Tartrazine-enabled enhanced image depth penetration with quantitative oblique back-illumination microscopy

In biological samples, optical imaging technologies are inherently limited by light scattering in thick tissues. Recently, water-soluble, strongly absorbing dye molecules, such as tartrazine, have demonstrated reversible in vivo optical transparency through refractive index matching, enhancing imaging depth in depth-resolved imaging technologies. In this study, we applied topical tartrazine–agarose solution to mouse abdominal skin, enabling optical transparency and allowing for improved imaging depth of quantitative oblique back-illumination microscopy (qOBM), a novel 3D optical technology that achieves label-free imaging based on refractive index contrast. Using an entropy cutoff metric to define imaging penetration depths, we showed a wavelength-dependent, 1.5–2× factor improvement in imaging depth compared to baseline before tartrazine was applied. Finally, we showed no residual effect on imaging depth after removal of the dye when compared to baseline abdominal skin imaging. These findings build upon previous works that demonstrate imaging depth improvements by utilizing highly absorbing, water-soluble dyes such as tartrazine, adding momentum to translating these technologies into clinical spaces in dermatology, gynecology, and surgical oncology.