2.3. Single-cell suspension preparation The preparation of single-cell suspension was followed by our previous protocols [15,20]. Briefly, testes tissues from the Ctrl and triptolide-treated mice were digested with collagenase IA and deoxyribonuclease I (Thermo Fisher Scientific Inc., Waltham, MA,...
However, dissociation of tumor tissue into a single cell suspension prior to orthotopic transplantation enables the injection of fewer cell numbers, the selection of tumor-initiating populations by specific purification using antibody staining and fluorescence-activated cell sorting, and the analysis of ...
To determine cell types, we filtered differentially expressed genes between target cluster and other cells using FindAllMarkers function in Seurat (Pvalue ≤ 0.05, log2FC ≥ 0.585). The cells were annotated with cell-type-specific expression known in the literature. Moreover, SingleR and...
Precancerous lesions of the oral mucosa, especially those accompanied by moderate to severe dysplasia, contribute to the initiation of oral squamous cell carcinoma (OSCC). However, the cellular compositions and spatial organization of the precancerous st
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 for 2 min). Both kidneys were harvested and the...
Preparation of single-cell suspension Each tissue sample was transferred to a sterile 10-cm culture dish and cut into < 1 mm3 segments before being transferred to a 50-ml centrifuge tube. Each tissue type was then digested in the DMEM (Sigma-Aldrich, USA) containing 100 μg/ml collagenase ...
Specifically, cell encapsulation was the first step of the workflow. Cell suspension was injected into the chip and dispersed with a relatively slow speed (0.5 μl/min) (Fig. 2a(i)) into the continuous oil phase (6 μl/min) at the “T-junction” part of the channels (Fig. 2a...
(Additional file2: Fig. S2B). Single cell suspension was then visualized under a microscope, and the cell viability was examined. The results showed that we successfully obtained a sufficient number of single cells with over 90% viability (Additional file2: Fig. S2C, D), and copies of JEV...
cell lin RESULTS Evaluation of cell loss during loading of DEPArray cartridges The DEPArray cartridges are typically loaded with 14 μl of a fluorescently labelled single-cell suspension. To prevent the injection of air in the main microchamber of the cartridge where the visualisation and sorting...
The proteomic processing steps start by introducing a cell suspension (500 or 25 cells/μL) into cell capture chambers under flow pressure of 3 psi, with which a quantifiable number of cells in the range of 1, 5, 10, 50, and 100 were trapped in a cell trapping chamber by controlli...