The Use of Multispectral Imaging in Fine Needle Aspiration Biopsies of Reactive Lymphoid Hyperplasia and Follicular Lymphoma
A Zarineh, J Yu, RE Felgar, WE Khalbuss, K Cieply, SE Monaco. University of Pittsburgh Medical Center, Pittsburgh, PA
Background: The cytologic distinction between reactive lymphoid hyperplasia (RLH) and small cell lymphomas, such as follicular lymphoma (FL), can be difficult in fine needle aspirations (FNA). The use of ancillary tests in these cases is crucial, and often includes flow cytometry (FC), immunohistochemistry (IHC), and fluorescence in-situ hybridization (FISH). The interpretation of IHC can be challenging due to the subjective and qualitative nature, in addition to the need for multiple sections for analysis of multiple antibodies. Multispectral imaging (MSI) with quantum dots involves highly fluorescent molecules that allow multiple spectra to be distinguished when used in combination, which allows for more precise signal quantitation. Thus, we examined the ability of MSI to provide quantitative and useful data to help in distinguishing RLH and FL in FNA biopsies.
Design: A total of 8 FNA cell block sections from 4 RLH and 4 FL cases were stained using immunohistochemical stains (CD3, CD20, CD10, and bcl-2) conjugated to quantum dot fluorophores. A series of slides were also used for controls and for creating a spectral library on the Nuance CRI Flex microscopy system (CRi, Cambridge Research & Instrumentation, Woburn, MA; spectral range 420 to 720 nm). A total of 3 fields of view (FOV) at 10x magnification were captured for each case and analyzed using the Nuance software version 2.10.
Results: All 8 cases were from FNAs of lymph nodes (4 FNA performed by pathologist, 2 CT-guided, 2 US-guided). The analysis of the 24 FOV revealed that the FL cases tended to show more CD20-positive cells than CD3-positive cells (lower CD3:CD20 ratio; 0.3 vs. 3.9), and tended to have more pixels colocalizing CD20/CD10/bcl-2. For quality control purposes, the average percent of pixels colocalizing CD3 and CD20 was less than 0.4%.
Conclusions: This study shows that MSI is feasible on FNA cell block material and can be helpful in obtaining quantitative data from multiple antibodies used on a single slide, which may be advantageous in FNA samples with limited material and may potentially be helpful in the cytologic distinction of RLH and FL.
Monday, March 22, 2010 9:30 AM
Poster Session I Stowell-Orbison/Surgical Pathology/Autopsy Awards Poster Session # 261, Monday Morning