Differential Multiphoton Laser Scanning Microscopy

Dept. of Phys., Colorado Sch. of Mines, Golden, CO, USA
IEEE Journal of Selected Topics in Quantum Electronics (Impact Factor: 3.47). 03/2012; 18(1):14 - 28. DOI: 10.1109/JSTQE.2010.2077622
Source: IEEE Xplore

ABSTRACT Multifocal multiphoton laser scanning microscopy (mfMPLSM) in the biological and medical sciences has the potential to become a ubiquitous tool for obtaining high-resolution images at video rates. While current implementations of mfMPLSM achieve very high frame rates, they are limited in their applicability to essentially those biological samples that exhibit little or no scattering. In this paper, we report on a method for mfMPLSM in which whole-field detection with a single detector, rather than detection with a matrix of detectors, such as a charge-coupled device (CCD) camera, is implemented. This advance makes mfMPLSM fully compatible for use in imaging through scattering media. Further, we demonstrate that this method makes it possible to simultaneously obtain multiple images and view differences in excitation parameters in a single scan of the specimen.

Download full-text


Available from: Jeff A Squier, Oct 17, 2014
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Surgical procedures as a prelude to optical imaging are a rate-limiting step in experimental neuroscience. Towards automation of these procedures, we describe the use of nonlinear optical techniques to create a thinned skull window for transcranial imaging. Metrology by second harmonic generation was used to map the surfaces of the skull and define a cutting path. Plasma-mediated laser ablation was utilized to cut bone. Mice prepared with these techniques were used to image subsurface cortical vasculature and blood flow. The viability of the brain tissue was confirmed via histological analysis and supports the utility of solely optical techniques for osteotomy and potentially other surgical procedures.
    Optics Express 10/2013; 21(20):23160-23168. DOI:10.1364/OE.21.023160 · 3.53 Impact Factor