Single-cell suspension preparation Bladder tumors and adjacent normal mucosa were processed immediately after being obtained from bladder cancer patients. Every sample was cut into small pieces (<1 mm in diameter) and then was incubated with 2 ml of trypsin (Gibco, Cat: R001100), 1 ml of ...
Cell preparation A total of 4 different frozen human PBMCs samples were analyzed in this experiment. Frozen vials containing cells were thawed for 2 min in water bath at 37°C. After this, cell suspension was transferred to a fresh 2 ml Eppendorf tube using wide bore pipette tip (The...
The process for preparing a single-cell suspension is largely determined by the origin of the cells. Lauren Jachimowicz, application development scientist, flow cytometry at Agilent Technologies, notes that cultured cell lines can readily be transformed into a single-cell suspension eit...
Single-cell suspension preparation and scRNA-seq library construction Peripheral blood was collected from the donor into a heparin anticoagulation tube. After adding 10 mL of 6% hydroxyethyl solution, the sample was inverted several times and kept at room temperature for 30 min. Subsequently, the sup...
Cell suspension concentrations were kept low (300 cells/ul), to avoid macrophage aggregation. The cell suspension was counted with the Cellometer K2 (Nexcelom Bioscience, Lawrence, MA, USA) using AO/PI staining solution. The cell viability was > 90% in all samples. The microfluidic ...
(130-109-398) during the preparation of the single-cell suspension for scRNA-seq. In order to effectively isolate neural cells and remove cell fragments from brain tissues, the debris removal reagent from Miltenyi was employed for single-cell isolation prior to RNA sequencing and flow cytometry....
that are more challenging to dissociate into single cell suspension as opposed to infiltrating immune cells28. Therefore, even though immune cell infiltration is a common characteristic of ccRCC, the exact cellular composition of tumors in our as well as other scRNA-seq datasets16,18,20is likely ...
Single-cell/nucleus suspension and library preparation Single-cell suspension Glomeruli were enriched by depleting the PT segment from the kidneys using Percoll density gradient centrifugation described previously [13]. Briefly, the mouse was euthanized by cervical dislocation under anesthesia (4% isoflurane...
Single-cell RNA-sequencing library preparation The concentration of single-cell suspension was adjusted to 1 × 105cells/mL in PBS. Single-cell suspension was then loaded onto a microfluidic chip (GEXSCOPE Single Cell RNA-seq Kit, Singleron Biotechnologies) and scRNA-seq libraries were construc...
scRNA sequencing library preparation The concentration of the single-cell suspension was adjusted to 1 × 105 cells/mL in PBS. A single-cell suspension was then loaded onto a microfluidic chip75 (part of the Singleron GEXSCOPETM Single Cell RNA-seq Kit, Singleron Biotechnologies), and singl...