| Texto completo | |
| Autor(es): |
Pelegati, Vitor B.
[1, 2]
;
Adur, Javier
[1, 2]
;
de Thomaz, Andre A.
[1]
;
Almeida, Diogo B.
[1]
;
Baratti, Mariana O.
[1]
;
Andrade, Liliana A. L. A.
[3]
;
Bottcher-Luiz, Fatima
[4]
;
Cesar, Carlos L.
[1]
Número total de Autores: 8
|
| Afiliação do(s) autor(es): | [1] State Univ Campinas UNICAMP, Biomed Lasers Applicat Lab, Opt & Photon Res Ctr, Dept Eletron Quont, Gleb Wataghin Inst Phys, BR-13083859 Sao Paulo - Brazil
[2] Natl Univ Entre Rios UNER, Sch Bioengn, Microscopy Lab Appl Mol & Cellular Studies, RA-3101 Oro Verde, Entre Rios - Argentina
[3] State Univ Campinas UNICAMP, Dept Pathol, Fac Ciencias Med, Dept Anat Patol, BR-13083970 Sao Paulo - Brazil
[4] State Univ Campinas UNICAMP, Dept Obstet & Gynecol, Fac Ciencias Med, Lab Citogenet & Cult Celular, Dept Tocoginecol, CAI, BR-13083970 Sao Paulo - Brazil
Número total de Afiliações: 4
|
| Tipo de documento: | Artigo Científico |
| Fonte: | MICROSCOPY RESEARCH AND TECHNIQUE; v. 75, n. 10, p. 1383-1394, OCT 2012. |
| Citações Web of Science: | 12 |
| Resumo | |
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. Microsc. Res. Tech. 2012. (c) 2012 Wiley Periodicals, Inc. (AU) | |