นวัตกรรมใหม่

Jakkrapong Sangchoat*, Thidarat Koomsang, Chonnapat Naktubtim, Witchuda Payuhakrit

This research aimed to study of the optimum time of collagen staining method in fibroblast cells by using Masson trichrome with aniline blue dye staining kit. The V79 (Chinese hamster lung fibroblast) and CCD18-co ( human colon fibroblast) were used in this study. The fibroblast cells were cultured in 6 - well plates and then stained collagen in the fibroblast cells followed the manufacturer’s instructions, Masson’s trichrome with aniline blue (Bio-Optica, Italy) and modified method with adjusted times for staining. In the modified technique, the staining time of Masson aniline blue staining was adjusted from 5 to 10, 20, and, 30 minutes. The results
showed the standard method at 5 minutes staining of Masson aniline blue was unable to demonstrate the collagen in both V79 and CCD18-co cells by showing the pink color in cytoplasm. When increased staining time, the modified method at 1 0 and 2 0 minutes showed the dark red and purple color of collagen staining, respectively and while the modified method at 30 minutes showed clearly of the collagen content in dark blue color in both cell lines. The results showed the modified Masson trichrome with aniline blue staining at 30 minutes was developed in order to assist in demonstrated intracellular collagen. This modified method can be
used basically to support both study and research related to the study of collagen synthesis in fibroblast cells.