The Application Study Of Confocal Microscopy Combined With Dark Field Microscopy In The Fungal Keratitis

Objective1. To study the cellular morphology of fungal keratitis and culture colony smears by confocal microscopy in vivo. Then assess the diagnosis effect of confocal microscopy in the fungal keratitis;2. To study the cellular morphology of fungal keratitis by confocal microscopy in vivo before and after the corneal scraping, then get the best inspection time;3. The patients with atypical fungal keratitis (can not be diagnosed the fungal keratitis by regular corneal scraping, smearing and microscopy) were observed by dark field microscopy and in vivo confocal microscopy. The clinical characteristics are summarized. To assess the values of dark field microscopy in early diagnosis of fungal keratitis;4. To improve the level of diagnosis of fungal keratitis according to the the results above.Methods1.34 cases of typical fungal keratitis and culture colony smears were examined by by laser confocal microscopy in vivo. Images were recorded by continuously focusing on the optical section through the corneal ulcer. The images were analyzed visually for morphology and compared with each other by Heidelberg Tomograph III Cornea Module.2. Images were obtained from 45 cases of typical fungal keratitis were examined by confocal microscopy in vivo before and after the corneal scraping. To study the cellular morphology of the fungal keratitis by the images which we got. Find out the best inspection time of the confocal microscopy.3.23 cases (23 eyes) of atypical fungal keratitis were examined by in vivo confocal microscopy, then scraped the corneal ulcer and smeard,were examined by regular bright field microscopy and Dark field microscopy. The characteristics of their images were studied and compared.Results1. The cellular morphology of hyphae and spores by in vivo confocal microscopy in fungal keratitis are the same as that with culture colony smears basically.2. The cellular morphology of hyphae with confocal microscopy in fungal keratitis is intactand clear before the corneal scraping. After the corneal scraping, the cellular morphology of hyphae with confocal microscopy in fungal corneal ulcer get broken and shadowy.3. The fungal hyphae of atypical fungal keratitis in vivo confocal microscopy was not complete, and the imagings were not clear. The images of fungal hyphae were unclear or difficult to diagnose in regular bright field microscopy. However, the clear fungal hyphae can be found by dark field microscopy.Conclusions1. The diagnostic results of confocal miscrocopy in fungal keratitis in vivo are basically the same as that of culture colony smears. The in vivo confocal miscrocopy is an useful method in diagnostic studies of fungal keratitis.2. We should take the examination of the in vivo confocal microscopy before the corneal scraping to advance the diagnostic level in the fungal keratitis in the clinical.3. The dark field microscopy is an useful method in the diagnosis of typical fungal keratitis.4. There were unique advantage of dark field microscope that the fungal pyphae of atypical fungal corneal ulcer were observed. The dark field microscope, confocal microscopy and routine smear microscope can improve the diagnosis levels of atypical fungal keratitis.