Imaging, Manipulation and Optogenetics in Zebrafish by Itia Amandine Favre‐Bulle

Author:Itia Amandine Favre‐Bulle
Language: eng
Format: epub, pdf
ISBN: 9783319962504
Publisher: Springer International Publishing

SLM for light sensitive cells targeting. Once again, the transparency of the brain and the absence of pigments allow to reach the brain more easily. Additionally the flexibility of SLM will allow to drive single or multiple cells in 3D.

The system uses the same configuration of 488 nm laser and SLM presented in Chap. 3 and Fig. 3.​3, the upgrade being a different microscope objective (with a higher NA and longer working distance). In addition to the SLM illumination and imaging system we added two SPIM planes orthogonal to each other and to the microscope objective. The configuration and combination of the optical system built are presented in Fig. 4.5.

Fig. 4.5Sketch of the set-up of the two SPIMs incorporated into the previous set-up. a Image of the overall configuration. b Chamber model designed with Tinkercad and latter printed on a 3D printer. 20 20 mm coverslips were glued on each side of the chamber for minimal SPIM distortions. c Details of the two SPIMS set-ups. For illumination, we used a 150 mW, 488 nm OBIS laser coupled to an optical fibre. The laser light was expanded and collimated A 50/50 splitter splits the beam into two beam with equal intensity. Each beam passes through a horizontal slit, a cylindrical lens and a 4, 0.28 NA Olympus microscope objective. The two light sheets emerging from the microscope objectives focus down into the sample chamber where the zebrafish is set

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