The Autofluorescence Of Eccrine Gland In Skin Tissue And Analysis Of Confocal λ Scanning

Background:100 years ago,Stubelrepoted the potential use of cell and tissue autofluorescence for diagnostic purpose.In 1924,Policard reported a study concerning the autofluorescence of tumours,attributed to endogenous porphrins.About 50 years ago,autofluorescence of eosinophil granulocytes and its use for diagnostic purpose was studied.Due to the low intensity and spectral complexity of autofluorescence,further research has been limited.More recently,the availability of high-sensitivity,low signal-to-noise ratio device fluorescence microscope and confocal laser scanning microscope-CLSM,make the observation and measurement of autofluorescence become reality.Autofluorescence both in research and diagnostics have been reconsidered.Autofluorescence has discovered to correlate well with the diagnosis and treatment of some diseases including retinal disease、respiratory tumor and so on.Fluorophores in skin tissue causes skin autofluorescence,and some pathological process will also produce the endogenous and exogenous fluorophore.Advanced glycation end products can accumulate in the skin tissuecause skin autofluorescence has been proved important relationship with the atherosclerosis,cardiovascular disease,diabetes and its complications,kidney disease,hemodialysis and so on.There are few reports of eccrine sweat gland autofluorescence in the skin tissue.Our laboratory discovered autofluorescence during the extraction of eccrine sweat gland which has attracted our attention and induced a series studies of eccrine sweat gland optical character.Objective:We have known that monitoring skin autofluorescence can have both diagnostic and therapy significance in many diseases,but there is no such report about autofluorescence of eccrine sweat gland in skin.For the first time we found and describe the phenomenon of autofluorescence of eccrine gland during the process of this experiment,by detecting the autofluorescence characteristics and measuring autofluorescence intensity,it is very essential for eccrine gland autofluorescenceapply to related disease diagnosis and treatment.Methods:Haemat-oxylin-eosin(HE)stained skin tissue sections and observe eccrine sweat gland autofluorescence under a fluorescence microscope;Single eccrine sweat gland was extracted from the skin of volunteers and cultured in vitro,then we observe its autofluorescence under wide ultraviolet、wide green、wide blue of fluorescence microscope.Eccrine sweat glands autofluorescence changing with their growth is also observed;The autofluorescence intensity of the single eccrine sweat gland was measured using a laser confocal scanning microscope system(confocal laser scanning microscope,CLSM)to further understand the spectroscopy information of eccrine sweat glands and to predict the fluorophores according to it.Results: HE stained eccrine sweat glands autofluorescence in skin tissue sections shows green under GFP fliter system(470/40 nm),red under N2.1 fliter system(515–560 nm).Moerover,the single eccrine sweat gland showed three autofluorescencecolours under fluorescence microscope,including blue under wide ultraviolet、 green under wide blue and red under wide green.The autofluorescence intensity of the single eccrine sweat gland was showed two peaks around530 nm and 590 nm.The result may suggest flavin and lipopigment as the endogenous fluorophores.Conclusion: Monitoring HE stained eccrine sweat gland sections autofluorescence in skin tissue sections is helpful for easily distinguish the structure of eccrine sweat gland.Flavin and lipopigment maybe the fluorophores causes autofluorescence of the single eccrine sweat gland.