{"id":177,"date":"2020-06-28T22:54:14","date_gmt":"2020-06-29T03:54:14","guid":{"rendered":"https:\/\/www.cd-bioparticles.com\/blog\/?p=177"},"modified":"2020-06-28T22:54:14","modified_gmt":"2020-06-29T03:54:14","slug":"what-you-have-to-know-about-magnetic-activated-cell-sorting","status":"publish","type":"post","link":"https:\/\/www.cd-bioparticles.com\/blog\/applications\/what-you-have-to-know-about-magnetic-activated-cell-sorting\/","title":{"rendered":"What You Have to Know about Magnetic-activated Cell Sorting"},"content":{"rendered":"\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"576\" src=\"\/blog\/wp-content\/uploads\/2020\/06\/What-You-Have-to-Know-about-Magnetic-activated-Cell-Sorting-1-1024x576.jpg\" alt=\"\" class=\"wp-image-178\" srcset=\"\/blog\/wp-content\/uploads\/2020\/06\/What-You-Have-to-Know-about-Magnetic-activated-Cell-Sorting-1-1024x576.jpg 1024w, \/blog\/wp-content\/uploads\/2020\/06\/What-You-Have-to-Know-about-Magnetic-activated-Cell-Sorting-1-300x169.jpg 300w, \/blog\/wp-content\/uploads\/2020\/06\/What-You-Have-to-Know-about-Magnetic-activated-Cell-Sorting-1-768x432.jpg 768w, \/blog\/wp-content\/uploads\/2020\/06\/What-You-Have-to-Know-about-Magnetic-activated-Cell-Sorting-1.jpg 1920w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p>Magnetic-activated\ncell sorting (MACS) is a highly specific cell sorting technique that integrates\nimmunology, cell biology, and magnetic mechanics. Its high specificity comes\nfrom the specific recognition of antibody antigens. MACS technology has become\nthe standard method of cell sorting, from laboratory to clinic, from small\nscale to large scale, from common cells to rare cells and complex cell subsets,\nfrom human and mouse cells to other cell lines. MACS technology provides a\nmethod of high-quality cell sorting in every laboratory.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>The principle of cell separation by\nimmunomagnetic beads method<\/li><\/ul>\n\n\n\n<p>The\nseparation of cells by <a href=\"https:\/\/www.cd-bioparticles.com\/product\/affinity-magnetic-particles-list-17.html\">immunomagnetic\nbeads<\/a> method is based on the fact that the cell surface antigen can\ncombine with the specific monoclonal antibody conjugated with magnetic beads.\nIn the external magnetic field, the cells labeled with magnetic beads by\nantibodies are adsorbed and remain in the magnetic field. The cells without\nthis surface antigen are not labeled because they can not bind to the specific\nmonoclonal antibodies conjugated with magnetic beads and do not stay in the\nmagnetic field, so the cells can be separated.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Magnetic cell labeling mode<\/li><\/ul>\n\n\n\n<p>The\nmost important thing in magnetic cell sorting using MACS technology is\nhigh-quality labeling. The labeling of positive cells should be enhanced as\nmuch as possible and the background staining should be weakened. There are two\nbasic ways of magnetic labeling: direct labeling and indirect labeling.<\/p>\n\n\n\n<p>1.\nDirect magnetic cell labeling<\/p>\n\n\n\n<p>Direct\nlabeling is the fastest and most specific magnetic labeling method. At present,\nthere are a variety of directly labeled microbeads for sorting human, mouse,\nrat, and non-human primate cells.<\/p>\n\n\n\n<p>2.\nIndirect magnetic cell labeling<\/p>\n\n\n\n<p>Indirect\nmagnetic cell labeling requires a combination of monoclonal or polyclonal\nantibodies and indirect labeling microbeads. Unbound antibodies, biotinylated\nantibodies, or fluorescein-labeled antibodies can be used as the primary antibody\nfor labeling cells, and then anti-immunoglobulin beads, <a href=\"https:\/\/www.cd-bioparticles.com\/product\/biotin-list-138.html\">anti-biotin<\/a>\nor <a href=\"https:\/\/www.cd-bioparticles.com\/product\/streptavidin-list-149.html\">streptavidin\nmicrobeads<\/a> and anti-fluorescein microbeads can be used as the\nsecondary antibody to magnetically label cells.<\/p>\n\n\n\n<p>Almost\nany monoclonal or polyclonal antibody against any cells of any strains can be\nused for indirect labeling. Indirect labeling is mainly used in the following\nsituations: when there are no direct labeling beads; multiple types of cells\nneed to be sorted or removed with a mixture of several antibodies at the same\ntime; indirect labeling has an amplified effect, so it can be used in sorting\ntarget cells with weak antigen expression; when homemade antibodies or ligands\nare used.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Sorting strategy<\/li><\/ul>\n\n\n\n<p>There\nare two basic sorting strategies: positive selection strategy and depletion\nstrategy. The sequential selection strategy is the combination of the two basic\nsorting strategies or the combined use of multi-selected microbeads, so as to\nrealize the sorting of cell subsets.<\/p>\n\n\n\n<p>1.\nPositive selection strategy<\/p>\n\n\n\n<p>In the\npositive selection, the target cells were magnetically labeled and directly\nsorted as positive marker components. The sorted cells do not need to remove\nmagnetic beads and can be used for culture or follow-up operation immediately.