A new method for quantitative cellular imaging on 3-D scaffolds using fluorescence microscopy.
This paper presents a new image processing technique for estimating cell numbers and contours in three-dimensional (3-D) microfabricated scaffolds. The method is based on a statistical approach, and utilizes the extreme value theory, which assumes that cells in the image field are rare, high-intensity events. Confocal microscopy images of fibroblasts on 3-D structures were processed using the method, and the resulting data on cell numbers was compared with countings obtained using a Burker chamber. The results were identical to within a few percent.
Santarelli
Maria Filomena
MF
Italian National Research Council, Institute of Clinical Physiology, Pisa, Italy.
Sani
Lorenzo
L
Ahluwalia
Arti
A
Vozzi
Giovanni
G
Landini
Luigi
L
De Rossi
Danilo
D
D016475:3T3 Cells; D000465:Algorithms; D000818:Animals; D002452:Cell Count / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D048429:Cell Size; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008567:Membranes, Artificial; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D023822:Tissue Engineering / Q000379:methods
D008567:Membranes, Artificial