Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/200026
Type: Artigo de periódico
Title: Harmonic Optical Microscopy And Fluorescence Lifetime Imaging Platform For Multimodal Imaging.
Author: Pelegati, Vitor B
Adur, Javier
De Thomaz, André A
Almeida, Diogo B
Baratti, Mariana O
Andrade, Liliana A L A
Bottcher-Luiz, Fátima
Cesar, Carlos L
Abstract: In this work, we proposed and built a multimodal optical setup that extends a commercially available confocal microscope (Olympus VF300) to include nonlinear second harmonic generation (SHG) and third harmonic generation (THG) optical (NLO) microscopy and fluorescence lifetime imaging microscopy (FLIM). We explored all the flexibility offered by this commercial confocal microscope to include the nonlinear microscopy capabilities. The setup allows image acquisition with confocal, brightfield, NLO/multiphoton and FLIM imaging. Simultaneously, two-photon excited fluorescence (TPEF) and SHG are well established in the biomedical imaging area, because one can use the same ultrafast laser and detectors set to acquire both signals simultaneously. Because the integration with FLIM requires a separated modulus, there are fewer reports of TPEF+SHG+FLIM in the literature. The lack of reports of a TPEF+SHG+THG+FLIM system is mainly due to difficulties with THG because the present NLO laser sources generate THG in an UV wavelength range incompatible with microscope optics. In this article, we report the development of an easy-to-operate platform capable to perform two-photon fluorescence (TPFE), SHG, THG, and FLIM using a single 80 MHz femtosecond Ti:sapphire laser source. We described the modifications over the confocal system necessary to implement this integration and verified the presence of SHG and THG signals by several physical evidences. Finally, we demonstrated the use of this integrated system by acquiring images of vegetables and epithelial cancer biological samples.
Subject: Adenocarcinoma, Mucinous
Female
Humans
Image Processing, Computer-assisted
Microscopy, Confocal
Microscopy, Fluorescence
Onions
Ovarian Neoplasms
Solanum Tuberosum
Rights: fechado
Identifier DOI: 10.1002/jemt.22078
Address: http://www.ncbi.nlm.nih.gov/pubmed/22648907
Date Issue: 2012
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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