. Acquisition was performed with a CDD camera (Quemesa, Olympus Viewing System).
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