Briefly, the specimens were digested at 37 °C for 15 min and passed through 40-micron sterile strainer (Corning). Cells were centrifuged at 300 × g for 5 min and cell pellets were resuspended in 1 ml PBS (HyClone). Cell suspensions were counted to determine cell viability, and the concentration was adjusted to 1 × 105 cells/ml. Single-cell suspension was then loaded to microfluidic chip (GEXSCOPE Single Cell RNA-seq Kit, Singleron Biotechnologies) and scRNA-seq libraries were constructed according to the manufacturer’s instructions (Singleron Biotechnologies). Libraries were sequenced on Illumina Novaseq 6000 platform with 150 bp paired end reads.
40 micron sterile strainer
Manufactured by Corning
Sourced in United States
The 40-micron sterile strainer is a laboratory filtration device designed to remove particles and impurities from liquid samples. It features a 40-micron pore size filter mesh that captures particles while allowing the desired liquid to pass through. The strainer is manufactured in a sterile environment to maintain sample integrity.
Automatically generated - may contain errors
Lab products found in correlation
Tc20 automated cell counter, by Bio-Rad (3 mentions)
Gexscope tissue dissociation solution, by Singleron Biotechnologies (3 mentions)
Gexscope tissue preservation solution, by Singleron Biotechnologies (2 mentions)
Gexscope red blood cell lysis buffer, by Singleron Biotechnologies (2 mentions)
Gexscope single cell rna seq kit, by Singleron Biotechnologies (1 mentions)
Novaseq 6000 platform, by Illumina (1 mentions)
4 protocols using 40 micron sterile strainer
1
Single-cell RNA-seq of Dissociated Cells
2
Single-Cell Tissue Dissociation Protocol
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Fresh samples were stored in the GEXSCOPE Tissue Preservation Solution (Singleron Biotechnologies, Nanjing, China) at 2–8 °C immediately after being collected by needle biopsy or bronchoscopy. Prior to dissociation, tissue samples were washed with Hanks Balanced Salt Solution (HBSS) for three times, minced into small pieces, and digested in 2 ml GEXSCOPE Tissue Dissociation Solution (Singleron Biotechnologies) following manufacturer’s instructions. Briefly, the specimens were digested at 37 °C for 15 min with continuous agitation. A 40-micron sterile strainer (Corning) was used to separate cells from impurities after digestion. The cells were then centrifuged at 300 × g 4 °C for 5 min and cell pellets were resuspended in 1 ml PBS (HyClone). Cell suspensions were counted with TC20 automated cell counter (Bio-Rad) to determine cell concentration and viability.
3
Efficient Tissue Dissociation and Cell Isolation
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Prior to tissue dissociation, the specimens were washed with Hanks balanced salt solution (HBSS) three times and minced into 1–2 mm pieces. The tissue pieces were digested in 2 mL GEXSCOPETM Tissue Dissociation Solution (Singleron Biotechnologies) at 37 °C for 15 min in a 15 mL centrifuge tube with continuous agitation. Following digestion, a 40-micron sterile strainer (Corning) was used to separate cells from cell debris and other impurities. The cells were centrifuged at 1000 rpm and 4 °C for 5 min, and the cell pellet was resuspended in 1 mL PBS (HyClone). To remove red blood cells, 2 mL GEXSCOPETM Red Blood Cell Lysis Buffer (Singleron Biotechnologies) was added to the cell suspension and incubated at 25 °C for 10 min. The mixture was then centrifuged at 1000 rpm for 5 min, and the cell pellet was resuspended in PBS. Cells were counted with a TC20 automated cell counter (Bio-Rad).
4
Single-cell isolation from kidney tissue
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Fresh kidney tissue was immediately transferred into GEXSCOPE® Tissue Preservation Solution (Singleron Biotechnologies, Nanjing, Jiangsu Province, China) at 2°C to 8°C. The samples were digested in 2 mL of GEXSCOPE® tissue dissociation solution (Singleron Biotechnologies) at 37°C for 15 min in a 15-mL centrifuge tube with continuous agitation after being washed with Hanks balanced salt solution three times and cut into approximately 1 to 2 mm pieces. Subsequently, cell debris and other impurities were filtered by a 40-micron sterile strainer (Corning, NY, USA). The cells were centrifuged at 300 × g and 4°C for 5 min. Cell pellets were resuspended in 1 mL of PBS (HyClone, Marlborough, MA, USA). To remove red blood cells, 2 mL GEXSCOPE® Red Blood Cell Lysis Buffer (Singleron Biotechnologies) was added to the cell suspension and incubated at 25°C for 10 min. The mixture was then centrifuged at 500 × g for 5 min, and the cell pellet was resuspended in PBS. Cells were counted with a TC20 automated cell counter (Bio-Rad, Hercules, CA, USA).
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