Eyes were harvested from fish not used in the experiments to measure retinal responses to light stimuli. Food was withheld for 24 hours and 10–12 fish from each diet were anesthetized in a bath containing 25 mg L-1 MS-222 (Syndel, Ferndale, WA, USA) buffered with 50 mg L-1 sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA). Upon removal from the anesthesia bath, the spinal cord was immediately severed, and eyes harvested. Retinal tissue was subsequently embedded, sectioned, mounted on glass slides, and stained with Hematoxylin-Eosin using standard histological procedures. Images were captured using a BX53 microscope outfitted with a DP73 camera and visualized with the cellsSens software (version 510, Olympus, Center Valley, PA, USA).
L 1 sodium bicarbonate
Manufactured by Merck Group
Sourced in United States
L-1 sodium bicarbonate is a laboratory chemical used as a pH regulator. It is a white, crystalline powder that is soluble in water. The primary function of L-1 sodium bicarbonate is to maintain a desired pH level in various laboratory applications.
Automatically generated - may contain errors
2 protocols using l 1 sodium bicarbonate
1
Harvesting and Histological Analysis of Fish Retinas
2
Deuterium Labeling of HEK293T Cells
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Human embryonic kidney (HEK293T) cells (UC Berkeley cell culture facility) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with L-glutamine and without pyruvate, including 10% fetal bovine serum (FBS), and 1% penicillin/ streptomycin. Cells were passaged upon achieving between 70-100% confluency. Cells were imaged at 50-70% confluency. Cells were transferred to and imaged in the live cell imaging solution (Fisher Sci).
To deuterium tag cells, the D 2 O (Sigma) DMEM media was made by mixing 70% D 2 O (Sigma) with 30% deionized H 2 O to dissolve DMEM powder (Sigma), 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 3.7 g L -1 sodium bicarbonate (Sigma). After pH adjusting to 7.4, the media was sterilized with a 0.2 μm filter. The cell media was replaced with the D 2 O media at 50-70% cell confluency then incubated for 20-24 hours. The cells were transferred to and imaged in the live cell imaging solution with 70% D 2 O.
To deuterium tag cells, the D 2 O (Sigma) DMEM media was made by mixing 70% D 2 O (Sigma) with 30% deionized H 2 O to dissolve DMEM powder (Sigma), 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 3.7 g L -1 sodium bicarbonate (Sigma). After pH adjusting to 7.4, the media was sterilized with a 0.2 μm filter. The cell media was replaced with the D 2 O media at 50-70% cell confluency then incubated for 20-24 hours. The cells were transferred to and imaged in the live cell imaging solution with 70% D 2 O.
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