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21 February 2006 Use of formalin-fixed tissues for ex vivo imaging with optical coherence tomography (OCT)
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Structural and compositional analysis of normal and pathological tissues by OCT often is performed ex vivo and subsequently compared to the histology. Many of the tissues of interest require immediate fixation to prevent degradation of the sample. Frequently, samples are obtained up to a week prior to procuring images by OCT. We investigated whether fixation affects OCT image analysis by acquiring images of freshly isolated bovine ligament samples and repeating OCT imaging of the same area after fixation at 24 hours and at one week. Samples were divided into two groups: group one was fixed in 10% neutral buffered formalin for 24 hours and placed in normal saline while group two remained in formalin for one week. Tissue samples were processed for paraffin embedment and stained with Masson's trichrome or with picrosirius red. The banding pattern contrast ratio of the OCT images before and after fixation for both groups was measured and compared for possible differences. Histology was evaluated for tissue integrity and compared to the OCT images. The mean contrast ratio at time 0 was 5.41 ± 1.1 and 5.31 ± 0.6 for groups 1 and 2, respectively. Results at 1 week were slightly lower with 5.11 ± 0.3 and 5.20 ± 0.7, respectively. Statistical analysis of the data by ANOVA showed no difference in the contrast ratios with time or with treatment. This data indicates that 24 hours in formalin is sufficient to fix these small ligament samples with little effect on imaging up to one week after fixation.
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Sonya Shortkroff, Alicia Goodwin, Susanne Giattina, Bin Liu, and Mark E. Brezinski "Use of formalin-fixed tissues for ex vivo imaging with optical coherence tomography (OCT)", Proc. SPIE 6088, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IV, 60880D (21 February 2006);

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