Product Description
EnkiLife Lymphocyte Separation Medium (LSM) is a ready-to-use separation medium that has been filtersterilized. It is suitable for the in vitro isolation of human peripheral blood lymphocytes and can also be used to isolate lymphocytes from other tissues, including umbilical cord blood and bone marrow cells. Each 100 mL of LSM contains 6.2g of polysucrose and 9.4g of sodium diatrizoate. The density of LSM is 1.0770-1.0800 g/mL at 20℃.
Principle
The density of red blood cells and granulocytes in human blood is approximately 1.090 g/mL. Lymphocytes and
monocytes have a density of 1.075–1.090 g/mL, while platelets have a density of 1.030–1.035 g/mL. By using
defibrinated or heparin-treated blood, diluted 1:1 with normal saline or balanced salt solution, and performing
low-speed centrifugation for 30 minutes using a density gradient solution with a density between 1.075 and
1.092 (nearly isotonic), cells of specific densities will distribute according to the density gradient, thereby
separating different types of blood cells.
Red blood cells and polymorphonuclear granulocytes migrate through the gradient and settle at the bottom of
the tube. Lymphocytes and other mononuclear cells (monocytes and platelets) are distributed between the
plasma and the LSM layers. Lymphocytes can be recovered by aspirating the interphase solution and then
further washed to remove platelets, LSM, and plasma.
Product Specifications
Transport and Storage
| Parameter | Specification | Important Notes |
|---|---|---|
| Transport | Room temperature | - |
| Storage | 15-25°C, protected from light | DO NOT refrigerate or freeze. Storage at 4°C may cause white crystallization, compromising separation performance. |
| Handling | All operations must be performed under sterile conditions. | - |
Optimal Separation Time
| Time Post-Collection | Separation Performance |
|---|---|
| Within 2 hours | Optimal |
| 2-4 hours | Acceptable |
| 4-6 hours | Decreased viability, poor separation |
| Over 6 hours | Very poor separation, may fail completely |
Critical Requirement
Optimal Separation Conditions
- Aseptic Technique: Strictly follow aseptic procedures in a laminar flow hood or biosafety cabinet.
- Temperature: Perform procedure at 18-22°C ambient temperature; 20°C yields optimal results. Temperature deviations may alter separation medium density and compromise performance.
- Equilibration: Allow reagents to equilibrate to recommended temperature before use (20°C in summer, 25°C in winter).
Precautions
Use appropriate consumables: Avoid high-polymer materials (e.g., polystyrene) due to static electricity. Use static-free or low-static centrifuge tubes and non-alkali-treated glassware, as static causes cell adhesion to tube walls and alkali-treated glass surfaces become roughened, compromising separation efficiency.
Avoid plasma contamination: Aspirating excessive material above the target cell layer will introduce plasma proteins and platelets.
Proper dilution is critical: Improper dilution reduces cell yield and viability. Diluent requirements：Calcium/magnesium-free buffer or medium. If blood is diluted, reduce centrifugal force and time accordingly.
