In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy

Remco T. A. Megens, Sietze Reitsma, Lenneke Prinzen, Mirjam G. A. Oude Egbrink, Wim Engels, Peter J. A. Leenders, Ellen J. L. Brunenberg, Koen D. Reesink, Ben J. A. Janssen, Bart M. ter Haar Romeny, Dick W. Slaaf, Marc A. M. J. van Zandvoort*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

35 Citations (Web of Science)


In vivo (molecular) imaging of the vessel wall of large arteries at subcellular resolution is crucial for unraveling vascular pathophysiology. We previously showed the applicability of two-photon laser scanning microscopy (TPLSM) in mounted arteries ex vivo. However, in vivo TPLSM has thus far suffered from in-frame and between-frame motion artifacts due to arterial movement with cardiac and respiratory activity. Now, motion artifacts are suppressed by accelerated image acquisition triggered on cardiac and respiratory activity. In vivo TPLSM is performed on rat renal and mouse carotid arteries, both surgically exposed and labeled fluorescently (cell nuclei, elastin, and collagen). The use of short acquisition times consistently limit in-frame motion artifacts. Additionally, triggered imaging reduces between-frame artifacts. Indeed, structures in the vessel wall (cell nuclei, elastic laminae) can be imaged at subcellular resolution. In mechanically damaged carotid arteries, even the subendothelial collagen sheet (similar to 1 mu m) is visualized using collagen-targeted quantum dots. We demonstrate stable in vivo imaging of large arteries at subcellular resolution using TPLSM triggered on cardiac and respiratory cycles. This creates great opportunities for studying (diseased) arteries in vivo or immediate validation of in vivo molecular imaging techniques such as magnetic resonance imaging (MRI), ultrasound, and positron emission tomography (PET).
Original languageEnglish
Pages (from-to)10
JournalJournal of Biomedical Optics
Issue number1
Publication statusPublished - 2010


  • in vivo imaging
  • two-photon microscopy
  • subendothelial collagen
  • triggering
  • quantum dots
  • heart rate
  • respiration rate

Cite this