Facs Purification
Facs Purification - Related Videos
15:29
Purification of Specific Cell Population by Fluorescence Activated Cell Sorting (FACS)
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Cited by 137 •
2010
For many scientific studies requiring a biological and chemical analysis of cell populations the cells must be in a high state of purity. Fluorescence activated cell sorting (FACS) is a superior method in which to obtain pure cell populations.
06:04
Murine Dermal Fibroblast Isolation by FACS
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Cited by 28 •
2016
Fibroblast behavior underlies a spectrum of clinical entities, but they remain poorly characterized, largely due to their inherent heterogeneity. Traditional fibroblast research relies upon in vitro manipulation, masking in vivo fibroblast behavior. We describe a FACS-based protocol for the isolation of mouse skin fibroblasts that does not require cell culture.
17:07
Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS): Isolation of Splenic B Lymphocytes
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2023
Source: Perchet Thibaut1,2,3, Meunier Sylvain1,2,3, Sophie Novault4, Rachel Golub1,2,3 1 Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, Paris, France 2 INSERM U1223, Paris, France 3 Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France 4 Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute, Paris, France The overall function of the immune system is to defend the body against infectious...
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
06:30
Gel Purification
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2023
Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, washing and eluting from a silica column. DNA is believed to bind to silica in the presence of high salt via a salt bridge. Following binding, DNA is washed of impurities and eluted under low salt conditions disrupting this interaction. This video goes through a step-by-step, generalized procedure for cutting out a band from the gel, gel...
FAQs
Fluorescent antibodies or dyes attach to or mark cells with characteristics relevant to the population being studied. A laser-based flow cytometer measures fluorescence from individual cells as they pass through the instrument. Those measurements allow the system to distinguish labeled populations within a heterogeneous sample and identify which cells should be collected for subsequent analysis or experiments.
Hydrodynamic focusing organizes the sample into a narrow fluid stream so cells pass through the analysis region individually. This arrangement supports cell-by-cell measurement by the laser-based flow cytometer rather than producing a combined signal from many cells. Individual analysis is essential when the goal is to recognize and isolate a particular population from a mixed sample.
After the instrument identifies cells with the desired measured characteristics, selected cells are incorporated into charged droplets. Electrostatic deflection then directs those droplets into a collection path separate from droplets containing other cells. This coupling of optical measurement with physical separation enables collection of enriched populations instead of merely recording their presence in the original sample.
The workflow begins with a heterogeneous cell sample and labeling with fluorescent antibodies or dyes. Cells are then hydrodynamically focused, examined individually by laser-based flow cytometry, and classified according to measurable characteristics. The selected cells are finally separated as charged droplets through electrostatic deflection and collected for culture, molecular analysis, functional assays, or transplantation studies.
A typical setup requires a prepared cell sample, fluorescent antibodies or dyes for labeling, a fluidic system that provides hydrodynamic focusing, and laser-based flow-cytometry equipment for individual cell analysis. It also requires droplet charging and electrostatic deflection capabilities so identified cells can be directed into separate collections. Together, these components support both recognition and recovery of target populations.
This approach is useful when investigators need to study defined or rare cell populations within complex samples. Supported applications include immune-cell characterization, rare-cell isolation, stem-cell research, and disease investigation. The enriched cells can then support molecular or functional studies, transplantation research, and the development of diagnostic or therapeutic strategies.