Immunocytochemistry of cytokeratin 19 (CK19), α-smooth muscle actin (α-SMA), vimentin (VIM) and fibroblast activation protein (FAP) was performed to verify the purity of OVCAFs and OVNFs. Briefly, cells on sterile glass coverslips were fixed in ice-cold methanol and then incubated overnight at 4°C in a humid chamber with the indicated primary antibodies as follow: 1:200 mouse anti-human CK-19 antibody (sc-6278, Santa Cruz Biotechnology Inc.), 1:500 anti-α-SMA antibody (A5228, Abcam), 1:500 anti-VIM antibody (sc-6260, Santa Cruz Biotechnology Inc.), and 1:500 rabbit anti-human FAP (ab53066, Abcam). After washing out the excess primary antibody, the coverslip was incubated for 3 h at room temperature with the appropriate secondary fluorescent antibody including goat anti-mouse IgG-Cy3 antibody (1:2,000, #115-166-071, Jackson ImmunoResearch Laboratories Inc.) or the goat anti-rabbit IgG-FITC antibody (1:2,000, ab6717, Abcam). Hoechst 33342 solution (Invitrogen; Thermo Fisher Scientific, Inc.) was added to stain the nucleus. Fluorescence was captured using an Inverted microscope model IX71 (Olympus Corporation).
Inverted microscope model ix71
Manufactured by Olympus
Sourced in Germany
The Olympus Inverted Microscope (model IX71) is a versatile instrument designed for observing and analyzing samples from the underside. It features a compact, ergonomic design and advanced optical components to provide high-quality images for a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg cy3 antibody, by Jackson ImmunoResearch (1 mentions)
Hoechst 33342 solution, by Thermo Fisher Scientific (1 mentions)
Sc 6260, by Santa Cruz Biotechnology (1 mentions)
Ab53066, by Abcam (1 mentions)
A5228, by Abcam (1 mentions)
Sc 6278, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit igg fitc antibody, by Abcam (1 mentions)
μ slide 1 luer, by Ibidi (1 mentions)
4 protocols using inverted microscope model ix71
1
Immunophenotyping of Ovarian Cancer-Associated Fibroblasts
2
In Vitro Magnetic Retention Assay Under Flow
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In vitro magnetic retention assays were carried out under flow conditions in a modified channel slide (µ-Slide I Luer, 0.4-mm height, ibidi) using a two-magnet system as previously described [38 (link)]; the neodymium iron boron (NdFeB) permanent magnets (Supermagnete) used in this study are shown in Table 2 . Briefly, OT-I CD8+ T cells (107 cells/ml) were treated with MNPs (150 µg Fe/ml) or not for 2 h in standard conditions, and then washed and stained. An Olympus Inverted Microscope (model IX71) coupled to an Imaging Station cell^R under standard culture conditions was used in this assay. Shear stress was set at 0.5 dyne/cm2 and events were recorded every 1 s. After 60 s, the two-magnet system was applied for the next 2 min. Imaris software (Bitplane) was used to analyse displacement along the Y-axes (in the direction of the magnetic field).
3
Inhibition of HAdV-GFP Infection in A549 Cells
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HAdV-GFP (37.5 ng) was incubated with compound 16 (50 µM) or the same volume of DMSO for 1 h in complete DMEM. Parallel samples were incubated for 10 min at the indicated temperatures, cooled to r.t., and added to A549 cells on ice to synchronize the infection. After the incubation, cells were washed twice with PBS 1x to remove the nonattached virus and the excess of compound 16 and DMSO. Cells were then incubated for 24 h at 37 °C and 5% CO2 before being visualized for GFP expression in an Olympus inverted microscope Model IX71 (Hamburg, Germany) and analyzed by capturing representative images of each condition using the CellSens Dimension platform (Olympus, Hamburg, Germany).
4
In vitro magnetic cell retention
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In vitro magnetic retention assays were carried out under flow conditions in a modified channel slide (μ-Slide I Luer, 0.4-mm height, ibidi) using a two-magnet system as previously described (43 (link)). The neodymium iron boron (NdFeB) permanent magnets (Supermagnete) used in this study were the following: an 5 × 14-mm magnet with 1.35 T of remanent magnetization (Br) (magnet A) and an 8 × 6-mm magnet with 1.45 T of remanent magnetization (Br) (magnet B). Briefly, either murine NK cells expanded in vitro in the presence of IL-2 or the human NK-92MI cell line (107 cells/ml) were treated with MNPs (150 μg Fe/ml) or not for 2 h in standard conditions, and then washed and stained with calcein-AM. An Olympus Inverted Microscope (model IX71) coupled to a Cell∧R imaging station under standard culture conditions was used in this assay. Shear stress was fixed at 0.5 dyne/cm2 and events were recorded every 1 s. After 60 s, the two-magnet system was applied for the next 2 min. Imaris software (Bitplane) was used to analyse displacement along the Y-axes (in the direction of the magnetic field).
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