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IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Jayangshu Sengupta Cornea Services, Priyamvada Birla Aravind Eye Hospital Kolkata, India Nibaran Gangopadhyay Head-Eye Care, Sanjiban Hospital, Howrah, India Merle Fernandez Director & Cornea Consultant, Vizag-LVPEI,AP, India The authors have no financial interest in the subject matter of the poster IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Conjunctival impression cytology Simple, noninvasive technique Sample the superficial layers of epithelium for analysis. Gold standard procedure for analysis of squamous metaplasia and goblet cell loss. Light microscopy- remains most commonly used in clinical practice. Superior Limbic Keratoconjunctivitis Disorder of unknown etiology Role of impression cytology as a diagnostic modality remains largely unknown. IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID To demonstrate the morphological changes induced in the conjunctival epithelium in presence of SLK using the process of impression cytology. Prospective, non-comparative, observational case series 50 eyes(26 patients) were analyzed 2 eyes with inadequate samples were not analyzed January 2008 – December 2008 IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Diagnostic criteria Localized superior bulbar congestion with conjunctivochalasis +/- filament Localized punctuate Rose Bengal staining Papillary changes over upper tarsal conjunctiva Bilateral involvement. Evaluation Detailed history Slit lamp examination Schirmer test Tear film break up time Rose Bengal staining Fluorescein stain score Systemic examination Exclusion criteria Use of contact lens Schirmers II value<5mm Prior steroid therapytopical/systemic (within previous 3 months) IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Standard technique of impression cytology Haematoxylin- PAS staining Grossly labeled as normal/abnormal epithelial cell morphology cohesion of cells keratinization nuclear characteristics goblet cell density. Abnormal specimens graded Tsengs classification system Goblet cells Normal epithelial pattern (Scheffer C.G.Tseng Ophthalmology 92:728-733,1985) Abnormal Impression IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Age Range was found to be 36 to 55 yrs M:F ratio of 1:12 Co morbidity factors were found in 4 patients Hypothyroid Rheumatoid Arthritis 1 patient each Post Lasik High Prolactin level All samples from superior bulbar portion of conjunctiva demonstrated Grade V squamous metaplasia IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Increase in cell size Lack of cohesiveness with increased intercellular spaces and folded edges Keratinisation Complete absence of goblet cells Pyknotic nucleus IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Snake chromatin 100% Snake like chromatin Due to alteration in the nuclear and cytoplasmic skeleton Knop E, Reale E Fine structure and significance of snake like chromatin in conjunctival epithelial cells. Invest. Ophthalmol. Visual Sci 35:711, 1994 Margarita Calonge, Yolanda Diebold, Victoria Saez et al Impression cytology of the ocular surface: a review Experimental Eye Research78:457, 2004 IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Multinucleation (100 perent) Balloon degeneration Multinucleation Balloon degeneration (96%) IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Multilobulated nuclei (70%), Multilobulation Spindle Configuration Spindle configuration (68%) Dumb-bell configuration (60%). Dumb- bell Dumb- bell IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID Hallmark of squamous metaplasia from histopathological specimens Extensive nuclear pyknosis Condensation of masses of chromatin The shrinkage of the nuclear envelope from its surrounding cytoplasm Though electron microscopy of histological samples in patients with SLK better demonstrates such nuclear changes, conjunctival impression cytology in SLK conclusively demonstrates Localized squamous metaplasia of high grade Complete lack of goblet cells Characteristic nuclear changes like balloon degeneration, condensed chromatin, snake like chromatin, multi lobulated and strangulated nuclei Formation of snake like chromatin is characteristic Probably an indicator of mechanical stress on the ocular surface like blink related micro-trauma. Barry Collin, Peter C. Donshik, S. Arthur Boruchoff, et al. The fine structure of nuclear changes in superior limbic keratoconjunctivitis. Invest. Ophthalmol. Visual Sci 17(1):79, 1978 IMPRESSION CYTOLOGIC ANALYSIS OF SUPERIOR LIMBIC KERATOCONJUNCTIVITIS: NEW DIAGNOSTIC AID To conclude, impression cytology is a simple, noninvasive method of demonstrating the underlying pathological changes and thereby becomes an important tool in diagnosing SLK.