Inquiry10X genomics single cell sequencing, animal cell culture, flow cytometry analysis of cells experiments must be based on single cells. Different methods of digesting tissues and dispersing cells can be chosen according to the characteristics of different solid tissue components in order to achieve high single-cell yield and low cell damage.
CD Genomics is committed to finding technical ways to treat different tissues with as little cell damage as possible. Our single cell suspension preparation method has a high cell yield and maintains the original characteristics of the cells.
10x Single Cell Suspension Preparation workflow
Sample requirements
For the preparation of solid tumor single cell suspensions, the site of tumor sampling is very important. Because there are degeneration or necrosis areas in larger tumor tissues, the use of degenerated tissues should be avoided as much as possible, and the sites with better vitality should be selected.
Cancer samples are placed in sample preservation solution. Seal and mark the centrifuge tubes.
The time between the removal of cancer tissue and its placement in the sample preservation solution is as short as possible and should be less than 1 hour. After its placement into the preservation solution, it is transported and stored in a refrigerated environment (i.e., 4-10°C).
In order to prevent the contamination of experimental materials, aseptic operation must be taken when collecting materials.
FAQ
What are the common tissue types that CD Genomics can digest?
Such as mouse spleen, liver, heart, lung, brain tissue, tumor tissue, human tumor tissue, nerve, skin tissue, etc.
If the sample is adherent cells cultured in vitro, does it still need to be digested?
Yes, generally trypsin is most commonly used at a concentration of 0.25%-5% and a digestion time of 1-5min at 37°C.
What should be the size of the tumor tissue sample?
The shape and area of the cancerous tissues need to be considered comprehensively, and the necrotic parts and non-cancerous normal tissues should not be mixed in.
For tissues with a large number of tumor cells, the size should be about 1 cm3 (0.5-1.0 g); for tissues with more interstitium, the size should be about 3 cm3 (1.0-3.0 g); for tiny samples, the effective cancerous tissue should be ensured to be greater than 0.3 g.
What is the most appropriate concentration range for the cell storage solution before cell counting?
An important factor that affects the accuracy of cell counting is the concentration of the cell storage solution. It has been found that a concentration of cell storage solution in the range of 700-1200 cells/µl is optimal. Exceeding this optimal range may result in unreliable results. If the sample is not within this range, it is recommended to adjust the concentration of the cell storage solution accordingly.
