Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube
- Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States)
- Department of Mechanical Engineering, Dankook University (Korea, Republic of)
- Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States)
Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.
- OSTI ID:
- 22584032
- Journal Information:
- AIP Advances, Vol. 5, Issue 8; Other Information: (c) 2015 Author(s); Country of input: International Atomic Energy Agency (IAEA); ISSN 2158-3226
- Country of Publication:
- United States
- Language:
- English
Similar Records
Signal readout from scintillating fiber by using multianode photomultiplier tubes
Characterisation of signal-induced noise in Hamamatsu R11265 Multianode Photomultiplier Tubes