Slices were embedded in Vectastain (Vector Laboratories Ltd) and viewed under a laser-scanning confocal microscope (Leica Microsystems, Wetzlar, Germany).
Sections were viewed under a laser-scanning confocal microscope (Leica DMIRE 2, TCS SL, Leica Microsystems, Heidelberg, Germany) for a 3D study of the skin vessels' structure.
After three washes for 15 min with PBS containing 0.1% Tween 20, the cells were then counterstained with TOTO-3 (Invitrogen) and viewed under a laser-scanning confocal microscope (LSM 510, Carl Zeiss, Wetzlar, Germany).
The sections were mounted and viewed under a confocal laser scanning microscopy TCS SP2 (Leica TCS SP2, Van Hopplynus, Belgium).
Sections were coverslipped in DAPI-mounting media (Vector Laboratories, Burlingame, CA, U.S.A .. Immunofluorescent-labeled sections were viewed under a confocal laser scanning microscope (Zeiss LSM610 Meta, Cambridge, United Kingdom) equipped with blue diode (405 nm), argon (458, 477, 488, and 514 nm), and helium/neon (546 and 633 nm) lasers.
Samples were viewed under a confocal laser-scanning microscope (LSM510; Zeiss, Jena, Germany).
For immunofluorescence studies, cultures were incubated with primary antibodies, followed by fluorescent secondary antibody (Alexa Fluor 488 anti-mouse IgG (1∶200, Molecular Probe, Eugene, OR) or Alexa Fluor 594 anti-rabbit IgG (1∶200) for 1 hr at RT and viewed under an Olympus laser confocal microscope.
and viewed under a Zeiss confocal laser scanning microscope LSM510 (Carl Zeiss GmbH, Jena, Germany).
