A multiphoton laser scanning microscope setup for transcranial in vivo brain imaging on mice

Nase, Gabriele; Helm, P. Johannes; Reppen, Trond; Ottersen, Ole Petter
December 2005
Review of Scientific Instruments;Dec2005, Vol. 76 Issue 12, p123702
Academic Journal
We describe a multiphoton laser scanning microscope setup for transcranial in vivo brain imaging in mice. The modular system is based on a modified industrial standard Confocal Scanning Laser Microscope (CSLM) and is assembled mainly from commercially available components. A special multifunctional stage, which is optimized for both laser scanning microscopic observation and preparative animal surgery, has been developed and built. The detection unit includes a highly efficient photomultiplier tube installed in a Peltier-cooled thermal box shielding the detector from changes in room temperature and from distortions caused by external electromagnetic fields. The images are recorded using a 12-bit analog–to–digital converter. Depending on the characteristics of the staining, individual nerve cells can be imaged down to at least 100 μm below the intact cranium and down to at least 200 μm below the opened cranium.


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