Abstract
Activation of T lymphocytes by antigen-presenting cells (APC) results in the formation of an immunological synapse. Following contact with the target cell, key signaling and adhesion molecules polarize within minutes to hours to the T cell-APC interface. Multispectral imaging flow cytometry, a new technology which combines flow cytometry with imaging, was used to visualize and quantify the recruitment of the CD3ε and Lck signaling molecules during the evolution of an immune synapse. Using this technology, thousands of T cell/macrophage conjugates could be analyzed for each experimental time point. Following Ca++ triggered T cell activation, the dynamics of Lck and CD3ε recruitment to the synapse, analyzed by two independent methods, were comparable. However, CD3ε exhibited longer residence times (> 8 min) at the synapse than Lck.
| Original language | English |
|---|---|
| Pages (from-to) | 79-86 |
| Number of pages | 8 |
| Journal | Journal of Immunological Methods |
| Volume | 347 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - Aug 15 2009 |
| Externally published | Yes |
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Keywords
- Flow cytometry
- Fluorescence microscopy
- Imaging flow cytometry
- Immunological synapse
ASJC Scopus subject areas
- Immunology
- Immunology and Allergy
Cite this
Numbers matter : Quantitative and dynamic analysis of the formation of an immunological synapse using imaging flow cytometry. / Ahmed, Fariyal; Friend, Sherree; George, Thaddeus C.; Barteneva, Natasha; Lieberman, Judy.
In: Journal of Immunological Methods, Vol. 347, No. 1-2, 15.08.2009, p. 79-86.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Numbers matter
T2 - Quantitative and dynamic analysis of the formation of an immunological synapse using imaging flow cytometry
AU - Ahmed, Fariyal
AU - Friend, Sherree
AU - George, Thaddeus C.
AU - Barteneva, Natasha
AU - Lieberman, Judy
PY - 2009/8/15
Y1 - 2009/8/15
N2 - Activation of T lymphocytes by antigen-presenting cells (APC) results in the formation of an immunological synapse. Following contact with the target cell, key signaling and adhesion molecules polarize within minutes to hours to the T cell-APC interface. Multispectral imaging flow cytometry, a new technology which combines flow cytometry with imaging, was used to visualize and quantify the recruitment of the CD3ε and Lck signaling molecules during the evolution of an immune synapse. Using this technology, thousands of T cell/macrophage conjugates could be analyzed for each experimental time point. Following Ca++ triggered T cell activation, the dynamics of Lck and CD3ε recruitment to the synapse, analyzed by two independent methods, were comparable. However, CD3ε exhibited longer residence times (> 8 min) at the synapse than Lck.
AB - Activation of T lymphocytes by antigen-presenting cells (APC) results in the formation of an immunological synapse. Following contact with the target cell, key signaling and adhesion molecules polarize within minutes to hours to the T cell-APC interface. Multispectral imaging flow cytometry, a new technology which combines flow cytometry with imaging, was used to visualize and quantify the recruitment of the CD3ε and Lck signaling molecules during the evolution of an immune synapse. Using this technology, thousands of T cell/macrophage conjugates could be analyzed for each experimental time point. Following Ca++ triggered T cell activation, the dynamics of Lck and CD3ε recruitment to the synapse, analyzed by two independent methods, were comparable. However, CD3ε exhibited longer residence times (> 8 min) at the synapse than Lck.
KW - Flow cytometry
KW - Fluorescence microscopy
KW - Imaging flow cytometry
KW - Immunological synapse
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UR - http://www.scopus.com/inward/citedby.url?scp=67650495486&partnerID=8YFLogxK
U2 - 10.1016/j.jim.2009.05.014
DO - 10.1016/j.jim.2009.05.014
M3 - Article
VL - 347
SP - 79
EP - 86
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -