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Background  Activated platelets can be found on the surface of inflamed, rupture-prone and ruptured plaques as well as in intravascular thrombosis. They are key players in thrombosis and atherosclerosis. In this study we describe the construction of a radiolabeled single-chain antibody targeting the LIBS-epitope of activated platelets to selectively depict platelet activation and wall-adherent non-occlusive thrombosis in a mouse model with nuclear imaging using  in vitro  and  ex vivo  autoradiography as well as small animal SPECT-CT for  in vivo  analysis.    Methodology/Principal Findings  LIBS as well as an unspecific control single-chain antibody were labeled with  111 Indium ( 111 In) via bifunctional DTPA ( =  111 In-LIBS/ 111 In-control). Autoradiography after incubation with  111 In-LIBS on activated platelets  in vitro  (mean 3866±28 DLU/mm 2 , 4010±630 DLU/mm 2  and 4520±293 DLU/mm 2 ) produced a significantly higher ligand uptake compared to  111 In-control (2101±76 DLU/mm 2 , 1181±96 DLU/mm 2  and 1866±246 DLU/mm 2 ) indicating a specific binding to activated platelets;  P <0.05. Applying these findings to an  ex vivo  mouse model of carotid artery thrombosis revealed a significant increase in ligand uptake after injection of  111 In-LIBS in the presence of small thrombi compared to the non-injured side, as confirmed by histology (49630±10650 DLU/mm 2  vs. 17390±7470 DLU/mm 2 ;  P <0.05). These findings could also be reproduced  in vivo . SPECT-CT analysis of the injured carotid artery with  111 In-LIBS resulted in a significant increase of the target-to-background ratio compared to  111 In-control (1.99±0.36 vs. 1.1±0.24;  P <0.01).    Conclusions/Significance  Nuclear imaging with  111 In-LIBS allows the detection of platelet activation  in vitro  and  ex vivo  with high sensitivity. Using SPECT-CT, wall-adherent activated platelets in carotid arteries could be depicted  in vivo . These results encourage further studies elucidating the role of activated platelets in plaque pathology and atherosclerosis and might be of interest for further developments towards clinical application.

Activated Platelets in Carotid Artery Thrombosis in Mice Can Be Selectively Targeted with a Radiolabeled Single-Chain Antibody