Brief introduction to the technique of flow cytometry|同科生物-华强电子网

Flow Cytometry (FCM) is a powerful technique used for the rapid, quantitative analysis and sorting of individual cells or biological particles—such as microspheres, bacteria, or small organisms—arranged in a single file within a liquid stream. Originating in the 1960s and 1970s, modern flow cytometry has evolved into a highly advanced technology that plays a critical role across various scientific fields, including cell biology, immunology, hematology, oncology, pharmacology, genetics, and clinical diagnostics. This technology enables the detection of multiple parameters simultaneously, such as cell size, DNA content, protein levels, enzyme activity, and surface markers. By using techniques like fluorescence, light scattering, and electrical resistance, flow cytometry allows researchers to classify and isolate specific cell populations with high precision. The sorting speed can reach up to 30,000 cells per second, making it an essential tool in both research and clinical settings. Flow cytometry integrates several disciplines, including fluid dynamics, laser technology, electronics, photodetection, computer science, fluorescent chemistry, and monoclonal antibody techniques. Over the decades, it has seen continuous improvements, especially in detection accuracy, sorting efficiency, and high-throughput analysis capabilities. The history of flow cytometry dates back to the 1930s, with early developments focusing on automated cell counting and measurement. In the 1960s, key milestones were achieved, such as the creation of multi-parameter flow cytometers and the first cell sorters. The introduction of lasers in the late 1960s marked a significant advancement, enabling more detailed and accurate measurements. At its core, flow cytometry works by suspending cells in a fluid stream and passing them through a laser beam. Fluorescently labeled cells emit light when excited, which is then detected and analyzed. This data helps determine cell characteristics such as DNA content, cell cycle phase, and surface antigen expression. The scattered light also provides insights into cell size and internal complexity. For cell sorting, the system uses high-frequency oscillation to break the cell stream into droplets. These droplets are charged based on their properties, and then deflected using electric fields to separate desired cells from the rest. This process allows for the isolation of specific cell types for further study. In research, flow cytometry is widely used in cell biology to analyze the cell cycle, monitor DNA synthesis, and assess cell viability. It also plays a vital role in genetic studies, where it helps determine chromosomal DNA content and identify abnormalities. In immunology, it aids in identifying immune cell subsets and evaluating receptor expression. In oncology, it assists in diagnosing cancers by detecting abnormal DNA ploidy and monitoring treatment responses. Beyond these areas, flow cytometry has applications in hematology, microbiology, and molecular biology. As the technology continues to evolve, it is becoming more sensitive, faster, and capable of capturing detailed morphological information, further expanding its utility in scientific research and medical diagnostics.

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