Title :
Quantification of Cy-5 siRNA signal in the intra-vital multi-photon microscopy images
Author :
Chen, Aaron ; Dogdas, B. ; Mehta, Sharad ; Haskell, K. ; Ng, Bryan ; Keough, E. ; Howell, B. ; Meacham, D.A. ; Aslamkhan, A.G. ; Davide, J. ; Stanton, M. ; Bagchi, Arun ; Sepp-Lorenzino, L. ; Weikang Tao
Author_Institution :
Inf. IT, Merck Res. Labs. IT, Rahway, NJ, USA
fDate :
Aug. 28 2012-Sept. 1 2012
Abstract :
Transgenic mice with Tie2- green fluorescent protein (GFP) are used as a model to study the kinetic distribution of the Cy5-siRNA delivered by lipid nanoparticles (LNP) into the liver. After the mouse is injected with the LNP, it undergoes a procedure of intra-vital multi-photon microscopy imaging over a period of two hours, during which the process for the nanoparticle to diffuse into the hepatocytes from the vasculature system is monitored. Since the images are obtained in-vivo, the quantification of Cy5 kinetics suffers from the moving field of view (FOV). A method is proposed to register the sequence of images through template matching. Based on the semi-automatic segmentations of the vessels in the common FOV, the registered images are segmented into three regions of interest (ROI) in which the Cy5 signals are quantified. Computation of the percentage signal strength in the ROIs over time allows for the analysis of the diffusion of Cy5-siRNA into the hepatocytes, and helps demonstrate the effectiveness of the Cy5-siRNA delivery vehicle.
Keywords :
RNA; biomedical optical imaging; blood vessels; cellular biophysics; fluorescence; image registration; image segmentation; image sequences; lipid bilayers; medical image processing; molecular biophysics; nanomedicine; nanoparticles; proteins; Cy-5 siRNA signal quantification; Cy5-siRNA delivery vehicle; Cy5-siRNA diffusion; Cy5-siRNA kinetic distribution; Tie2- green fluorescent protein; hepatocyte; image registration; image sequence; intra-vital multiphoton microscopy image; intra-vital multiphoton microscopy imaging; lipid nanoparticle; liver; percentage signal strength; template matching; transgenic mice; vasculature system; vessel semi-automatic segmentation; Image segmentation; Kinetic theory; Liver; Mice; Microscopy; USA Councils; Animals; Carbocyanines; Green Fluorescent Proteins; Imaging, Three-Dimensional; Mice; Mice, Transgenic; Microscopy, Fluorescence, Multiphoton; RNA, Small Interfering; Signal Processing, Computer-Assisted;
Conference_Titel :
Engineering in Medicine and Biology Society (EMBC), 2012 Annual International Conference of the IEEE
Conference_Location :
San Diego, CA
Print_ISBN :
978-1-4244-4119-8
Electronic_ISBN :
1557-170X
DOI :
10.1109/EMBC.2012.6346773
