How can I seed 0.5x10^6 cells per well in a 96-well plate (I only need to seed 5 wells)?
I have a T25 flask (5 mL total volume) with a cell density of 0.6x10^6 cells/mL.
To seed 0.5x10^6 cells per well in a 96-well plate (but only 5 wells) and considering you have a T25 flask with a cell density of 0.6x10^6 cells/mL, you can follow these steps:
1. Calculate Total Cells Needed:
0.5×10^6 cells per well x 5 wells = 2.5×10^6 total cells needed.
2. Calculate Volume of Cells Needed:
(2.5×10^6 cells): (0.6 ×10^6 cells/mL)=4.17mL
This is the total volume of the cell suspension needed to get the required number of cells.
3. Prepare Cell Suspension:
Take the cell suspension, centrifugate at 2000 rpm for 5 minutes and remove the supernatant of the existing media (to make space for the cell suspension).
4. Add Fresh Media:
Add 4,17 mL of fresh media to the cells pellet and gently mix the cells to ensure a homogeneous cell suspension.
6. Transfer to 96-Well Plate:
Dispense 0.83 mL (830 μL) of the cell suspension into each of the 5 wells in the 96-well plate.
7. Incubation:
Place the 96-well plate in the incubator for cell attachment and growth.
Ensure that you maintain proper sterile techniques throughout the process to avoid contamination. Additionally, be mindful of the timing and conditions required for the specific cell type you are working with.
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