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Published online on May 23, 2003, 10.1073/pnas.0731953100
PNAS | June 10, 2003 | vol. 100 | no. 12 | 7081-7086


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Biophysics
Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy

Daniel A. Dombeck *, Karl A. Kasischke *, Harshad D. Vishwasrao *, Martin Ingelsson {dagger}, Bradley T. Hyman {dagger}, and Watt W. Webb * {ddagger}

*School of Applied and Engineering Physics, Cornell University, Ithaca, NY 14853; and {dagger}Harvard Medical School, Alzheimer's Disease Research Center, Massachusetts General Hospital, Charlestown, MA 02129

Contributed by Watt W. Webb, April 4, 2003

Microtubule (MT) ensemble polarity is a diagnostic determinant of the structure and function of neuronal processes. Here, polarized MT structures are selectively imaged with second-harmonic generation (SHG) microscopy in native brain tissue. This SHG is found to colocalize with axons in both brain slices and cultured neurons. Because SHG arises only from noninversion symmetric structures, the uniform polarity of axonal MTs leads to the observed signal, whereas the mixed polarity in dendrites leads to destructive interference. SHG imaging provides a tool to investigate the kinetics and function of MT ensemble polarity in dynamic native brain tissue structures and other subcellular motility structures based on polarized MTs.

Abbreviations: MT, microtubule; SH, second harmonic; SHG, SH generation; TPF, two-photon fluorescence; NA, numerical aperture; PMT, photo-multiplier tube; DG, dentate gyrus; <I>, average excitation intensity; w, two-photon radial beam waist; MAP, MT-associated protein.

{ddagger} To whom correspondence should be addressed. E-mail: www2{at}cornell.edu.


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