Volume and mass were analyzed with live imaging as previously described76 (link). Briefly, cells were trypsinized, counted, and resuspended in medium supplemented with 1 mg/ml Alexa Fluor 488 Dextran (MW 10 kD; Life Technologies) at a concentration of 106 cells/ml before being injected in PDMS chips coated with fibronectin with a known height of 23 µm. Finally, the chamber was immersed in medium to prevent evaporation. Acquisitions were performed on a Ti inverted (Nikon) or an Axio Observer microscope (Carl Zeiss) at 37 °C, with 5% CO2 atmosphere, using a ×20 dry objective (NA 0.5 phase), or a SFluor 20× objective (NA 0.75 without phase ring) for the mass measurements. Images were acquired using MetaMorph (Molecular Devices). Images were acquired with a CoolSNAP HQ2 camera (Photometrics) for fluorescence or a PHA SICS camera (PHA SICS) for mass measurements. Images were analyzed with a custom-made MatLab program (MathWorks).
Ti inverted
Manufactured by Nikon
The Ti inverted is a research-grade inverted microscope designed for laboratory applications. It features a compact and ergonomic design, providing a stable platform for various imaging techniques. The core function of the Ti inverted is to enable high-quality observation and analysis of specimens in a wide range of life science and material science research fields.
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Lab products found in correlation
Axio observer microscope, by Zeiss (2 mentions)
Metamorph, by Molecular Devices (2 mentions)
Alexa fluor 488 dextran, by Thermo Fisher Scientific (1 mentions)
Matlab program, by MathWorks (1 mentions)
Intensilight, by Nikon (1 mentions)
Spectra x light, by Lumencor (1 mentions)
Axiovision, by Zeiss (1 mentions)
2 protocols using ti inverted
1
Live Cell Volume and Mass Quantification
2
Fluorescence and Mass Microscopy Protocol
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Acquisitions were performed on a Ti inverted (Nikon) or an Axio Observer microscope (Carl Zeiss) at 37°C, with 5% CO2 atmosphere, using a 10× dry objective (NA 0.30 phase), a 20× dry objective (NA 0.5 phase), or a SFluor 20× objective (NA 0.75 without phase ring) for the mass measurements. Images were acquired using MetaMorph (Molecular Devices) or Axio Vision (Carl Zeiss) software. The excitation source was either a mercury lamp (Intensilight; Nikon) or a LED illumination (SPECTRA X light; Lumencor or Zeiss Colibri). Images were acquired with a CoolSNAP HQ2 camera (Photometrics) for fluorescence or a PHASICS camera (PHASICS) for mass measurements.
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