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Figure 15.5 Normal and fibrotic human lung samples. (Top) SHG/2PEF images and (bottom) 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 showing a bronchovascular axis and
surrounding alveolar spaces with thin alveolar walls, as revealed by HPS staining (a2). Masson's trichrome stain-
ing (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 accumu-
lation 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 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 capsule 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) multi-
photon 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 fibrillar 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.)
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