Gm 4000 co2 supplier

For short-term imaging experiments, images were acquired with an IX81 inverted microscope (Olympus, Tokyo, Japan), which was equipped with a Retiga 4000R cooled Mono CCD camera (QImaging, Surrey, Canada), a Spectra-X light engine illumination system (Lumencor, Inc., Beaverton, OR, USA), an IX2-ZDC laser-based autofocusing system (Olympus), a UPlanSApo 60x/1.35 oil objective lens (Olympus), a MAC5000 controller for filter wheels and XY stage (Ludl Electronic Products, Hawthorne, OR, USA), an incubation chamber (Tokai Hit, Fujinomiya, Japan), and a GM-4000 CO₂ supplier (Tokai Hit). The following filters and dichroic mirrors were used: for CFP, an FF01-438/24 excitation filter (Semrock, Lake Forest, IL, USA), an XF2040 445DRLP dichroic mirror (Omega, Biel, Switzerland), and an FF01-483/32-25 emission filter (Semrock); for FRET, an FF01-438/24 excitation filter (Semrock), an XF2040 445DRLP dichroic mirror (Omega), and an FF01-542/27 emission filter (Semrock). The microscopes were controlled using MetaMorph software ver. 7.8.11.0 (Molecular Devices, San Jose, CA, USA). The images were acquired every 20 s for 10 min. An amount of 100 uM H₂O₂ (Wako) or H₂O was added at 2 min, followed by the addition of 1 mM DTT or H₂O at 6 min.

CFP and YFP images of HeLa cells and A549 cells were obtained by using an inverted microscope (IX81-ZDC; Olympus, Tokyo, Japan) equipped with a cooled CCD camera (Cool SNAP-K4; Roper Scientific), an illumination system (Spectra-X light engine; Lumencore, OR), an IX2-ZDC2 laser-based autofocusing system (Olympus), a MAC5000 controller for filter wheels and XY stage (Ludl Electronic Products, Hawthorne, NY), an incubator chamber system (Tokai Hit, Shizuoka, Japan) and a GM-4000 CO2 supplier (Tokai-Hit, Fujinomiya, Japan). The following filters were used for the dual emission imaging studies: FF01–438/24–25 (Semrock, Rochester, NY, USA), and FF01–475/28–25 (Semrock) excitation filter for CFP and YFP/GFP, a U-MREF glass reflector (Olympus) as a dichroic mirror, an FF01–483/32–25 emission filter (Semrock) for CFP and an FF01–542/27–25 emission filter (Semrock) for YFP. The microscope was controlled by MetaMorph software (Universal Imaging, West Chester, PA). The average fluorescence intensities of CFP and YFP in each cell were measured by manually delineating a region of interest at the cytoplasm with MetaMorph software.