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Natural and synthetic hydrogel scaffolds exhibit distinct viscoelastic properties at various length scales and deformation rates. Laser Speckle Rheology (LSR) offers a novel, non-contact optical approach for evaluating the frequency-dependent viscoelastic properties of hydrogels. In LSR, a coherent laser beam illuminates the specimen and a high-speed camera acquires the time-varying speckle images. Cross-correlation analysis of frames returns the speckle intensity autocorrelation function,  g   2  ( t ), from which the frequency-dependent viscoelastic modulus,  G* ( ω ), is deduced. Here, we establish the capability of LSR for evaluating the viscoelastic properties of hydrogels over a large range of moduli, using conventional mechanical rheometry and atomic force microscopy (AFM)-based indentation as reference-standards. Results demonstrate a strong correlation between  |G* ( ω ) |  values measured by LSR and mechanical rheometry ( r  =  0.95, p  <  10   −9  ), and z-test analysis reports that moduli values measured by the two methods are identical (p > 0.08) over a large range (47 Pa – 36 kPa). In addition,  |G* ( ω ) |  values measured by LSR correlate well with indentation moduli,  E , reported by AFM ( r  =  0.92, p  <  10   −7  ). Further, spatially-resolved moduli measurements in micro-patterned substrates demonstrate that LSR combines the strengths of conventional rheology and micro-indentation in assessing hydrogel viscoelastic properties at multiple frequencies and small length-scales.

Laser Speckle Rheology for evaluating the viscoelastic properties of hydrogel scaffolds