Fluorescent labeling with 5-DTAF reduces collagen fiber uncrimping in loaded tendons

A fluorescent dye commonly used to image tissues under load (5-DTAF) has previously been shown to stiffen tendons. This study hypothesized that 5-DTAF staining stiffens tendons through reduced fiber sliding, altering the rate at which crimped collagen fibers straighten under load. This was tested by using reflected cross-polarized light microscopy to measure fiber crimp period of cervine extensor digitorum longus tendon specimens under axial load. Specimens were treated with either phosphate buffered saline (negative control), genipin (positive control), or 5-DTAF. In saline treated specimens, crimp period (relative to unstretched) increased at approximately 2.5 times the applied axial strain, indicating substantial fiber sliding. In both 5-DTAF and genipin treated specimens, this ratio was reduced to 1:1, indicating no fiber sliding. These results add further evidence that care should be taken when using 5-DTAF to stain tissue for studying microscale deformations in tissues.