Quemesa

Manufactured by Thermo Fisher Scientific

The Quemesa is a laboratory equipment product designed for sample preparation and analysis. It provides a reliable and consistent method for sample heating, digestion, and evaporation. The core function of the Quemesa is to facilitate the preparation of samples for various analytical techniques.

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Lab products found in correlation

Fei tecnai spirit electron microscope, by Thermo Fisher Scientific (2 mentions) Tecnai spirit transmission electron microscope, by Thermo Fisher Scientific (1 mentions) Tecnai spirit electron microscope, by Thermo Fisher Scientific (1 mentions) Reichert ultracut s ultramicrotome, by Leica (1 mentions) Ab9110, by Abcam (1 mentions) A11122, by Thermo Fisher Scientific (1 mentions)

5 protocols using quemesa

Ultrastructural Analysis of Tregs

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After 24 h of rest, Tregs are seeded on concanavalin A–coated coverslips (Sigma) and incubated for 30 min at 37 °C. Cells are then fixed in 2,5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.4 for 1 h, post-fixed for 1 h with 2% buffered osmium tetroxide, dehydrated in a graded series of ethanol solution, and then embedded in epoxy resin. Images were acquired with a digital camera Quemesa (SIS) mounted on a Tecnai Spirit transmission electron microscope (FEI Company) operated at 80 kV.

Bonnin E., Rodrigo Riestra M., Marziali F., Mena Osuna R., Denizeau J., Maurin M., Saez J.J., Jouve M., Bonté P.E., Richer W., Nevo F., Lemoine S., Girard N., Lefevre M., Borcoman E., Vincent-Salomon A., Baulande S., Moreau H.D., Sedlik C., Hivroz C., Lennon-Duménil A.M., Tosello Boari J, & Piaggio E. (2024). CD74 supports accumulation and function of regulatory T cells in tumors. Nature Communications, 15, 3749.

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Correlative Light and Electron Microscopy

Cited 2 times

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CLEM was performed largely as described before (Verweij et al., 2018 (link)). In short, HeLa cells were seeded on sapphire discs coated with 20-nm carbon and finder grids, and transfected with CD63-pHluorin- and ORP1L-encoding plasmids 24 h later. The following day, sapphire discs were transferred to 3-mm carrier discs and high-pressure frozen using (HPM Live μ, CryoCapCell). After cryo-immobilization, the specimens were freeze substituted with Lowicryl (HM20) in an AFS-2 as previously described (Heiligenstein et al., 2014 (link)). The blocks were trimmed and sectioned in 250- and 90-nm sections, after which the sections were recovered on grids, labeled with Hoechst, and imaged on a wide-field microscope at different magnifications to facilitate correlation with EM data. Grids were contrasted and examined with a Tecnai Spirit electron microscope (FEI Company), and digital acquisitions were made with a numeric camera (Quemesa; Soft Imaging System). Correlations between light- and electron microscopy data were performed using eC-CLEM (Paul-Gilloteaux et al., 2017 (link)).

Verweij F.J., Bebelman M.P., George A.E., Couty M., Bécot A., Palmulli R., Heiligenstein X., Sirés-Campos J., Raposo G., Pegtel D.M, & van Niel G. (2022). ER membrane contact sites support endosomal small GTPase conversion for exosome secretion. The Journal of Cell Biology, 221(12), e202112032.

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Visualizing Exosome Dynamics in Zebrafish

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To induce expression of CD63-pHluorin, specifically labeling exosomes, embryos were injected at the 1000 cell stage. The following day, embryos were anesthetized with tricaine and embedded in 1.5% low melting point agarose. Live imaging recordings were performed at 28°C using a Nikon TSi spinning-wide (Yokagawa CSU-W1) microscope (Verweij et al., 2019 (link)). Other embryos were used for ultrathin cryosectioning and immunogold labeling, in which case they were fixed in 2% PFA, 0.2% glutaraldehyde in 0.1M phosphate buffer pH 7.4 at 3 dpf. Zebrafish were processed for ultracryomicrotomy and immunogold labeled against GFP using PAG 10. All 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) (Verweij et al., 2019 (link)).

Thouvenin O., Keiser L., Cantaut-Belarif Y., Carbo-Tano M., Verweij F., Jurisch-Yaksi N., Bardet P.L., van Niel G., Gallaire F, & Wyart C. (2020). Origin and role of the cerebrospinal fluid bidirectional flow in the central canal. eLife, 9, e47699.

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Ultrastructural Analysis of Melanocytes

Cited 1 time

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Control and Rab4A-knockdown melanocytes were seeded on Matrigel coated glass coverslips. Post 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 (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 (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. Melanosomes and vacuoles per µm2 cytosol were counted using ImageJ software. We have counted 10 cells each from control sh and Rab4A sh condition. Further, we have estimated the melanosome stages from 883 total melanosomes of control sh and 300 total melanosomes of Rab4A sh cells.

Nag S., Rani S., Mahanty S., Bissig C., Arora P., Azevedo C., Saiardi A., van der Sluijs P., Delevoye C., van Niel G., Raposo G, & Setty S.R. (2018). Rab4A organizes endosomal domains for sorting cargo to lysosome-related organelles. Journal of Cell Science, 131(18), jcs216226.

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Ultrastructural Analysis of HCMV-US28 and iHA-US28

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Doxycyclin-induced iHA-US28-U251 cells and HCMV-US28-pHluorin-infected HFFF-Tet cells were fixed in 2% PFA, 0.2% glutaraldehyde in 0.1M phosphate buffer pH 7.4. Cells were then washed with phosphate buffer, embedded in 10% (wt/vol) gelatin and infused in 2.3 M sucrose. Mounted gelatin blocks were frozen in liquid nitrogen and ultrathin sections were prepared with an Ultracut FCS ultracryomicrotome (Leica). Ultrathin cryosections were labeled with rabbit anti-HA (cat. #: ab9110, Abcam, 1:500) or rabbit anti-GFP (cat. #: A11122, Invitrogen, 1:200) antibodies, and protein A coupled to 10nm gold particles. EVs and virions were isolated from the culture medium of HCMV-infected HFFF-Tet cells. Culture medium was centrifuged at 500xg to remove dead cells and cell debris and subsequently centrifuged for 1 hour at 23,000 rpm to isolate EVs and virions. EVs and virions were spotted on carbon-coated and formvar coated EM grids and fixed with 2% PFA in PBS before staining with anti-GFP (antibody and protein A coupled to 10nm gold particles. 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).

Bebelman M.P., Setiawan I.M., Bergkamp N.D., van Senten J.R., Crudden C., Bebelman J.P., Verweij F.J., van Niel G., Siderius M., Pegtel D.M, & Smit M.J. (2023). Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging. iScience, 26(8), 107412.

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