DEPARTMENT OF BIOTECHNOLOGY;NATIONAL BRAIN RESEARCH CENTRE;DHINGRA, NARENDER, K.;VIDHYASANKAR, K.
发明人:
DHINGRA, NARENDER, K.,VIDHYASANKAR, K.
申请号：
WO2008IN00321
公开号：
WO2009047778(A1)
申请日:
2008.05.21
申请国别(地区):
世界知识产权组织国际局
年份:
2009
代理人:
摘要：
A method for specifically labeling outer and inner segments and pedicles of mammalian cone photoreceptors in vivo comprising Intravitreal injection of 2 ??l for adult anesthestized mouse (C57BL/6 J), 10 ??l for guinea pig (Duncan Hartley) of 20 ??l for monkey (Macaca mulatta) of PNA conjugated with a fluorescent probe (Rhodamine or Fluorescein) at a concentration of 0.005%, 0.01%, 0.02%, 0.05% or 0.5% (only 0.5% for guinea pig and monkey) with a glass electrode with tip diameter of ~ 1 ??m or an insulin syringe posterior to the limbus into the vitreous of the eye, Removing the eye under anesthesia at various time points (mouse: 0.5, 1, 2, 4 and 8 hours; 1, 2, 4, 7 and 10 days; 1 and 3 months; guinea pig and monkey: 30 minutes) after the PNA injection, Hemisecting and fixing the posterior eye-cup in 4% paraformaldehyde for 1 hour at 4°C, Flat mounting the retina on a glass slide or cryosectioning, and observing PNA labeling with a fluorescence microscope.
