Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM.
J Vis Exp
; (124)2017 06 24.
Article
em En
| MEDLINE
| ID: mdl-28671659
ABSTRACT
Imaging heterogeneous cellular structures using single molecule localization microscopy has been hindered by inadequate localization precision and multiplexing ability. Using fluorescent nano-diamond fiducial markers, we describe the drift correction and alignment procedures required to obtain high precision in single molecule localization microscopy. In addition, a new multiplexing strategy, madSTORM, is described in which multiple molecules are targeted in the same cell using sequential binding and elution of fluorescent antibodies. madSTORM is demonstrated on an activated T cell to visualize the locations of different components within a membrane-bound, multi-protein structure called the T cell receptor microcluster. In addition, application of madSTORM as a general tool for visualization of multi-protein structures is discussed.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Linfócitos T
/
Imunofluorescência
/
Imagem Individual de Molécula
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Microscopia de Fluorescência
Limite:
Humans
Idioma:
En
Revista:
J Vis Exp
Ano de publicação:
2017
Tipo de documento:
Article