Building a 2-photon microscope

Less than a year ago, the patent on 2-photon microscopy ran expired (Nov 14, 2009 pdf). So now anyone can sell a kit or system for two photon microscopy without having to pay a licensing fee.

If you don’t want to start from scratch, there are a lot of options.

Prairie Technologies

Prairie Technologies – Prairie started out collaborating with Svoboda on designs and using ScanImage, but now they have a lot of new products and have written their own software. Although their code is closed, Prairie offers very quick turnaround for interfacing with their software. So if you want to change something in the code, you can talk to the programmers and find a way to do what you want.

LaVision BioTec

LaVision BioTec is best known for the 64-beam splitting systems with high speed CCD aquisition. While this offers great speed, the imaging at depths below 50 microns is not as good as imaging with a traditional single beam + PMT approach. They also sell regular 2p scopes, including self-aligning systems.

Femtonics

Femtonics is a Hungarian company in Budapest. They sell 2D and 3D 2p systems, accessories, and caged compounds.

Sutter Instruments

Sutter Instruments sells the MOM (moving objective microscope), aka the Denkscope. This very compact design from Winfried Denk himself, places the objective on an x-y-z manipulator, leaving the prep to remain stable. In addition, the assembly can be rotated in the z-x plane. Finally, the entire scope can be easily slid away from the experimenter, to permit a dissecting scope to be positioned for the surgery. The detection pathway must be kept to a low mass, in order to allow the manipulator to function, but there are several different configurations available.

Scientifica

Scientifica has a barebones microscope platform upon which a 2p system can be built. Instead of writing their own software, they are working with the ScanImage group to make their stage compatible (ScanImage was developed for MP-285 based systems). This is an interesting system that is somewhere between turn-key and custom. It could be a great place to start for experimenters who want to build their own but not start from scratch.

Other options

Newport had an opportunity to put together a kit, but blew it. Maybe they’ll try again. The sell almost all the needed parts including the laser, they just need someone knowledgable to put together a list of SKUs.

The major microscope manufacturers (Olympus, Nikon, Leica, and Zeiss) will help you build a 2p system off of one of their scopes. For example, they might cut a hole in the side to allow you to mount non-descanned detectors in the optical pathway. They won’t build the whole system for you, of course, but they can offer a few tips and modifications to make it easier. And it goes without saying, they’ll be more than happy to sell you their turn-key systems as well (e.g., the Olympus FV1000 MPE and the Zeiss LSM 710 NLO).

I’ve built 2p rigs from scratch before. It’s really not that difficult, but it can be time consuming. Particularly if you don’t have experience with building or don’t have access to a machine shop with fast turnaround. However, there are a lot of papers and resources to help if you’d like to go that route. The Tsai book chapter (first edition) is a great one to start with. The main benefit of building from scratch is that you can optimize the optics and mechanical design to fit your planned experiments exactly.

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