Improved detection of amyloid in fat pad aspiration: an evaluation of Congo red stain by fluorescent microscopy
Tamar A Giorgadze1, Natsuko Shiina, Zubair W Baloch
1Department of Pathology and Laboratory Medicine, Cytopathology and Cytometry Section, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, USA. tamar_giorgadze@yahoo.com
Abstract:
Amyloid fat pad aspiration specimens for cases with a clinical suspicion of amyloid typically are stained with Congo red and examined by brightfield microscopy. Congophilia with apple-green birefringence by polarization microscopy (PM) is considered diagnostic for amyloid. Examination of Congo red-stained slides by fluorescent microscopy (FM) is considered by some to be a more sensitive detection method. In this study, we assessed the utility of this technique in cytopathology archival slides from abdominal fat pad aspirations previously stained with Congo red dye. Seventy-eight cases of abdominal fat pad aspirations collected during the last 5 yr and stained with the Congo red procedure were obtained from archival files. Additionally, 20 adipose tissue material slides prepared from the surgical pathology specimens were examined as controls. One representative smear was examined in each case using FM equipped with rhodamine excitation/absorption (540/570 nm) filters. Relevant clinical information was obtained in all cases. Twelve cases (15.4%) of the 78 fat pad aspiration cases were reported originally as positive by Congo red stain using polarization and apple-green birefringence as diagnostic criteria. On review, four cases were deemed unsatisfactory. By FM examination 29 of the 74 (39.2%) cases were reclassified as positive for amyloid. The results were confirmed by immunohistochemical stain for amyloid P protein and electron microscopy. A number of similar distinct fluorescence and immunohistochemical patterns were recognized in the positive cases. Minimally weak fluorescence in the adipose tissue was observed in the control cases. The use of FM in Congo red-stained fat pad smears can improve the detection of amyloid in cytology preparations.
Insights
Fluorescent microscopy (FM) enhances amyloid detection in Congo red-stained abdominal fat pad aspirates. This sensitive method reclassified nearly 40% of initially negative cases as positive for amyloid.
Area of Science:
- Cytopathology
- Histopathology
- Medical Diagnostics
Background:
- Amyloidosis diagnosis typically relies on Congo red staining and polarization microscopy (PM).
- Fluorescent microscopy (FM) is proposed as a more sensitive method for amyloid detection.
Purpose of the Study:
- To evaluate the utility of FM for detecting amyloid in archival Congo red-stained abdominal fat pad aspiration slides.
Main Methods:
- Seventy-eight archival abdominal fat pad aspirates and 20 control adipose tissue slides stained with Congo red were examined using FM (rhodamine filters).
- Results were confirmed with immunohistochemical staining for amyloid P protein and electron microscopy.
Main Results:
- Original diagnosis: 12/78 (15.4%) positive for amyloid by PM.
- FM reclassified 29/74 (39.2%) cases as positive for amyloid.
- Distinct fluorescence and immunohistochemical patterns were observed in positive cases; minimal fluorescence in controls.
Conclusions:
- FM significantly improves amyloid detection in cytology preparations of abdominal fat pad aspirates.
- This technique offers enhanced sensitivity compared to traditional polarization microscopy for amyloid diagnosis.


