The level of intracellular calcium ions of spermatozoa was determined by flow cytometry (FCM). The acetoxymethyl ester form of fluo-4 (fluo-4/AM, Keygen, Nanjing, China) was used as a fluorescent calcium ion indicator. Spermatozoa were incubated for 15 min at room temperature with 5 μmol l−1 fluo-4/AM and 0.001% Pluronic F-127 (Keygen) in a sperm nutrition solution. The cells were then washed and incubated with the same solution for 30 min. Then, flow cytometry was used to measure and capture the intensity of images of calcium-containing sperm cells.
Pluronic f 127
Manufactured by Keygen Biotech
Sourced in United States
Pluronic F-127 is a non-ionic triblock copolymer composed of polyethylene glycol and polypropylene glycol. It is a widely used surfactant and thermoreversible gel-forming agent in various pharmaceutical and biomedical applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pluronic f 127
1
Intracellular Calcium Levels in Sperm
2
Calcium Imaging in Rat Chondrocytes
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Rat chondrocytes were loaded with 5 μM fluo-3-acetoxymethyl ester (Fluo-3-AM, Sigma, USA) and Pluronic F-127 (KeyGEN BioTECH, China) containing 25 mM HEPES (Sigma, USA) for 30 min at 37 °C in the dark, then washed twice with PBS to remove extracellular Fluo-3-AM. Imaging was performed using Leica inverted microscope and analyzed with ImageJ software. The measured average fluorescence intensity of each cell in the field (F) was normalized with the non-specific background fluorescence (F0) to obtain the fluorescence intensity (F/F0).
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