Application of confocal laser scanning microscopy for the diagnosis of amyloidosis
| Autor: | Mila Della Barbera, Gaetano Thiene, Fausto Adami, Chiara Castellani, Annalisa Angelini, Marialuisa Valente, Anna Chiara Frigo, Marny Fedrigo |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male Amyloid Pathology medicine.medical_specialty Histology Materials science Confocal Thioflavin-T Sensitivity and Specificity Pathology and Forensic Medicine 03 medical and health sciences chemistry.chemical_compound Microscopy Electron Transmission Microscopy Confocal laser microscopy Fluorescence microscope medicine Humans Coloring Agents Molecular Biology Microscopy Confocal Amyloidosis fungi Resolution (electron density) Congo Red Cell Biology General Medicine medicine.disease Congo red 030104 developmental biology chemistry Congo red fluorescence Female Amyloid (mycology) |
| Zdroj: | Virchows Archiv : an international journal of pathology. 470(4) |
| ISSN: | 1432-2307 |
| Popis: | We analysed specificity and sensitivity of confocal laser microscopy (CLSM) on tissue sections for a diagnosis of amyloidosis, in an attempt to reduce technical errors and better standardise pathological diagnosis. We first set up a protocol for the use of CLSM on this type of specimen, using a group of 20 amyloid negative and 20 positive samples. Of all specimens, 2, 4 and 8-μm sections were cut. Sections were stained with Congo red (CR) and thioflavin-T (ThT) and observed by cross-polarised light microscopy (CR-PL), epifluorescence microscopy (CRF-epiFM and ThT-epiFM) and CLSM (CRF-CLSM and ThT-CLSM). To validate the method in a diagnostic setting, we examined tissue samples from 116 consecutive patients with clinical suspicion of amyloidosis, selected from the period 2005 to 2014 from the database of the Pathology Unit of the University of Padua. The results were compared with those of transmission electron microscopy (TEM), which we consider as reference. We found that with CRF-CLSM, the false negative rate was reduced from 17 to 5%, while the sensitivity of detection increased to 12%. The results were in complete agreement with those of TEM ThT-CLSM; both sensitivity and specificity were 100%. Finally, ThT-CLSM results did not vary with section thickness, and small amounts of amyloid could even be detected in 2-μm sections. In conclusion, we found ThT-CLSM to be more sensitive as a screening method for amyloidosis than CR and ThT epifluorescence optical imaging. The method was easier to standardise, provided images with better resolution and resulted in more consistent pathologist diagnoses. |
| Databáze: | OpenAIRE |
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