Dynamic in vivo imaging of microvasculature and perfusion by miniaturized confocal laser microscopy.
BACKGROUND
Microvasculature and associated pathologies mandate dynamic imaging. We evaluated a novel miniaturized confocal laser scanning probe for in vivo visualization of blood vessels, blood flow, cell tracking and perfusion in both healthy rodents and disease models.


METHODS
The hand-held confocal microscopy system allowed a 500- to 2,400-fold magnification at a dynamically variable imaging depth. Different intravital stains were used alone or in combination for tissue, nuclear, plasma and vascular endothelial cell staining and for blood flow visualization, and targeted staining for individual cell populations.


RESULTS
Precision optical sectioning yielded high-resolution images in vivo. Leucocyte-endothelium interactions in brain microvasculature were followed in serial sections. A microthrombosis was identified after sequential injection of FITC-labelled erythrocytes, FITC-dextran and acriflavine. Glomerular alterations were visualized in the MRL/lprmouse model of lupus nephritis.


CONCLUSIONS
Intravital confocal microscopy with a miniaturized hand-held probe combines high-resolution subsurface imaging in real time for dynamic visualization of vessels, cells, blood flow and associated pathologies, permitting a truly comprehensive vascular imaging in vivo at the cellular level.
Goetz
M
M
First Medical Clinic, University of Mainz, Mainz, Germany. mgoetz@mail.uni-mainz.de
Thomas
S
S
Heimann
A
A
Delaney
P
P
Schneider
C
C
Relle
M
M
Schwarting
A
A
Galle
P R
PR
Kempski
O
O
Neurath
M F
MF
Kiesslich
R
R
D000818:Animals; D001783:Blood Flow Velocity; D001921:Brain / Q000098:blood supply; D002450:Cell Communication; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology; D005260:Female; D005456:Fluorescent Dyes; D005849:Gerbillinae; D021621:Imaging, Three-Dimensional; D007249:Inflammation / Q000473:pathology; D020767:Intracranial Thrombosis / Q000473:pathology / Q000503:physiopathology; D007962:Leukocytes / Q000166:cytology; D008181:Lupus Nephritis / Q000473:pathology / Q000503:physiopathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019463:Mice, Inbred MRL lpr; D008833:Microcirculation / Q000502:physiology*; D018613:Microscopy, Confocal / Q000379:methods*; D055806:Microvessels / Q000033:anatomy & histology* / Q000502:physiology*; D008904:Miniaturization; D013927:Thrombosis / Q000473:pathology / Q000503:physiopathology
D005456:Fluorescent Dyes