3-photon fluorescence imaging of sulforhodamine B-labeled elastic fibers in the mouse skin in vivo.
J Biophotonics
; 12(11): e201900185, 2019 11.
Article
en En
| MEDLINE
| ID: mdl-31276315
ABSTRACT
Elastic fibers are key constituents of the skin. The commonly adopted optical technique for visualizing elastic fibers in the animal skin in vivo is 2-photon microscopy (2 PM) of autofluorescence, which typically suffers from low signal level. Here we demonstrate a new optical methodology to image elastic fibers in animal models in vivo 3-photon microscopy (3 PM) excited at the 1700-nm window combining with preferential labeling of elastic fibers using sulforhodamine B (SRB). First, we demonstrate that intravenous injection of SRB can circumvent the skin barrier (encountered in topical application) and preferentially label elastic fibers, as verified by simultaneous 2 PM of both autofluorescence and SRB fluorescence from skin structures. Then through 3-photon excitation property characterization, we show that 3-photon fluorescence can be excited from SRB at the 1700-nm window, and 1600-nm excitation is most efficient according to our 3-photon action cross section measurement. Based on these results and using our developed 1600-nm femtosecond laser source, we finally demonstrate 3 PM of SRB-labeled elastic fibers through the whole dermis in the mouse skin in vivo, with only 3.7-mW optical power deposited on the skin surface. We expect our methodology will provide novel optical solution to elastic fiber research.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Rodaminas
/
Piel
/
Microscopía de Fluorescencia por Excitación Multifotónica
/
Elasticidad
Límite:
Animals
Idioma:
En
Revista:
J Biophotonics
Asunto de la revista:
BIOFISICA
Año:
2019
Tipo del documento:
Article
País de afiliación:
China