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. 2014 Oct 31;9(10):e111203.
doi: 10.1371/journal.pone.0111203. eCollection 2014.

Retinal thickness measurement obtained with spectral domain optical coherence tomography assisted optical biopsy accurately correlates with ex vivo histology

Lee R Ferguson  1 , Sandeep Grover  1 , James M Dominguez 2nd  2 , Sankarathi Balaiya  1 , Kakarla V Chalam  1
Affiliations

Retinal thickness measurement obtained with spectral domain optical coherence tomography assisted optical biopsy accurately correlates with ex vivo histology

Lee R Ferguson et al. PLoS One. .

Abstract

Background: This study determines 'correlation constants' between the gold standard histological measurement of retinal thickness and the newer spectral-domain optical coherence tomography (SD-OCT) technology in adult C57BL/6 mice.

Methods: Forty-eight eyes from adult mice underwent SD-OCT imaging and then were histologically prepared for frozen sectioning with H&E staining. Retinal thickness was measured via 10x light microscopy. SD-OCT images and histological sections were standardized to three anatomical sites relative to the optic nerve head (ONH) location. The ratios between SD-OCT to histological thickness for total retinal thickness (TRT) and six sublayers were defined as 'correlation constants'.

Results: Mean (± SE) TRT for SD-OCT and histological sections was 210.95 µm (± 1.09) and 219.58 µm (± 2.67), respectively. The mean 'correlation constant' for TRT between the SD-OCT and histological sections was 0.96. The retinal thickness for all sublayers measured by SD-OCT vs. histology were also similar, the 'correlation constant' values ranged from 0.70 to 1.17. All SD-OCT and histological measurements demonstrated highly significant (p<0.01) strong positive correlations.

Conclusion: This study establishes conversion factors for the translation of ex vivo data into in vivo information; thus enhancing the applicability of SD-OCT in translational research.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. C57BL/6 mouse retina showing the reference points for SD-OCT and histological measurements.
Dotted circle – optic nerve head (ONH); black solid circle – 1 mm diameter central area surrounding the ONH; inferior, middle and superior reference points, each 200 µm apart, both to the left and the right of the ONH.
Figure 2
Figure 2. Cross-sectional views of the C57BL/6 mouse retina by SD-OCT (left) and histology (right).
OS/IS/ELM, outer segment/inner segment/external limiting membrane; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL/GC, inner plexiform layer/ganglion cell; RNFL, retinal nerve fiber layer.
Figure 3
Figure 3. Association between mean SD-OCT and histological thickness with correlation constants >1.
Scatter plot points refer to all observations for outer nuclear layer and retinal nerve fiber layer.
Figure 4
Figure 4. Association between mean SD-OCT and histological thickness with correlation constants <1.
Scatter plot points refer to all observations for outer segment/inner segment/external limiting membrane, outer plexiform layer, inner nuclear layer, inner plexiform/ganglion cell layers, and total retinal thickness.
See this image and copyright information in PMC

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