Herovici collagen stain kit

Mature collagen fibers and newly synthesized collagen fibers were identified in skin tissues using a Herovici Collagen Stain Kit (Scytek Laboratories) [56 (link),57 (link)]. Deparaffinized slides were incubated in Weigert’s iron hematoxylin for 8 min to stain nuclei, followed by rinsing with tap water and distilled water. After rinsing, slides were treated with Herovici solution for 2 min, washed, dehydrated using graded alcohols (70–100%), cleared in xylene, and mounted with DPX mounting solution (Sigma-Aldrich). Microscopic observation and imaging were conducted using a slide scanner (Motic). Newly synthesized collagen fibers were stained blue, and mature collagen fibers were stained red. The staining densities were quantitatively analyzed using ImageJ software (NIH) [58 (link)].

Mature collagen, newly generated collagen, and reticulin were identified in skin tissues using a Herovici Collagen Stain Kit (Scytek Laboratories, West Logan, UT, USA). Deparaffinized slides were incubated in Weigert’s iron hematoxylin for 8 min to stain the nuclei and then washed with tap water and DW. The slides were then treated with Herovici solution for 2 min and mounted with DPX mounting solution (Sigma-Aldrich, St. Louis, MO, USA). Slides were observed under a microscope and imaged using a slide scanner (Motic, Vancouver, Canada). Newly synthesized collagen was stained blue, whereas mature collagen was stained red. The staining densities were analyzed using ImageJ software (NIH, Maryland, MD, USA).

In skin tissue samples, we differentiated between mature collagen fibers and newly synthesized collagen fibers using a Herovici Collagen Stain Kit (Scytek Laboratories). Deparaffinized slides were incubated in Weigert’s iron hematoxylin for 8 min to highlight the nuclei, followed by washing with tap water and DW. Slides were then exposed to the Herovici solution for 2 min and sealed with DPX mounting solution. For microscopic observation and imaging, a slide scanner (Motic) was utilized. Newly synthesized collagen fibers stained blue, while mature collagen fibers appeared red [41 (link),42 (link)]. Quantitative staining analysis was conducted using ImageJ software.