Tanner built upon this ICP-MS with the first multiplexed assay using lanthanide metals to label DNA and cell surface markers. Inspired by Flow cytometry, in 2007 Scott D. In this type of experiment elements such as calcium within the cell could be quantified. Nomizu realized that single cells could be nebulized, dried, and ignited in plasma to generate clouds of ions which could be detected by emission spectrometry. In 1994 Tsutomu Nomizu and colleagues at Nagoya University performed the first mass spectrometry experiments of single cells. Isotope chelation, antibody tagging, cellular staining, and aerosol injection into the ICP-MS, and data analysis. įigure 1: Major steps of a CyTOF procedure. Using large numbers of probes creates new problems in analyzing the high dimensional data generated. However, this sensitivity also means trace heavy metal contamination is a concern. Mass spectrometry's sensitivity to detect different ions allows measurements of upwards of 50 targets per cell while avoiding issues with spectral overlap seen when using fluorescent probes. Ion abundances correlate with the amount of target per cell and can be used to infer cellular qualities. The remaining heavy ions from the antibody tags are quantified by Time-of-flight mass spectrometry. Abundant low weight ions generated from environmental air and biological molecules are removed using a quadrupole mass analyzer. This breaks the cells into their individual atoms and creates an ion cloud. The sample-gas mixture is focused and ignited with an argon plasma torch. Labeled cells are nebulized and mixed with heated argon gas to dry the cell containing particles. Targets are selected to answer a specific research question and are labeled with lanthanide metal tagged antibodies. CyTOF allows the quantification of multiple cellular components simultaneously using an ICP-MS detector.ĬyTOF takes advantage of immunolabeling to quantify proteins, carbohydrates or lipids in a cell. Cytometry by time of flight, or CyTOF, is an application of mass cytometry used to quantify labeled targets on the surface and interior of single cells.
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