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Updated: Aug 9, 2026
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Primary intraocular lymphoma: clinical, cytologic, and flow cytometric analysis
Renzo A Zaldivar1, Daniel F Martin, Jeannine T Holden
1Department of Ophthalmology, Emory University, Atlanta, Georgia, USA.
Insights
Cytology accurately diagnoses primary intraocular lymphoma (PIOL). Flow cytometric immunophenotyping (FCI) aids in PIOL immunophenotyping, though multiple biopsies may be needed for diagnosis.
Area of Science:
- Ophthalmology
- Hematology
- Oncology
Background:
- Primary intraocular lymphoma (PIOL) is a rare non-Hodgkin lymphoma.
- Diagnosis often relies on vitreous biopsy analysis.
- Comparing diagnostic methods is crucial for accurate PIOL detection.
Observation:
- Vitreous biopsy specimens were analyzed using both cytology and flow cytometric immunophenotyping (FCI).
- Both methods identified chronic inflammation and large cell lymphoma in most cases.
- Cytology diagnosed lymphoma in one case initially negative by FCI.
Findings:
- Cytology demonstrated high accuracy in diagnosing PIOL.
- FCI proved valuable for immunophenotyping identified lymphomas.
- Sufficient specimens for FCI were not always obtained, highlighting potential limitations.
Implications:
- Cytologic evaluation is a reliable method for PIOL diagnosis.
- FCI complements cytology by providing immunophenotypic details.
- Consideration of repeat biopsies may be necessary for definitive PIOL diagnosis.
Purpose:
To compare cytologic with flow cytometric results of vitreous biopsy specimens obtained to rule out primary intraocular lymphoma (PIOL).
Study Design:
Prospective noncomparative case series.
Participants:
Patients suspected of having PIOL who underwent vitreous biopsy were evaluated.
Methods:
Patients underwent a standard 3-port vitrectomy and vitreous biopsy to rule out PIOL. Each undiluted specimen was split, and half was prepared for cytologic evaluation with the collodion bag method; the other half was submitted for flow cytometric immunophenotyping (FCI). The diluted specimen was processed as a cell block for cytology.
Main Outcome Measures:
Final diagnosis based on cytology and FCI.
Results:
Ten of 14 patients had sufficient specimens for both cytologic and FCI evaluation. Three patients had chronic inflammation confirmed by both methods. Six patients had large cell lymphoma identified by both cytology and FCI. Two of those 6 patients initially had insufficient specimen for FCI. One patient had large cell lymphoma diagnosed cytologically that was initially negative for a clonal population by FCI. All lymphomas were B-cell type.
Conclusions:
Cytologic evaluation is an accurate diagnostic technique to evaluate for PIOL. FCI is useful for immunophenotyping PIOL. Multiple biopsies may be required to achieve a diagnosis.
