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Cell Fractionation by Centrifugation Video Lecture | Crash Course for CSIR NET Life Sciences

FAQs on Cell Fractionation by Centrifugation Video Lecture - Crash Course for CSIR NET Life Sciences

1. What is cell fractionation, and why is it important in cell biology?

Ans.Cell fractionation is a laboratory technique used to separate cellular components while preserving their individual functions. This process is crucial in cell biology as it allows researchers to isolate specific organelles, proteins, and other cellular structures for analysis. Understanding the composition and functions of these components helps in studying cellular processes, disease mechanisms, and developing medical therapies.

2. What is differential centrifugation, and how does it work?

Ans.Differential centrifugation is a method of cell fractionation that involves spinning a cell suspension at various speeds. This process separates cellular components based on their size and density. Initially, the suspension is centrifuged at low speeds to sediment larger organelles, such as nuclei. The supernatant is then collected and subjected to higher speeds to isolate smaller organelles, such as mitochondria and ribosomes. This sequential process allows for the stepwise separation of cellular components.

3. What are the main steps involved in the differential centrifugation process?

Ans.The main steps in differential centrifugation include: 1. Cell lysis: Cells are broken open to release their contents, often using a buffer solution. 2. Initial centrifugation: The mixture is spun at low speeds to separate the larger components, forming a pellet and supernatant. 3. Collection of supernatant: The supernatant is carefully removed for further centrifugation. 4. Subsequent centrifugation: The supernatant is spun at increasing speeds to isolate smaller organelles. 5. Final collection: Each pellet is collected separately for further analysis.

4. What types of cellular components can be isolated using differential centrifugation?

Ans.Differential centrifugation can isolate various cellular components, including nuclei, mitochondria, lysosomes, peroxisomes, and ribosomes. Each component has distinct physical properties, such as size and density, allowing them to be separated effectively through sequential centrifugation steps. This isolation is essential for studying the specific functions and interactions of these organelles.

5. What precautions should be taken during the cell fractionation process to ensure accuracy and integrity?

Ans.To ensure accuracy and integrity during cell fractionation, researchers should: 1. Use appropriate buffers to maintain pH and osmotic balance, preventing organelle damage. 2. Maintain low temperatures throughout the process to minimize enzymatic activity that could degrade cellular components. 3. Handle samples gently to avoid disrupting organelle structure. 4. Calibrate centrifuges regularly to ensure consistent speeds and forces. 5. Use proper controls and replicate experiments to validate results and ensure reproducibility.

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Sep 28, 2025 Last updated

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