Abstract
Purpose :
To visualize influences of monocyte chemoattractant protein-1 (MCP-1) on the immune cells in the subconjunctival tissue using two-photon laser scanning microscope.
Methods :
LysM-eGFP mice (gene-targeted mice expressing enhanced green fluorescent protein under the control of the endogenous lysozyme M promoter) were anesthetized by isoflurane. Vessels were visualized by intravenous injection of 70 kDa of Texas red–conjugated dextran. Using two-photon laser scanning microscope, three-dimensional images of the subconjunctival tissue were acquired every 1 minute for 30 minutes through phosphate-buffered saline with or without 10µg/ml MCP-1 in custom-made eyecup. Raw imaging data were processed and analyzed with Imaris software.
Results :
Intravital green-labeled LysM-eGFP-positive cells and red-labeled vessels were successfully visualized using two-photon laser scanning microscope. Compared with the control, the number of LysM-eGFP-positive cells in the subconjunctival tissue increased in the presence of MCP-1. Moreover, the motility LysM-eGFP-positive cells was enhanced under the influence of MCP-1.
Conclusions :
Four-dimensional imaging using two-photon microscope indicated MCP-1 affected the cellular dynamics in the subconjunctival tissue. The evaluation of cellular dynamics in the subconjunctival tissue using this technique may lead to the understanding of the wound healing process after filtration surgery.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.