Phan, M. S., Matho, K., Abdeladim, L., Livet, J., Beaurepaire, E., Chessel, A. (April 2019) Automated quantification with sub-micrometer scale precision in volumetric multicolor multiphoton microscopy images. Institute of Electrical and Electronics Engineers, 521-525; art no 8759200. ISBN 19457928 (ISSN); 9781538636411 (ISBN)
Abstract
Recent multicolor multiphoton microscopy schemes are opening the way to high quality fluorescence color imaging at near-micrometric resolution over virtually unlimited volumes of brain tissue. However, unavoidable artifacts related to depth, chromatic aberration or anisotropic resolution affect the resulting images and can bias measurements and quantifications. Here, we prove the possibility of performing precise automated quantitative measurements on such tridimensional multicolor images, taking as a model fluorescently labeled axons belonging to an auditory tract in the mouse brainstem. We present an analysis pipeline to compute the diameter of these axons based on the calculation of distance in HSV color space, its binarization using a random walker algorithm and skeleton extraction by distance transform. We measure a biologically meaningful difference of about 500 nm before and after the axons enter their target nucleus where they form synaptic connections, proving the robustness of our pipeline with respect to the aforementioned limitations. This demonstrates the ability of sub-micrometric measurements across tissue depths of 0.5 mm with multicolor multiphoton microscopy. © 2019 IEEE.
| Item Type: | Book |
|---|---|
| Additional Information: | Conference |
| Subjects: | organism description > animal > mammal > rodent > mouse Investigative techniques and equipment > microscopy > flourescence microscopy > two-photon excitation microscopy |
| CSHL Authors: | |
| Depositing User: | Matthew Dunn |
| Date: | April 2019 |
| Date Deposited: | 08 Nov 2019 20:15 |
| Last Modified: | 08 Nov 2019 20:15 |
| PMCID: | 2019 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/38658 |
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