In situ quantification of pancreatic beta-cell mass in mice.
Tracing changes of specific cell populations in health and disease is an important goal of biomedical research. The process of monitoring pancreatic beta-cell proliferation and islet growth is particularly challenging. We have developed a method to capture the distribution of beta-cells in the intact pancreas of transgenic mice with fluorescence-tagged beta-cells with a macro written for ImageJ (rsb.info.nih.gov/ij/). Following pancreatic dissection and tissue clearing, the entire pancreas is captured as a virtual slice, after which the GFP-tagged beta-cells are examined. The analysis includes the quantification of total beta-cell area, islet number and size distribution with reference to specific parameters and locations for each islet and for small clusters of beta-cells. The entire distribution of islets can be plotted in three dimensions, and the information from the distribution on the size and shape of each islet allows a quantitative and qualitative comparison of changes in overall beta-cell area at a glance.
Kim
Abraham
A
Department of Medicine, University of Chicago, USA.
Kilimnik
German
G
Hara
Manami
M
D000818:Animals; D048708:Cell Growth Processes / Q000502:physiology; D004210:Dissection; D049452:Green Fluorescent Proteins / Q000032:analysis; D050417:Insulin-Secreting Cells / Q000166:cytology*; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D057054:Molecular Imaging / Q000379:methods*
D049452:Green Fluorescent Proteins