\nThe magnetically labeled target cells can be enriched 10000 times by this\nmethod. The advantages of positive selection strategy are high purity, high\nrecovery rate, rapid and simple operation.<\/p>\n\n\n\n<p>2.\nDepletion strategy<\/p>\n\n\n\n<p>Depletion\nstrategy is a method of removing non-target cells from the cell mixture after\nmagnetic labeling, that is, unmagnetically labeled cells are target cells. The\nscope of application of the de-sorting strategy: removal of unwanted cells;\nlack of specific antibodies against target cells (such as tumor cells); no need\nfor antibodies to bind with target cells, that is, cells are not stimulated\n(e.g. functional analysis of T cells, B cells, NK cells); part of the\nsequential selection.<\/p>\n\n\n\n<p>3.\nSequential selection strategy<\/p>\n\n\n\n<p>The\ncombination of two or more basic selection strategies is mainly used to sort\ncell subsets or to obtain highly pure and rare cells.<\/p>\n\n\n\n<p>(1)\nDepletion followed by positive selection<\/p>\n\n\n\n<p>The\nsorting of cell subsets can first magnetically label the non-target cells,\nremove them, and then carry out magnetic labeling and positive selection for\nthe negative components. Scope of application: in the cell suspension,\nnon-target cells also express antigens used for positive selection of target\ncells, so it is necessary to remove this group of non-target cells; if you want\nto sort very rare cells, remove non-target cells from the cell suspension\nfirst, and carry out positive selection on the basis of enriched cells, high\npurity target cells can be obtained.<\/p>\n\n\n\n<p>(2)\nMultisort strategy<\/p>\n\n\n\n<p>Multisort\nstrategy is a technique for magnetic sorting of cells based on a variety of\nsurface markers. In multiple sorting, first of all, the target cells were\nlabeled with multi-selective microbeads, and the positive sorting of the first\nparameter was carried out. The cells are then incubated with a dissociation\nreagent, which can enzymatically dissociate the beads from the antibody. The\npositive cells were then sorted by magnetic labeling of antibody-bead complex\nagainst another cell surface marker. These cells can be sorted again for\npositive selection or depletion.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Important indexes of magnetic\nseparation of cells.<\/li><\/ul>\n\n\n\n<p>Purity\nand yield depend on the specificity of the monoclonal antibody conjugated to\nthe beads and the size of the magnetic beads. However, large beads affect cell\nactivity and cannot be detected directly by flow cytometry.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Advantages and disadvantages of\nmagnetic beads of different sizes.<\/li><\/ul>\n\n\n\n<p>1.\nSmall magnetic beads (~50 nm)<\/p>\n\n\n\n<p>Advantages:<\/p>\n\n\n\n<p>(1) It\nis mild to the cells and does not affect the subsequent culture of the isolated\ncells.<\/p>\n\n\n\n<p>(2) It\ncan be detected directly by flow cytometry without affecting the scattered\nlight.<\/p>\n\n\n\n<p>(3) The\nmagnetic beads occupy only a small part of the cell surface.<\/p>\n\n\n\n<p>Disadvantages:<\/p>\n\n\n\n<p>(1) A\nstrong magnetic field is needed to separate cells.<\/p>\n\n\n\n<p>(2) The\nseparation speed is very slow and the yield is not high.<\/p>\n\n\n\n<p>(3) A\ndisposable separation column would be used instead of an ordinary test tube.<\/p>\n\n\n\n<p>(4) The\ncost is high.<\/p>\n\n\n\n<p>2.\nLarge magnetic beads (1200-4500 nm)<\/p>\n\n\n\n<p>Advantages:<\/p>\n\n\n\n<p>(1) The\ntechnology is simple and the separation can be done in the test tube.<\/p>\n\n\n\n<p>(2) It\nis easy to increase or decrease the number of involved cells.<\/p>\n\n\n\n<p>(3) The\nseparation speed is fast and the yield is high.<\/p>\n\n\n\n<p>(4) The\ncost is low.<\/p>\n\n\n\n<p>Disadvantages:<\/p>\n\n\n\n<p>(1) It\nexerts mechanical pressure on cells and affects their biological activity,\nwhich is not conducive to isolation and culture.<\/p>\n\n\n\n<p>(2)\nCell purity is low.\n\n(3) It is easy to\nblock the nozzle of flow cytometry.\n\n\n\n<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Magnetic-activated cell sorting (MACS) is a highly specific cell sorting technique that integrates immunology, cell biology, and magnetic mechanics. Its<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[7],"tags":[45,4],"class_list":["post-177","post","type-post","status-publish","format-standard","hentry","category-applications","tag-macs","tag-magnetic-particles"],"_links":{"self":[{"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/posts\/177","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/comments?post=177"}],"version-history":[{"count":1,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/posts\/177\/revisions"}],"predecessor-version":[{"id":179,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/posts\/177\/revisions\/179"}],"wp:attachment":[{"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/media?parent=177"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/categories?post=177"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.cd-bioparticles.com\/blog\/wp-json\/wp\/v2\/tags?post=177"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}