Validation of Multi-photon Auto-fluorescence Device for the Diagnosis_v2.pptx

Validation of Multi-photon Auto-fluorescence Device for the Diagnosis

1. Validation of Multi-photon Auto-fluorescence Device for the Diagnosis of Retinal Diseases and Live Functional Imaging in Vitreo-Retinal Surgery Dr. Su Xin Yi Dr. Sun Chen-Hsin Prof Lingam/ Prof Chee

2. Multi-photon Auto-fluorescence

3. Objective Development of non-linear optical fibre -optic endoscope for live in vivo retinal imaging for use in vitreo-retinal surgery: Use of Second Harmonic Generation (SHG) channel to detect pre-retinal and intra-retinal gliosis in our Proliferative Diabetic Retinopathy (PDR) murine and rat retinal detachment models. 2. Development of Two-Photon Excited Autofluorescence (TPAF) imaging toward novel retinal diagnostic in Age-related Macular Degeneration (AMD).

4. Pathogenesis of AMD Nature Reviews Drug Discovery 11, 541-559 (July 2012)

5. Lipofusin granules that accumulate in the retinal pigment epithelium from phagocytosis of shed photoreceptor outer segments Fundus Autofluorescence Imaging in Clinical Use http://www.reviewofophthalmology.com/content/d/imaging_and_diagnostic_instruments/c/22655/

6. Lipofusin Mostly consist of A2E, isoA2E, and all- trans -retinal dimer-phosphatidylethanolamine ( atRALdi -PE ) Ng KP, Gugiu B, Renganathan K, Davies MW, Gu X, et al. (2008) Retinal pigment epithelium lipofuscin proteomics. Mol Cell Proteomics 7: 1397–1405. Thematic Review Series: Lipids and Lipid Metabolism in the Eye Phospholipid meets all- trans -retinal: the making of RPE bisretinoids Janet R. Sparrow, 1 Yalin Wu , Chul Y. Kim , and Jilin Zhou

7. Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy Biomedical Optics Express Vol. 2, Issue 1, pp. 139-148 (2011) Invest Ophthalmol Vis Sci. 2014 Jul; 55(7): 4438–4447.

8. Two-photon microscopy reveals early rod photoreceptor cell damage in light-exposed mutant mice PNAS April 8, 2014 vol. 111 no. 14

9. Current Setup

10. Two photo fluorescence imaging

11. Two photo fluorescence imaging

12. Two photo fluorescence imaging

13. Challenges and next steps Optimization of TPAF imaging for retina tissue using ex-vivo sample Difficulty in focusing and finding the signal, long image acquisition time To built a combine OCT system to provide navigational scan Need for fresh animal sample Get from SERI animal facility Need substantial modification to current system to enable thru-the-pupil imaging Adaptive optics Shorter fematosecond laser source Dispersion decompensation

14. Micro-OCT images of pig retina En-face reconstruction of choroid layer Fresh pig eye Formalin fixed flat mount retina Formalin fixed flat mount Choroid