Two-photon excited autofluorescence imaging of human retinal pigment epithelial cells.
Degeneration of retinal pigment epithelial (RPE) cells severely impairs the visual function of retina photoreceptors. However, little is known about the events that trigger the death of RPE cells at the subcellular level. Two-photon excited autofluorescence (TPEF) imaging of RPE cells proves to be well suited to investigate both the morphological and the spectral characteristics of the human RPE cells. The dominant fluorophores of autofluorescence derive from lipofuscin (LF) granules that accumulate in the cytoplasm of the RPE cells with increasing age. Spectral TPEF imaging reveals the existence of abnormal LF granules with blue shifted autofluorescence in RPE cells of aging patients and brings new insights into the complicated composition of the LF granules. Based on a proposed two-photon laser scanning ophthalmoscope, TPEF imaging of the living retina may be valuable for diagnostic and pathological studies of age related eye diseases.
Han
Meng
M
Bindewald-Wittich
Almut
A
Holz
Frank G
FG
Giese
Guenter
G
Niemz
Markolf H
MH
Snyder
Sarah
S
Sun
Hui
H
Yu
Jiayi
J
Agopov
Michael
M
La Schiazza
Olivier
O
Bille
Josef F
JF
D000328:Adult; D000369:Aged, 80 and over; D002102:Cadaver; D006801:Humans; D066298:In Vitro Techniques; D008062:Lipofuscin / Q000378:metabolism*; D049408:Luminescent Agents / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D008875:Middle Aged; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism*
D008062:Lipofuscin; D049408:Luminescent Agents