Abstract
Diagnosis of intraocular lymphoma is difficult. Among the hurdles in the diagnosis are the variety of reactive inflammatory and ischemic changes among intraocular lymphoma patients. Thus, a novel diagnostic method is desired such that lymphoma cells can be distinguished by the signals intrinsic to the cells, not by those from the surrounding tissues with reactive changes. Raman spectroscopy is a technique that can detect intrinsic signals from each cell. Therefore, Raman spectroscopy is a good candidate for an intraocular evaluation technology that could contribute to improve the diagnosis of intraocular lymphoma. In this study, we tested whether the intrinsic Raman signals from malignant lymphoma cells, in the absence of surrounding tissue, were sufficient for the discrimination of malignant lymphoma cells from leukocytes. We acquired spectra from dissociated lymphoma cells, along with spectra from normal B cells and other leukocytes involved in intraocular inflammatory diseases. We analysed the spectra using principal component analyses and quadratic discriminant analyses. We found that Raman spectra from dissociated cells without confounding tissues showed high discriminating ability, regardless of the variation due to day-to-day differences and donor differences. The present study demonstrates the possible effectiveness of Raman spectroscopy as a tool for intraocular evaluation.
Highlights
Is poor, with median survival times of 58 m onths[14,15,16]
As a candidate technique for intraocular evaluation, we focused on Raman spectroscopy for its ability to optically detect the chemical and metabolic conditions of cell samples
Three Raman spectra were recorded in the phosphatebuffered saline (PBS) next to each cell (Fig. 2a)
Summary
Is poor, with median survival times of 58 m onths[14,15,16]. an early diagnosis is desirable, the actual diagnostic process often is time-consuming. Multiple cytology-based technologies have been employed to improve the diagnostic rate, including the cell block technique, intraocular fluid cytokine analysis, and PCR for detection of immunoglobulin gene rearrangement[15]. Raman spectroscopy previously has been reported to be useful for the diagnosis of malignant diseases, including malignant lymphoma[19,20,21] Those earlier reports evaluated lymph node and spleen sections that included surrounding cells and stroma, meaning that the acquired data could have been confounded by characteristics of the deteriorating surrounding tissues. It was not clear whether Raman signals would permit discrimination of malignant lymphoma from other diseases based on properties intrinsic to the lymphoma cells. We examined the potential effectiveness of Raman spectroscopy when applied for intraocular evaluation
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