Fluorescence spectroscopy and birefringence of molecular changes in maturing rat tail tendon.
Tissue remodeling during maturation, wound healing, and response to vascular stress involves molecular changes of collagen and elastin in the extracellular matrix (ECM). Two optical techniques are effective for investigating these changes--laser-induced fluorescence (LIF) spectroscopy and polarizing microscopy. LIF spectroscopy integrates the signal from both elastin and collagen cross-linked structure, whereas birefringence is a measure of only collagen. Our purpose is (1) to evaluate the rat tail tendon (RTT) spectroscopy against data from purified extracted protein standards and (2) to correlate the two optical techniques in the study of RTT and skin. Spectra from tissue samples from 27 male rats and from extracted elastin and collagen were obtained using LIF spectroscopy (357 nm). Birefringence was measured on 5-mum histological sections of the same tissue. Morphometric analysis reveals that elastin represents approximately 10% of tendon volume and contributes to RTT fluorescence. RTT maximum fluorescence emission intensity (FEI(max)), which includes collagen and elastin, increases with animal weight (R(2)=0.64). Birefringence, when plotted against weight, increases to a plateau (nonlinear correlation: R(2)=0.90), tendon having greater birefringence than skin. LIF spectroscopy and collagen fiber birefringence are shown to provide complementary measurements of molecular structure (tendon birefringence versus FEI(max) at R(2)=0.60).
Korol
Renee M
RM
University of Western Ontario, Department of Medical Biophysics, London, Ontario N6A 5C1, Canada. rkorol@uwo.ca
Finlay
Helen M
HM
Josseau
Melanie J
MJ
Lucas
Alexandra R
AR
Canham
Peter B
PB
D000375:Aging / Q000378:metabolism*; D000818:Animals; D001718:Birefringence; D003094:Collagen / Q000378:metabolism*; D004549:Elastin / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012031:Refractometry / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013623:Tail / Q000502:physiology*; D013710:Tendons / Q000502:physiology*; D014018:Tissue Distribution
D003094:Collagen; D004549:Elastin