LLO-mediated permeabilized HeLa cells were incubated with resealing buffer (1 mM ATP, 8 mM creatine kinase and 50 μg/ml creatine phosphate, 1 mg/ml glucose, 1 mM GTP, 25 mM Hepes-KOH, 115 mM potassium acetate, 25 mM MgCl2, pH7.4) that contained the molecules indicated in the text at 37 °C for 30 min. Then, 1 mM CaCl2 was added and the cells were incubated at 37 °C for 5 min. The cells were washed twice with medium and were further incubated with pre-warmed medium at 37 °C, 5% CO2 for more than 30 min. The cells were washed twice with PBS, and were observed by using an LSM510 confocal microscope or were subjected to flow cytometry using Guava easyCyte HT flow cytometry system (Millipore).
Guava easycyte ht flow cytometry system
Manufactured by Merck Group
Sourced in United States
The Guava easyCyte HT flow cytometry system is a compact and automated instrument designed for cell analysis. It utilizes flow cytometry technology to rapidly measure and characterize individual cells within a sample. The system is capable of detecting and quantifying various cellular parameters, including size, granularity, and the expression of specific markers.
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Lab products found in correlation
3 protocols using guava easycyte ht flow cytometry system
1
Permeabilization and Resealing of HeLa Cells
2
Annexin V-APC Apoptosis Assay
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At the fifth day after lentivirus transfection, A549 cells were stained with 200 μl cell suspension containing 10 μl Annexin V-APC (Cat. 88-8007, eBioscience, USA) at room temperature in the dark for 10–15 min, then flow cytometry analysis was performed on the Guava easyCyte HT flow cytometry system (Millipore).
3
Cell Cycle Analysis by Flow Cytometry
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Cell cycle distribution was analyzed with flow cytometry. The logarithmically growing A549 cells were trypsinized and resuspended in DMEM with 10% FBS, then rinsed with pre-cold D-Hanks. After centrifugation, the supernatant was removed and cells were fixed in 70% cold ethanol for 1 h. Fixed cells were rinsed with D-Hanks solution and permeabilized with 0.1% Triton X-100 and 2 mg/ml RNase A in D-Hanks solution for 30 min. The cells were then rinsed with D-Hanks solution and stained with 50 mg/ml propidium iodide (PI) (Sigma-Aldrich, USA). Stained cells were analyzed with the Guava easyCyte HT flow cytometry system (Millipore, USA).
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