Cytopathological analysis of vitreous in intraocular lymphoma

Abstract

Objective To describe the cytopathological method used in the analysis of vitreous samples in the diagnosis of primary intraocular lymphoma (PIOL). Participants Seven patients with refractory posterior uveitis referred to a regional ocular inflammatory service were diagnosed as having PIOL between 1999 and 2006. Methods Clinical features of the uveitis and cytopathological preparation of the samples were described. All patients underwent vitrectomy and samples were placed in formal saline or prepared fresh. Following paraffin embedding generating a cell block, immunostaining, and polymerase chain reactions were performed. Results Five women (71.4%) and two men (28.6%) (mean age 67.7 years) were included. Five patients had diagnostic vitrectomy performed within 6 months of presentation, but in two patients diagnosis was delayed up to 2 years. Uveitis was bilateral in two patients. Cytologic and immunohistochemical staining prepared from the vitreous specimens showed PIOL in all patients, and PCR displayed single band of immunoglobulin heavy chain rearrangement in five out of six samples tested. Conclusions Diagnosis of PIOL is difficult due to small volume of sample with low number of malignant cells and inadequate preparation of samples. Our method of analysis with fresh samples together with immunohistochemistry and PCR analysis demonstrates a high yield of diagnosis reducing diagnostic delay.

The pathologist's perspective on vitreous opacities

“…[4][5][6][7]9,10,20,21,26,[30][31][32][33][34][35] These include: (1) vitreous 'filtration'; (2) a celloidin bag technique; (3) cytospin and (4) cell block preparation. We are most familiar with the latter two methods, whereby the vitreous is spun at 500 r.p.m.…”

Current concepts and future directions in the pathogenesis and treatment of non-infectious intraocular inflammation

“…Nevertheless, of equal importance is the exclusion of infection or masquerade, and AC sampling is increasingly being used for this purpose, the most classic example of which is the advances achieved in the diagnosis of primary intraocular lymphoma by measurement of aqueous IL-10 and/or IL-10: IL-6 ratios, and by vitreous biopsy, utilising modern immunohistochemistry, flow cytometry, and PCR to facilitate diagnosis. 103 The utility of sampling AC or vitreous is also to discriminate infectious causes from autoimmune/inflammatory disease. By PCR or by Goldmann-Witmer co-efficient of specific antibody production against microbes/viruses we are now able to discern more precisely possible infectious causes, which as a result will direct treatment appropriately.…”

An Update on Tubercular Uveitis

“…The preparation of vitreous samples for cyto-pathological analysis has also changed over time to improve yields from this limited specimen. Intzedy et al reported that samples placed in saline or prepared fresh followed by paraffin embedding were able to yield a positive diagnosis [48].…”