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Fluorescence Activated Cell Sorting Video Lecture | Crash Course for CSIR NET Life Sciences

FAQs on Fluorescence Activated Cell Sorting Video Lecture - Crash Course for CSIR NET Life Sciences

1. What is FACS, and how does it differ from traditional cell sorting techniques?
Ans. FACS, or Fluorescence-activated cell sorting, is a specialized type of flow cytometry that allows for the sorting of cells based on their fluorescence characteristics. Unlike traditional cell sorting methods, which may rely on physical properties like size or density, FACS utilizes fluorescent markers attached to specific cell types. This method enables researchers to isolate and analyze specific populations of cells with high precision, making it particularly valuable in applications such as immunology and cancer research.
2. What are the key components of a FACS machine?
Ans. A FACS machine is composed of several key components, including a fluidics system, a laser system, detectors, and a computer system for data analysis. The fluidics system transports cells in a stream to the laser, where they are illuminated. The laser excites the fluorescent markers on the cells, and the emitted light is detected by the detectors. Finally, the computer system processes the data, allowing researchers to visualize and sort the cells based on their fluorescence intensity and characteristics.
3. What types of applications can FACS be used for in research and clinical settings?
Ans. FACS has a wide range of applications in both research and clinical settings. In research, it is commonly used for cell biology studies, immunology, and cancer research, allowing scientists to analyze immune cell populations or tumor cells. Clinically, FACS is utilized for diagnostic purposes, such as identifying specific cell types in blood samples, monitoring immune responses in patients, and assessing the effectiveness of therapies, especially in hematological malignancies.
4. What are some common fluorescent dyes used in FACS, and how do they work?
Ans. Common fluorescent dyes used in FACS include fluorescein isothiocyanate (FITC), phycoerythrin (PE), and allophycocyanin (APC). These dyes work by binding to specific cellular components, such as proteins or nucleic acids. When exposed to a specific wavelength of light from the laser, these dyes emit light at a longer wavelength, which is detected by the FACS machine. The emitted fluorescence intensity correlates with the amount of the target molecule present in the cell, enabling precise analysis.
5. What are some limitations of FACS technology?
Ans. Despite its advantages, FACS technology has some limitations. One major limitation is that it requires the cells to be in a single-cell suspension, which can be challenging for certain tissue types. Additionally, the complexity of the equipment and the need for specific fluorescent markers can limit its accessibility and increase costs. Furthermore, there is a potential for photobleaching of the fluorescent dyes, which can affect the accuracy of the results if not managed properly.
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