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Fluorescence lifetime imaging microscopy (FLIM) applied in lung exsudates

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Author(s):
Ana Paula Racanelli Bovo
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas
Defense date:
Examining board members:
Konradin Metze; Albina Messias de Almeida Milani Altemani; Ruy Gastaldoni Jaeger
Advisor: Konradin Metze
Abstract

Pathology is mainly based on methods introduced in the second half of the 19th century. New technologies could be very helpful for making diagnosis. Fluorescence lifetime imaging (FLIM) is a new technique, where the contrast in the fluorescence image is based on the lifetime of the fluorophores. We used paraffin blocks from the files of surgical and autopsy pathology of the department of Pathology, FCM/UNICAMP. 53 cases with pulmonary eosinophilic transudates or exsudates were randomly selected from the time span between 1966 and 2014. Slightly hematoxylin stained paraffin sections were examined with the help of a confocal microscope with a FLIM equipment. In order to validate the method we analyzed first small arteries. The lifetimes between erythrocytes, smooth muscle , collagen and elastic fibres were different in each of the cases and we were able differentiate these structures in the virtual images by pseudocolors, although the lifetime values of a certain structure varied considerably between the cases, we were able to create reproducible virtual images by adjustment of the pseudocolors. Regarding eosinophilic exsudates and transudates, the lifetime of the alveolar proteinosis cases was significantly longer than those of the other groups. Tests of randomness showed that the texture of the edema and proteinosis were significantly different from that of the pneumonias or hyaline membranes. The distribution of the lifetime values across a hyaline membrane could be approximated by a parabolic curve. Our study suggests that FLIM can reveal valuable complementary information for the examination of lung specimens (AU)

FAPESP's process: 13/18468-9 - Differentiation of exudates with new methods of fluorescence microscopy
Grantee:Ana Paula Racanelli
Support Opportunities: Scholarships in Brazil - Master