Analysis of living cells grown on different titanium surfaces by time-lapse confocal microscopy.
In this study we have combined fluorescence- and reflection-confocal laser scanning microscopy for the simultaneous visualization of living cells and surface topography beneath them. To this purpose we have designed a specific flow chamber and we have tested it with osteoblasts grown on an opaque, thick support, made of smooth or sandblasted titanium. Cells were loaded with Calcein-AM or tetramethylrhodamine methyl ester (TMRM), two probes employed as indicators of cell viability/morphology and mitochondrial membrane potential, respectively. Besides the acquisition of stacks of confocal sections, the system allowed also vertical views and faithful three-dimensional reconstruction of the samples. Confocal microscope implemented with our flow chamber proved to be a promising tool for time-lapse investigation of cell-biomaterial interactions.
Gatti
R
R
Department of Experimental Medicine, University of Parma, Via Volturno 39, 43100 Parma, Italy.
Orlandini
G
G
Uggeri
J
J
Belletti
S
S
Galli
C
C
Raspanti
M
M
Scandroglio
R
R
Guizzardi
S
S
D001672:Biocompatible Materials; D002478:Cells, Cultured; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure*; D012235:Rhodamines / Q000378:metabolism; D013499:Surface Properties; D014025:Titanium
D001672:Biocompatible Materials; D005452:Fluoresceins; D005456:Fluorescent Dyes; D012235:Rhodamines; C401833:tetramethylrhodamine methyl ester; D014025:Titanium; C007740:fluorexon