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. CyTOF allows the quantification of multiple cellular components simultaneously using an ICP-MS detector.
CyTOF takes advantage of immunolabeling to quantify proteins, carbohydrates or lipids in a cell. Targets are selected to answer a specific research question and are labeled with lanthanide metal tagged antibodies. Labeled cells are nebulized and mixed with heated argon gas to dry the cell containing particles. The sample-gas mixture is focused and ignited with an argon plasma torch. This breaks the cells into their individual atoms and creates an ion cloud. Abundant low weight ions generated from environmental air and biological molecules are removed using a quadrupole mass analyzer. The remaining heavy ions from the antibody tags are quantified by Time-of-flight mass spectrometry.[1] Ion abundances correlate with the amount of target per cell and can be used to infer cellular qualities.[2]
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.[3][4] However, this sensitivity also means trace heavy metal contamination is a concern.[5] Using large numbers of probes creates new problems in analyzing the high dimensional data generated.[6]
Figure 1: Major steps of a CyTOF procedure. Isotope chelation, antibody tagging, cellular staining, and aerosol injection into the ICP-MS, and data analysis.
^Dmitry Bandura; Vladimir Baranov; Olga Ornatsky; Scott D. Tanner; Antonov A; Kinach R; Lou X; Pavlov S; Vorobiev S; Dick JE (August 2009). "Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry". Analytical Chemistry. 81 (16): 6813–22. doi:10.1021/ac901049w. PMID 19601617.
^Wang W, Su B, Pang L, Qiao L, Feng Y, Ouyang Y, et al. (June 2020). "High-dimensional immune profiling by mass cytometry revealed immunosuppression and dysfunction of immunity in COVID-19 patients". Cellular & Molecular Immunology. 17 (6): 650–652. doi:10.1038/s41423-020-0447-2. PMC 7186533. PMID 32346099.
^Spitzer MH, Nolan GP (May 2016). "Mass Cytometry: Single Cells, Many Features". Cell. 165 (4): 780–91. doi:10.1016/j.cell.2016.04.019. PMC 4860251. PMID 27153492.
^Gadalla R, Noamani B, MacLeod BL, Dickson RJ, Guo M, Xu W, et al. (2019). "Validation of CyTOF Against Flow Cytometry for Immunological Studies and Monitoring of Human Cancer Clinical Trials". Frontiers in Oncology. 9: 415. doi:10.3389/fonc.2019.00415. PMC 6534060. PMID 31165047.
^Olsen LR, Leipold MD, Pedersen CB, Maecker HT (February 2019). "The anatomy of single cell mass cytometry data". Cytometry. Part A. 95 (2): 156–172. doi:10.1002/cyto.a.23621. PMID 30277658.
^Palit S, Heuser C, de Almeida GP, Theis FJ, Zielinski CE (2019). "Meeting the Challenges of High-Dimensional Single-Cell Data Analysis in Immunology". Frontiers in Immunology. 10: 1515. doi:10.3389/fimmu.2019.01515. PMC 6634245. PMID 31354705.
flight, or CyTOF, is an application of mass cytometry used to quantify labeled targets on the surface and interior of single cells. CyTOF allows the quantification...
infusion and changed its name to Standard BioTools. The CyTOF, CyTOF2, Helios (CyTOF3) and CyTOF XT(4th generation) have been commercialized up to now....
Fluidigm) developing instrumental concepts and algorithmics that advance the CyTOF® line of products. He also and played a fundamental role in the development...
the reward system to impact tumor growth. Rolls has pioneered the use of CyTOF mass cytometry in the characterization of immune cell populations in the...
Canadas Top 100. 2021. p. 37. "HUPO Awards - 2019 RECIPIENTS". HUPO. 2019. "CyTOF Inventors Receive Prestigious Science and Technology Award from the Human...
Standard BioTools Press Release. 9 January 2014. "Dvs Sciences Introduces The Cytof® 2". Standard BioTools Press Release. 4 May 2013. "Mycometrix changes name...
lymphoma, colorectal cancer, and multiple myeloma. A biomarker study using CyTOF is also underway. The success of Urelumab in promoting anti-tumor immunity...
D. Tanner, Vladimir Baranov and Dmitry Bandura, she helped develop the CyTOF™ Mass Cytometer, for highly multi-parametric single cell analysis at the...
Force SMG RAF St Mawgan Search and Rescue United Kingdom Royal Air Force TOF RAF Topcliffe Flying Training Unit TOPCLIFFE United Kingdom Royal Air Force...
Archived from the original on 27 September 2016. Retrieved 26 September 2016. Tof I (1994). "Recombinant DNA technology in the synthesis of human insulin"...
alkaline pH on the β-lactoglobulin trypsin hydrolysis pattern using MALDI-TOF-MS/MS". Journal of Agricultural and Food Chemistry. 59 (5): 1572–81. doi:10...
Biotechnology. 11 (5): 450–54. doi:10.1016/S0958-1669(00)00131-2. PMID 11024362. Tof I (1994). "Recombinant DNA Technology in the Synthesis of Human Insulin"...
chips—prefabricated chips that test many markers at once,: ch 24 or MALDI-TOF The same principle applies to the proteome and the genome. High-throughput...
Take off (TOF): Initiation of takeoff power, pulling back on controls, through to 10 m (35 ft) altitude. Initial climb (ICL): End of TOF to the first...
Dokken C (October 2005). "Detection of ApoE E2, E3 and E4 alleles using MALDI-TOF mass spectrometry and the homogeneous mass-extend technology". Nucleic Acids...
use of two-dimensional GC at the separation stage, as part of a GC x GC-TOF-MS system. In contrast to environmental sample analyses, synthetic studies...
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