title: Phase contrast microscopy of living cells within the whole lens: spatial correlations and morphological dynamics.
abstract: OBJECTIVE
  Images from cultured lens cells do not convey enough spatial information, and imaging of fixed lens specimens cannot reveal dynamic changes in the cells. As such, a real-time, convenient approach for monitoring label-free imaging of dynamic processes of living cells within the whole lens is urgently needed.
  
  
  METHODS
  Female Wistar rat lenses were kept in organ culture. Insulin-like growth factor-I was added to the culture medium to induce cell mitosis. A novel method of ultraviolet (UV) irradiation was used to induce cell apoptosis and fiber damage. The cellular morphological dynamics within the whole lens were monitored by inverted phase contrast microscopy. Apoptosis was assessed using a commercial kit with Hoechst 33342/YO-PRO®-1/propidium iodide (PI).
  
  
  RESULTS
  The intrinsic transparency and low-light scattering property of the rat lens permitted direct imaging of the lens epithelial cells (LECs) and the superficial fiber cells. We visualized the processes of mitosis and apoptosis of the LECs, and we obtained dynamic images of posterior fiber cells following UVA irradiation.
  
  
  CONCLUSIONS
  This method opens a new window for observing lens cells in their physiologic location, and it can be readily applied in studies on lens physiology and pathology.
authors.affiliation: Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, PR China.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Kong
authors.forename: Xiangjia
authors.initials: X
authors.lastname: Zhu
authors.forename: Shenghai
authors.initials: S
authors.lastname: Zhang
authors.forename: Jihong
authors.initials: J
authors.lastname: Wu
authors.forename: Yi
authors.initials: Y
authors.lastname: Luo
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000528:radiation effects; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology*; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation / Q000379:methods*; D008938:Mitosis / Q000187:drug effects; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D059008:Time-Lapse Imaging; D014466:Ultraviolet Rays
chemical_list: C548904:insulin-like growth factor-1, rat; D007334:Insulin-Like Growth Factor I