Two-photon excitation fluorescence (TPEF) microscopy imaging was performed on mRIC facility of Biosit, University of Rennes1 (France). The imaging system composed of a confocal TCS SP5 scanning head (Leica Microsystems, Mannheim, Germany) was mounted on a inverted microscope Leica DMSI 6000 CS (Leica Microsystems) and equipped with a MAITAI femtosecond laser (Spectra Physics, Santa Clara, CA). A 10× dry objective (NA = 0.4; Leica Microsystems), a 20× oil immersion objective (NA = 0.7; Leica Microsystems) and 60× water immersion objective (Olympus LUMFL 60 W × 1.1NA) were used. Image processing was performed using ImageJ software (National Institutes of Health; http://rsb.info.nih.gov.gate2.inist.fr/ij/ ).
Tcs sp5 scanning head
Manufactured by Leica Microsystems
The Leica TCS SP5 scanning head is a confocal laser scanning microscope component that provides high-resolution imaging capabilities. It utilizes a multi-channel detection system to capture fluorescence signals from samples. The scanning head is designed to be integrated with other Leica Microsystems imaging solutions.
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2 protocols using tcs sp5 scanning head
1
Two-Photon Excitation Fluorescence Microscopy
2
Multiphoton Microscopy of Biological Samples
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Imaging with TPEF microscopy was performed at the Mric facility of Biosit, University of Rennes1 (Fr). The TPEF imaging system is composed of a confocal TCS SP5 scanning head (Leica Microsystems, Mannheim, Germany), which is mounted on a DMIRE2 inverted microscope (Leica Microsystems). It was equipped with a Multiphoton Ma Ta HP Ti: Sapphire Mode Locked Laser (Spectra Physics, Santa Clara, CA) used to excite the samples at 810 nm. A 20x oil immersion and a 60x water immersion objective (Olympus LUMFL 60W x 1.1NA) were used. The TPEF was epi-collected in the backward direction. IRSP 715 bandpass and 405 nm infrared (IR) filters (10 nm full width at half-maximum, FWHM) were placed before the photomultiplier tube. Image processing was performed with ImageJ software (National Institutes of Health, (http://imagej.nih. Gov/ij/)).
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