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Calculations II

Learn how to calculate volumes of stock solutions based on molarity and feeding concentrations for cell cultures. This guide provides two approaches to convert feeding concentrations into molarity, using example exercises with compounds X and Y. It emphasizes high accuracy considerations when the added volume is significant. Additionally, it details cell suspension calculations for preparing wells in a 24-well plate, ensuring optimal cell density. Master these calculations to effectively manage your laboratory cultures.

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Calculations II

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  1. Calculations II

  2. Working With Molarity (M) • In Some Cases Molarity (M) Is Known. • Ex. 10-5 M Compound X (MW 325) • You Also Know That Cultures Should Be “fed” 30 ng/mL Of Compound X • What Volume Of Compound X Stock Solution Should You Use To “feed” 10 mL Of Culture?

  3. Working With Molarity (M) One Approach Is To Convert 30 ng/mL of cpd X Into Molarity

  4. Working With Molarity (M) • Alternative Approach

  5. Exercise • Compound Y: 3x10-5 M, (MW 425), “Feed” 5 mL Culture with 10 ng/mL • Ans: 3.9 L

  6. When High Accuracy Is Needed • If volume added is significant Ex. 1 mL, and Culture Volume is 10 mL • First determine V1 Based On Culture Volume (V2=10 mL) • Re-determine V1 Based on New Culture Volume (10mL+V1+0.5mL extra)

  7. When High Accuracy Is Needed

  8. Exercise • Determine what volume to use from stock vial • Culture volume is 11 ml • Stock vial concentration is 1mM • “Feeding” concentration is 0.1 mM • Final Culture Volume Should Be Between 12.25-12.5 mL

  9. Solution

  10. Cell Calculations • Cell Suspension of 2 mL @ 5x106 cells/mL • Use a 24 well plate, Prepare 12 wells @ 0.25x106 cells/mL, 1 mL in each well • Add 50 L To Each Well • Add 950 L Medium

  11. Cell Calculations • Cell Suspension of 10 mL @ 0.25x106 cells/mL • Use a 25cm2 flask, Prepare 5 mL @ 1x106 cells/mL • You will have to resuspend cells in smaller volume • Even then, not enough cells for 5 mL @ 1x106 cells/mL • Enough for 2.5 mL @ 1x106 cells/mL

  12. Exercises • Prepare 12 wells @ 0.25x106 cells/mL • Well volume: 0.5 mL • Cell Suspension concentration: 10x106 cells/mL, 2 mL • Answer: 13 L+487 L • How Many wells can you prepare? • Answer: (20x106/0.125x106) =160 wells

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