Linear birefringence of the retinal nerve fiber layer measured in vitro with a multispectral imaging micropolarimeter

Xiang Run Huang, Robert W. Knighton

Research output: Contribution to journalArticlepeer-review

49 Scopus citations


Scanning laser polarimetry (SLP) assesses the retina nerve fiber layer (RNFL) for glaucoma diagnosis by detecting the birefringence of the peripapillary RNFL. A detailed understanding of SLP requires an accurate value for RNFL birefringence in order to relate measured retardance to RNFL thickness, but current knowledge of this value is limited. A multispectral imaging micropolarimeter of PSC'A type was used to measure the retardance in transmission of the RNFL of isolated rat retina before (living) and after (fixed) 20 min of glutaraldehyde fixation. The thickness of the nerve fiber bundles measured was then determined histologically. As previously known from reflectance measurements, in transmission the RNFL behaved as a linear retarder. The retardance of the RNFL was constant at wavelengths from 440 to 830 nm and persisted after tissue fixation. In 37 nerve fiber bundles of 8 retinas, the average RNFL birefringence was 0.23 nm/μm before and 0.19 nm/μm after fixation, with an uncertainty of 0.01 nm/μm. The wavelength independence is consistent with a mechanism of form birefringence from thin cylindrical organelles. These results allow extrapolation of previous visible wavelength measurements to the near-infrared wavelengths used by SLP and validate the use of fixed tissue for RNFL research.

Original languageEnglish (US)
Pages (from-to)199-204
Number of pages6
JournalJournal of Biomedical Optics
Issue number2
StatePublished - Apr 2002


  • Birefringence
  • Polarimetry
  • Retinal nerve fiber layer

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Radiology Nuclear Medicine and imaging
  • Atomic and Molecular Physics, and Optics
  • Radiological and Ultrasound Technology


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