Second harmonic generation imaging of the pig lamina cribrosa using a scanning laser ophthalmoscope-based microscope

Lasers Med Sci. 2009 Sep;24(5):787-92. doi: 10.1007/s10103-008-0641-4. Epub 2009 Feb 18.

Abstract

We describe a novel scanning laser ophthalmoscope (SLO)-based on a video-rate second harmonic generation imaging microscope. A titanium:sapphire femtosecond laser was coupled to a modified SLO. The laser beam was scanned over the sample, and the light produced by second harmonic generation (SHG) was collected for imaging at video-speed. The device was used for imaging the lamina cribrosa (LC) of enucleated pig eyes. A resolution comparable to that of commercial multiphoton microscopes was reached. The SHG images were used for determining the average pore size of the LC determined from the images; the pressure dependence of the pore size was studied by the artificial increasing of the hydrostatic pressure in the eye. A pressure increase of 44.3 mmHg enlarged the average pore size of 62 analyzed pores by a statistically significant amount. The relative pore growth was measured at four different pressure levels in 25 pores. The pressure was increased in 15 mmHg steps. A general tendency for monothonic growth was observed, although single pores grew by no means linearly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Image Processing, Computer-Assisted
  • Intraocular Pressure
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods
  • Models, Animal
  • Ophthalmoscopes
  • Ophthalmoscopy / methods*
  • Optic Disk / anatomy & histology*
  • Optic Disk / physiology
  • Sus scrofa / anatomy & histology*