Gexscope red blood cell lysis buffer rclb

The livers obtained from fibrotic liver of S. japonicum-induced mouse and healthy liver of normal mouse. The fresh tissues were stored in the sCelLive™ Tissue Preservation Solution (Singleron) on ice as quickly as possible. After washed with Hanks Balanced Salt Solution (HBSS) for three times, the specimens were minced into small pieces. To dissociated into single-cell suspensions, the specimens then digested with 3 mL sCelLive™ Tissue Dissociation Solution (Singleron) by Singleron PythoN™ Tissue Dissociation System at 37°C for 15 min. After filtered through a 40-micron sterile strainer, the cell was added the GEXSCOPE^® red blood cell lysis buffer (RCLB, Singleron), which was incubated at room temperature for 5-8 min to remove red blood cells. Centrifugation was performed at 300 × g 4°C for 5 mins to remove supernatant and single cells were resuspended softly in PBS. Finally, the cell viability was evaluated microscopically through staining with Trypan Blue.

Within 30 min of procurement from both LN and normal donors, the renal tissue samples were preserved in the sCelLiveTM Tissue Preservation Solution (Singleron, China) on ice. The samples were then rinsed three times with Hanks Balanced Salt Solution, cut into small pieces, and digested using 3 mL of sCelLiveTM Tissue Dissociation Solution with the Singleron PythoN Tissue Dissociation System at 37°C for 15 min. The cell mixture was gathered and passed through a 40‐micron sterile filter to separate the cells. After that, the GEXSCOPE red blood cell lysis buffer (RCLB, Singleron) was added to the mixture in a 1:2 (volume) ratio with the cells and incubated at room temperature for 5–8 min to eliminate red blood cells. The mixture was then centrifuged at 300xg and 4°C for 5 min to remove the supernatant and resuspended softly in phosphate buffered saline (PBS) (HyClone, USA).

After sample extraction (Figures 1C, D), the fresh cells samples were immediately stored in the sCelLiVE® Tissue Preservation Solution (Singleron) on ice. The sample of ascites were transferred to a 15-ml centrifuge tube. The samples were then filtered with 40 µm sterile strainers, and centrifuged at 1,000 rpm at 4°C for 5 min. Next, 2 ml GEXSCOPE® red blood cell lysis buffer (RCLB, Singleron) was added to lyse the red blood cells for 10 min. Finally, the single cell suspension was collected after re-suspension with PBS, and trypan blue (Sigma) staining was used to calculate cell activity and cell count under a microscope (7 (link)).