Three-dimensional optical coherence tomography imaging of retinal sheet implants in live rats.
OBJECTIVE
To obtain three-dimensional images from retinal transplants in live animals and evaluate the placement and structural quality of the transplants.


METHODS
Donor retinal sheets were isolated from E19 fetuses of transgenic rats expressing human alkaline phosphatase (hPAP), and transplanted to the subretinal space of 19-56 days old S334ter-3 rat recipients with fast retinal degeneration (average age at surgery 32 days). A total of 143 rats were imaged 1 day to 2.8 months after surgery, using a Fourier-domain optical coherence tomography (FDOCT) system, with an axial resolution of 3.5 microm. The CCD A-line integration time was set at 200 micros for better visualization of degenerated retina. After targeting the transplant area, 139 or 199 consecutive slices were scanned. Projection images and movies of the retinal transplant area were computed and later compared with histology.


RESULTS
OCT scans identified 137 of 141 transplants as a thickening of the degenerated retina. OCT indicated the laminar structure of the transplants and surgical defects, such as RPE/choroid damage with an accuracy rate between 83 and 99%. Three-dimensional projections showed the transplant position in the retina in relation to the optic disc. Histology of transplants by hPAP and hematoxylin-eosin staining was correlated with the OCT results.


CONCLUSIONS
Optical coherence tomography is an excellent tool to image retinal layers in a live rat. This procedure helps to evaluate the placement and quality of the transplants in the living eye.
Seiler
Magdalene J
MJ
Department of Anatomy & Neurobiology, University of California, Reeve-Irvine Research Center, Sue and Bill Gross Stem Cell Research Center, Irvine, CA, United States.
Rao
Bin
B
Aramant
Robert B
RB
Yu
Lingfeng
L
Wang
Qiang
Q
Kitayama
Eric
E
Pham
Sylvia
S
Yan
Fengrong
F
Chen
Zhongping
Z
Keirstead
Hans S
HS
D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D000818:Animals; D005583:Fourier Analysis; D058851:GPI-Linked Proteins; D006085:Graft Survival / Q000502:physiology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012160:Retina / Q000166:cytology / Q000502:physiology* / Q000637:transplantation*; D012162:Retinal Degeneration / Q000601:surgery*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000502:physiology / Q000637:transplantation; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000502:physiology / Q000637:transplantation; D012815:Signal Processing, Computer-Assisted; D013194:Staining and Labeling; D041623:Tomography, Optical Coherence / Q000379:methods*
D058851:GPI-Linked Proteins; D007527:Isoenzymes; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental