Differential Multiphoton Laser Scanning Microscopy

• Published in: IEEE journal of selected topics in quantum electronics Vol. 18; no. 1; pp. 14 - 28
• Main Authors: Field, J. J., Sheetz, K. E., Chandler, E. V., Hoover, E. E., Young, M. D., Shi-you Ding, Sylvester, A. W., Kleinfeld, D., Squier, J. A.
• Format: Journal Article
• Language: English
• Published: United States IEEE 01-01-2012; IEEE Lasers and Electro-optics Society
• Subjects: Fluorescence microscopy; Laser beams; Laser excitation; Microscopy; nonlinear microscopy; Nonlinear optics; OTHER INSTRUMENTATION; Photonics; Scattering; second-harmonic generation (SHG); two-photon microscopy; ultrafast optics; ultrafast optics; nonlinear optics; fluorescence microscopy; Multiphoton microscopy; nonlinear microscopy
• ISSN: 1077-260X; 1558-4542
• DOI: 10.1109/JSTQE.2010.2077622

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.