Mapping Cell Distribution in Amorphous Scaffolds using Confocal Microscopy
Jirun Sun, Nancy Lin, Marcus T. Cicerone, Sheng Lin-Gibson
One common method for probing cell-material interactions is direct visualization via confocal laser scanning microscopy, a technique widely used for studying biological systems. However, confocal microscopy has limited depth penetration for imaging cells in a typical 3D tissue engineering scaffold, in part due to light scattering from the polymer. Through the use of optically clear scaffolds, we have demonstrated that imaging through scaffolds can be achieved down to approximately 1 mm depth. With this imaging capability, we have developed methods for quantifying the cell number and spatial distribution within a 3D scaffold of known structure. Two different imaging approaches are presented, one of which is based on taking x-y slices of the entire scaffold and counting the cells as a function of scaffold depth, and the other is based on collecting sample volumes at defined and sequential positions and counting the cells within those volumes (stack of images). The cell spatial distribution can be assessed from these data. Detailed methods for sampling, validation, and quantification will be discussed.
Proceedings of the ACS Division of Polymeric Materials: Science & Engineering, 234th National Meeting
August 19-23, 2007
Boston, MA, US
ACS Division of Polymeric Materials: Science & Engineering, 234th National Meeting
, Lin, N.
, Cicerone, M.
and Lin-Gibson, S.
Mapping Cell Distribution in Amorphous Scaffolds using Confocal Microscopy, Proceedings of the ACS Division of Polymeric Materials: Science & Engineering, 234th National Meeting, Boston, MA, US, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=854045
(Accessed February 27, 2024)