To prepare a solution from 10 g of Ficoll Genei™ powder:

1. Weigh 10 g of Ficoll powder accurately.

2. Add the powder to a suitable container.

3. Add distilled water gradually while stirring.

4. Continue adding water until the desired final volume is reached (e.g., 100 mL for a 10% w/v solution).

5. Stir thoroughly until the powder is completely dissolved.

6. Sterilize the solution if needed (e.g., by autoclaving or filtration).

7. Store the solution properly (usually at room temperature or 4°C, depending on instructions).

can PBS be used in Ficoll solution preparation? if water is used ,it will cause the cells to swell and burst . i`m not sure of the exact reason, but i think water should not be used.

Hello Vennela Marla

For separating PBMCs from whole blood using Ficoll solution, you may prepare it in a buffer (like PBS or HBSS) and not directly in water. This is because the buffer maintains the proper osmotic balance necessary for cell viability during the separation process. Using water can cause cells to lyse due to osmotic shock. Buffer like PBS, are designed to maintain a stable osmotic environment, preventing drastic changes in cell volume and preserving cell integrity during the separation procedure.

Protocol:

1. You may weigh the Ficoll powder, and slowly add PBS solution while continuously stirring to help the Ficoll dissolve.

2. Continue adding PBS, ensuring thorough mixing, until the desired final volume is reached (e.g., 100 mL for a 10% w/v solution).

3. Continue stirring until the Ficoll powder is completely dissolved and the solution is clear. Gentle heating may be required to aid dissolution.

4. If the Ficoll solution must be used in sterile applications, filter sterilization (e.g., using a 0.22-μm filter) or autoclaving may be required.

5. You may store the Ficoll solution at an appropriate temperature for your application (typically at room temperature or 4°C, depending on the instructions or purpose). But please note that Ficoll solution should be at room temperature when separating PBMCs from whole blood because cold Ficoll can cause PBMCs to clump or experience cold shock, impacting the separation efficiency. It is recommended to bring Ficoll solution to room temperature for 1-2 hours before layering it with blood.

Best.