16HBE cells were seeded in glass coverslips and exposed to LPS or to the TRPV4 agonist GSK1016790A during 5 min. When indicated, cells were pre-incubated for 15 min with the TRPV4 inhibitor HC067047 (10 µM). After treatment, cells were fixed with cold paraformaldehyde and permeabilized with 0.2% Triton X-100. Primary antibody against iNOS (1:1000; PA3-030A, ThermoFisher Scientific) was incubated overnight at 4 °C, followed by anti-rabbit Alexa Fluor Plus 488 (1:1000; A32731, Invitrogen) for 1 h at room temperature. Coverslips were mounted in glass slides using DAPI-containing mounting solution (VectaShield, Vector Laboratories, Burlingame, CA, USA). The confocal images of labeled cells were collected using the optimal pinhole size for the ×63 oil objective of a Zeiss LSM 510 Meta Multiphoton microscope (Carl Zeiss AG, Oberkochen, Germany). We used a custom-designed software in Matlab (The MathWorks, Inc) to determine the number and size of iNOS aggresomes.
Lsm 510 meta multiphoton microscope
Manufactured by Zeiss
Sourced in Germany
The Zeiss LSM 510 Meta Multiphoton microscope is a laser scanning confocal microscope designed for high-resolution fluorescence imaging. It utilizes multiphoton excitation to enable deep tissue imaging with reduced phototoxicity. The microscope is equipped with multiple laser lines and high-sensitivity detectors to capture detailed optical sections of biological samples.
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Lab products found in correlation
2 protocols using lsm 510 meta multiphoton microscope
1
Quantifying iNOS Aggresomes in 16HBE Cells
2
Biaxial Tissue Mechanical Loading Device
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To provide real-time monitoring of biaxial loads applied over a thin tissue segment mounted on the stage of a high-resolution nonlinear microscope, a biaxial mechanical loading device (Fig. 1A) was developed as described in detail in our previous publication (5) . In a few words, this lightweight device consists of four loading grips connected to adapters for clamping the tissue, one stage insert, one platform, four pulleys, four tension screws, and four force gauges. The entire device was placed on the motorized X-Y stage on a Zeiss LSM 510 Meta Multiphoton microscope (Carl Zeiss Microscopy). The force extended on each grip was monitored in real time using four digital force gauges with 50 mN resolution (Mecmesin, West Sussex, UK). In a near-frictionless situation, small pulleys guide the thin cables from grips to the corresponding force gauges. To achieve tension control, small grips held the tissue specimen at all four sides of the tissue segments. Tension control was achieved by placing a 10 -32 screw between each grip and its corresponding force gauge in a way that screw rotation increased tension in the cable. The force reading was calibrated by applying an initial pretension with a typical value of 0.15 N.
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