The Research On The Biological Properties Of Malassezia And Other Fungal Autofluorescence By Laser Scanning Confocal Microscope

Objective:①To study the autofluorescence characteristics of the pathogenicfungi, such as Malassezia spp., Candida spp., under ultraviolet radiation.②To detect the rule of the quantity and quality of autofluorescence when thefungi were in acid or alkaline circumstance, living or death condition.③Toestablish the optimum condition of detecting and recording autofluorescenceof fungi.④To explore the possibility of early and rapid diagnosis of mycosisby detecting the fungal autofluorescence.Methods: To culture M. furfur and M. japonica standard strains on Dixonmedium at 32℃, and to study the autofluorescence of colonies in 2nd, 4th, 6th, 8th, 10th day under Laser Scanning Confocal Microscopy (LeicaTCS-SP2 LSCM). To study the variation of autofluorescence whenMalassezia colonies were put in 3% glutaraldehyde, pH4.0 HCL, 10% KOH, fluconazol solution, 0.3% methylene blue separately. To detect theautofluorescence of C. albicans, S. schenchii, T. rubrum, T. tonsurans, A.terreus, A. fumigatus cultured on Sabouraud dextrose agar (SDA). Results: The autofluorescence of M. furfur and M. japonica standard trainsrespectively in 4th and 6th day are stronger than that in other days, andautofluorescence of colonies become weaker in an hour. Theautofluorescence of Malassezia colonies put in 3% glutaraldehyde, pH4.0HCL, 1280μg/mL and 64μg/mL fluconazol solution, 0.3% methylene blueseparately get stronger, and that in 10% KOH get stronger at beginning thenbecome weaker 2 days later. C. albicans, S. schenchii, T. rubrum, T.tonsurans, A. terreus, A. fumigatus cultured on SDA all haveautofluorescence, and which are weaker than that of Malassezia.Conclution:①The autofluorescence of different species and strains aredifference.②The autofluorescence of the same strain cultured on the samemedium are difference at different time.③Both weak acid and alkalinecircumstance can enhence the autofluorescence of Malassezia, and 10%KOH makes that become stronger at beginning then become weaker 2 dayslater.④Either in living condition or death the autofluorescence of fungi isexist.⑤C. albicans, S. schenchii, T. rubrum, T. tonsurans, A. terreus, A.fumigatus cultured on SDA all have autofluorescence, and which are weakerthan that of Malassezia.⑥The autofluorescence of spores are stronger thanthat of hypha.