Confocal laser scanning microscopy를 이용한 표피내 칼슘기울기 관찰

Abstract

Background: Ion capture cytochemistry(the potassium oxalate pyroantimonate method), semiquantitatively, and proton probe X-ray microanalysis, quantitatively, have been applied to investigate the epidermal calcium distribution. Objective:In this study, we examined the epidermal calcium distribution with confocal laser scanning microscopy(CLSM) in an attempt to evaluate the possibility of another method in epidermal calcium study. Methods:The change of epidermal calcium distribution after barrier perturbation with tape stripping was investigated with CLSM and was compared to the results of ion capture cytochemistry. Results:The calcium distribution pattern in normal murine epidermis demonstrated by CLSM show a normal calcium gradient, from a low level of calcium ions in the basal and spinous layer, followed by a progressive increase with a level of calcium ions reaching its maximal density within the outer stratum granulosum. Disruption of the epidermal barrier with tape stripping induced an immediate loss of the calcium gradient and the calcium gradient after 36h was almost normalized, in parallel with the recovery of barrier function. Conclusion:These results show that calcium gradient in murine epidermis after tape-stripping is restored by 36h and CLSM study can be used as a new method in epidermal calcium study.