In corporation with Prof. Bruno Weber we custom-built a highly versatile multiphoton microscope. The microscope was financed by the UZH and NCCR Kidney to foster the in vivo imaging of kidney function and diseases (NCCR Kidney: Prof. Andrew Hall) as well as other research topics. The microscope allows full 360 degrees rotational freedom of the objective and large x, y, z movements. It is equipped with an advanced laser system with a very broad tuning range allowing deep tissue imaging and the excitation of red fluorescent dyes, which previously were not usable in multiphoton imaging.
Light Sources and Lasers
- Spectra-Physics InSight DeepSee Dual Ultrafast Ti:Sapphire laser system. Tunable between 680-1300 nm. Typical pulse with is 120 fs and the repetition rate is 80 MHz. Laser powers at different wavelengths: @680 nm: 780 mW, @900 nm: 1440 mW, @1000 nm: 1300 mW, @1200 nm: 1100 mW, @1300 nm: 950 mW, second output @1040: 685 mW. A dispersion compensation unit allows optimal pulse shaping for deep tissue imaging (DeepSee)
- Modulated green (530 nm) LED (Thorlabs M530L2) with custom built condenser for direct sample illumination
- Modulated white LED (Thorlabs MWWHL3) with custom built condenser for transmission illumination
|Olympus XLPlan N ( XLPLN25XWMP2)||25x||1.05||Water||2.0|
|Zeiss W Plan-Apochromat||20x||1.0||Water||2.3|
To mount microscope objectives from different manufacturers, the following objective adapters are available:
|M25x0.75||For Olympus XLPlan N and other objectives|
|M27x0.75||For Zeiss W Plan-Apochromat and other objectives|
|26x0.706||For Olympus objectives|
|0.8”x1/36”||RMS (Zeiss and Olympus objectives)|
|Name||Dichroic||IR Block||Detector 1||Detector 2|
|Cube 1||506 nm||680 nm SP||475/50 BP||542/50 BP|
|Cube 2||560 nm||770 nm SP||520/70 BP||607/70 BP|
|Cube 3||580 nm||770 nm SP||535/50 BP||630/69 BP|
IDS 1540LE CMOS monochrome camera, 25 fps, 1280x1024 pixels. The camera is used for aligning the instrument and for finding the focus. Fluorescent imaging is not possible.
- Custom made, heated mouse holder for inverted in-vivo experiments
- Anesthesia system for small rodents
- Binocular for surgery of small animals
- Two custom made microscope stages. An upright stage with transmission LED for imaging microscope slides and tissue slices in perfusion chambers. An inverted Stage for in-vivo experiments
- Micromanipulator for physiology experiments
Make sure to acknowledge the Center for Microscopy in your publication to support us.
- Laser safety is a very important aspect in operating custom built two-photon microscopes. The infrared laser used in this system is a class 4 laser. To reduce the risk of injury various technical and organizational measures have been taken. Therefore, access to the microscope is highly restricted and only well trained and experienced users are allowed to operate the system. It is compulsory to understand and accept laser safety measures and rules by signing the laser safety regulations before access to the system is given.
- The microscope containment is completely built using industrial aluminum profiles. This allows easy adaptions and expansion according to experimental needs.
Literature and Links