The imaging buffer used was 10 mM tris containing 100 mM cysteamine hydrochloride (Sigma-Aldrich, M6500), glucose oxidase (4.0 mg/ml; Sigma-Aldrich, G0543), catalase (0.57 mg/ml; Sigma-Aldrich, C40-100MG), and 10% glucose (Sigma-Aldrich, 49158-1KG-F). pH was adjusted with 0.5 M NaOH to pH 8.3 to 8.5. Chamber wells were completely filled with buffer and sealed bubble free (air free) by a regular coverslip. Images were recorded using an inverted total internal reflection fluorescence (TIRF) microscope (Olympus IX-71) equipped with an oil-immersion objective lens (Olympus, UApoN, ×100 magnification, NA 1.49, TIRF) and a 647-nm laser (PhoxX 647, 140 mW, Omicron Laserage, Germany). A quad-edge dichroic beam splitter (Di01-R405/488/561/635, Semrock) and a quad-band excitation filter (FF01-446/523/600/ 677, Semrock) were used to remove the laser light before imaging the fluorescence on an EMCCD camera cooled to −50°C (DU-885-CS0-#VP, Andor) with an effective pixel size of 80 nm. Raw movies typically contained 3000 to 4000 images recorded at 30 to 90 Hz. Images were analyzed using rapidSTORM and custom-written routines in Matlab.
C40 100mg
Manufactured by Merck Group
The C40-100MG is a laboratory equipment product. It is a compact and precise device designed for specialized laboratory applications. The core function of this product is to provide controlled and consistent measurements, without further interpretation or extrapolation on its intended use.
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Lab products found in correlation
2 protocols using c40 100mg
1
Single-Molecule TIRF Imaging Protocol
2
Oxygen Scavenger Buffer Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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In this research, we used oxygen scavenger buffer (Glox-buffer). We prepared a glucose stock solution (300 mM glucose, 50 mM Tris, 10 mM NaCl dissolved in Milli-Q H2O) and stored it at 4°. The final concentration of each ingredients are 1.25 mg/ml catalase (Sigma, C40-100MG), 1 mg/ml glucose-oxidase (Sigma, G2133-10KU), and 50~150 mM MEA (Sigma, 30070-10G) diluted in glucose stock. We adjusted the blinking density by adjusting the concentration of MEA.
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