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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| 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. |
| Description | Author Affiliation: Pelegati VB ( Optics and Photonics Research Center, Gleb Wataghin Institute of Physics, State University of Campinas (UNICAMP), Departamento de Eletrônica Quôntica, Campinas, São Paulo. Brazil.) |
| 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. |
| File Format | HTM / HTML |
| ISSN | 1059910X |
| Issue Number | 10 |
| Volume Number | 75 |
| e-ISSN | 10970029 |
| Journal | Microscopy Research and Technique |
| Language | English |
| Publisher | Wiley-Liss |
| Publisher Date | 2012-10-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Chemistry Techniques Microscopy, Confocal Methods Microscopy, Fluorescence Adenocarcinoma, Mucinous Pathology Female Humans Image Processing, Computer-assisted Onions Cytology Ovarian Neoplasms Solanum Tuberosum Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Anatomy Histology Instrumentation Medical Laboratory Technology |
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