1 March 2009 Extended-working-distance multiphoton micromanipulation microscope for deep-penetration imaging in live mice and tissue
Author Affiliations +
Abstract
We constructed a multiphoton (2-P) microscope with space to mount and operate microphysiology hardware, and still acquire high quality 2-P images of tumor cells deep within tissues of live mice. We reconfigured for nondescanned 2-P imaging, a dedicated electrophysiology microscope, the Nikon FN1. This microscope is compact, with retractable objectives, allowing more stage space. The instrument is fitted with long-working-distance objectives (2.5- to 3.5-mm WD) with a narrow bore, high NA, and efficient UV and IR light transmission. The system is driven by a powerful 3.5-W peak power pulsed Ti-sapphire laser with a broad tuning range. This 2-P system images a fluorescent standard to a depth of 750 to 800 μm, acquires images of murine pancreatic tumors in vivo, and also images fluorescently labeled T-cells inside live, externalized mouse lymph nodes. Effective imaging depths range between 100 and 500 μm. This compares favorably with the 100- to 300 μm micron depth attained by many 2-P systems, especially descanned 2-P instruments, and 40 μm-deep imaging with confocal microscopes. The greater depth penetration is attributable to the use of high-NA long-working-distance water-dipping lenses incorporated into a nondescanned instrument with carefully configured laser beam introduction and image-acquisition optics. Thus the new system not only has improved imaging capabilities, but allows micromanipulation and maintenance of tissues and organs.
© (2009) Society of Photo-Optical Instrumentation Engineers (SPIE)
Milan Makale, Michele McElroy, Peter O'Brien, Robert M. Hoffman, Sharon Guo, Michael Bouvet, Leo Barnes, Elizabeth Ingulli, David Cheresh, "Extended-working-distance multiphoton micromanipulation microscope for deep-penetration imaging in live mice and tissue," Journal of Biomedical Optics 14(2), 024032 (1 March 2009). https://doi.org/10.1117/1.3103783 . Submission:
JOURNAL ARTICLE
10 PAGES


SHARE
Back to Top