Effects of mechanical indentation on diffuse reflectance spectra, light transmission, and intrinsic optical properties in ex vivo porcine skin.

BACKGROUND AND OBJECTIVES: Mechanical indentation has been shown to increase light transmission through turbid tissue. In this study, we investigated the effects of localized indentation on the optical properties of ex vivo porcine skin specimens by dynamically monitoring diffuse reflectance spectra, light transmission, and applied load while controlling tissue thickness. STUDY DESIGN/METHODS: A custom-built diffuse reflectance spectroscopy (DRS) system was used to capture diffuse reflectance spectra from tissue specimens undergoing indentation. The DRS probe was designed to perform both optical sensing and tissue indentation. A mechanical load frame was used to dynamically control probe displacement and resultant specimen thickness change while recording applied load. Diffuse reflectance spectra, as well as light transmission at 630 nm, were recorded during stress relaxation tests where tissue specimens were displaced to and held at a final thickness. Tissue optical properties were extracted from reflectance spectra using a previously established look-up table (LUT) approach. RESULTS: Indentation increased light transmission through tissue during linear displacement, and continued to increase transmission during subsequent stress relaxation at constant tissue thickness. The magnitude of relative transmission increases was shown to be a function of bulk tissue compressive strain (relative thickness change). Reduced scattering coefficients calculated from the LUT at 630 nm decreased during stress relaxation, with the relative decrease in scattering also depending strongly on tissue compressive strain. Reduced scattering coefficients decreased by 12.0 ± 4.7% at 0.44 ± 0.022 compressive strain, and reduced by 35.6 ± 1.3% at 0.71 ± 0.01 compressive strain. CONCLUSION: DRS can be used to capture transient changes in intrinsic tissue optical properties during mechanical loading. Mechanical indentation modifies tissue optical properties and may be harnessed as a minimally-invasive optical clearing technique to improve optical diagnostics and therapeutics.

Mechanical tissue optical clearing technique increases imaging resolution and contrast through ex vivo porcine skin.

BACKGROUND AND OBJECTIVES: Mechanical tissue optical clearing permits light delivery deeper into turbid tissue, which may improve current optical diagnostics and laser-based therapeutic techniques. We investigated the effects of localized compression on brightfield imaging through ex vivo porcine skin by evaluating resolution and contrast of a target positioned beneath native, mechanically compressed, or chemically cleared specimens. We also evaluated the effects of indentation on dynamic tissue thickness and light transmission. STUDY DESIGN/METHODS: A 5 mm diameter, hemispherically tipped, manual load transducer was used to compress specimens using 2-44 N for 60 seconds. Chemically cleared specimens were immersed for 1 hour in glycerol or dimethyl sulfoxide. A USAF 1951 resolution target was positioned beneath specimens and imaged using brightfield microscopy. Resolution and contrast of target features were analyzed. In separate experiments, a mechanical test instrument was used to compress and hold specimens at a final thickness while measuring applied load and light transmission. RESULTS: Image intensity profiles showed that while uncompressed skin did not allow resolution of any target, localized compression allowed maximum resolution up to a line width of 173 ± 21 µm. Mechanical clearing achieved up to four times higher maximum resolution and 2-3 times higher contrast sensitivity than chemical immersion. Resolving capability was highly correlated with compressive tissue strain. Light transmission increased during tissue compression, but also increased while holding final thickness constant. CONCLUSION: Localized compression is an effective technique for increasing resolution and contrast of target features through tissue and may improve light-based diagnostics. Thickness reduction and other mechanisms appear to contribute to this effect.