To generate birefringence in artificial materials has attracted increasing attention in terms of their potential for applications in sensor, tissue engineering and 

optical devices. Silk materials with patterned structures presented unique optical features, however, effectively fabricating of structural anisotropy in silk materials to directly tailor their birefringence is still challenging. Silk fibroin birefringent fibers (SBFs) with tunable birefringence were obtained in this study via 

a strategy that combined injection technique and binary solvent-exchange-induced self assembly (BSEISA). The structural deformation of these SBFs that introduced by external stimulus such as tensile and solvent swelling was critical to their birefringence. As a result, pink, yellow, green, cyan, and purple were 

successfully achieved in the interference color of the SBFs with an exchanging solvent of 25, 55, 75, 90 wt% ethanol aqueous solution, and methanol respectively. Moreover, we respectively exchanged these SBFs against with Congo red (SBF-CR), methyl orange (SBF-MO), methylene blue (SBF-MB) and rhodamine B (SBF-RhB) solutions to produce fibers with diversity in their birefringent performance. Two types of patterns were designed and thereafter constructed by (1) SBF\SBF-CR\SBF-RhB, and (2) SBF\SBF-MB\SBF-CR. Interestingly, the patterns both displayed a letter of “A” in natural light, while displayed different letters of (1) “H” and (2) “U” in polarized light. This study demonstrated that these SBFs with unique optical and birefringent performances are anticipated  to act as sensors and code labels for optical applications.