Cell fractionation

• Why is cell fractionation important in biology?

  It isolates organelles for study
• What are the steps involved in cell fractionation?

  1. Homogenization: Breaking open cells
  2. Ultracentrifugation: Separating organelles by density
• What type of solution is required for cell fractionation?

  Cold, isotonic, and buffered solution
• Why must the solution be cold during cell fractionation?

  To reduce enzyme activity that can damage organelles
• What does isotonic mean in the context of cell fractionation?

  Water potential is the same as organelles
• Why is it important for the solution to be isotonic?

  To prevent osmosis affecting organelles
• What is the consequence of not using a buffered solution?

  Sudden pH changes could damage organelles
• What is the process of homogenization in cell fractionation?

  • Cells are broken open using a blender
  • The solution must be cold, isotonic, and buffered
• What is the purpose of filtering the solution after homogenization?

  To remove large cell debris
• What happens during ultracentrifugation?

  Organelles are separated based on density
• What is the role of the centrifuge in cell fractionation?

  It spins samples at high speeds
• What is differential centrifugation?

  Separating organelles at increasing speeds
• How does the density of organelles affect their separation during centrifugation?

  More dense organelles move to the bottom first
• Which organelle is separated first during centrifugation?

  Nuclei
• What is the order of organelle separation during centrifugation?

  Nuclei, chloroplasts, lysosomes, ribosomes
• What is the significance of cell fractionation in studying organelles?

  • Isolates organelles for detailed study
  • Enables examination of structure and function
• What is the final step after isolating organelles?

  Examine the organelles
• What are the key requirements for successful cell fractionation?

  • Cold, isotonic, buffered solution
  • Proper homogenization and centrifugation techniques