Biomedical Engineering Reference
In-Depth Information
FIgurE 15.5
(
See color insert.
) Normal and fibrotic human lung samples. (Top) SHG/2PEF images and (bot-
tom) transmitted-light images of serial histological sections (scale bar: 200 μm). (Top) Unstained section, with
SHG in green and 2PEF in red; (bottom) HPS staining, and Masson's trichrome staining. (a) Control lung show-
ing a bronchovascular axis and surrounding alveolar spaces with thin alveolar walls, as revealed by HPS staining
(a2). Masson's trichrome staining (a3) highlights fibrosis in green, which is seen mainly around the vessels and
the bronchovascular axis, and correlates to the SHG collagen distribution (a1, green and yellow). (b) Fibrotic lung
(idiopathic pulmonary fibrosis) showing marked architectural change with fibrosis and distorted alveolar spaces.
(c) Fibroblastic focus composed by myo-fibroblasts (c2), and surrounded by fibrosis (c3, Masson's trichrome).
SHG/2PEF image (c1) highlights the collagen distribution mainly in the periphery of the focus, with thinner fibers
in its center, and the central accumulation of fibroblasts. (From Pena, A.-M. et al. Three-dimensional investigation
and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy.
Microsc. Res.
Tech.
2007. 70:162-170. Copyright Wiley-VCH Verlag GmbH & Co. KGaA. Reproduced with permission.)
FIgurE 15.6
(
See color insert.
) SHG microscopy versus histological staining of kidney cortex. (a) Multiphoton
image of an unstained human renal implant biopsy. SHG (green color) reveals collagen fibers in the Bowman cap-
sule and the tubular interstitium. Endogenous 2PEF (red color) underlines tubules, glomeruli, and arterioles. PCT,
convoluted proximal tubules; DT, distal tubules; G, glomerulus; white arrows, arterioles. (b,c) Histological versus
(d,e) multiphoton images (same colors as for a) of serial renal sections of control mouse (b,d) and hypertensive
mouse infused with AngII for 28 days (c,e). Note that hypertension promotes the accumulation of nonfibrillar ECM
within the glomerular tuft stained with Masson's trichrome (c) (SHG silent in (e)) and the accumulation of fibril-
lar collagen in the Bowman capsule and the surrounding interstitium, similarly to the observation in the human
biopsy in (a). Scale bars: 100 μm (a), 20 μm (b,c,d,e). (Simplified from Strupler, M. et al. 2008.
J. Biomed. Optics
13:054041. With permission of SPIE.)
