label-free-non-linear-optics-for-study-tubulin-dependent-defects

Label-Free Non-Linear Optics for the Study of Tubulin-Dependent Defects in Central Myelin

The satisfactory visualization of cytoskeletal components in the brain is challenging. The ubiquitous distribution of the networks of microtubules, ...

Centro de Investigaciones en Óptica A.C. (CIO)

The use of electronic devices to measure Relative Humidity (RH) is widespread. However, under certain circumstances, for example when explosive gases are present, a spark-free method should be used. Here we suggest the use of stimuli-responsive materials, like gelatin and interpenetrated polymers, to detect RH with an optical method. These materials are hydrophilic. When water vapor is absorbed by the films the molecules attach to the films molecular network. The result is that the film thickness increases and their refractive index changes. To detect the change of these two parameters an optical method based on diffraction gratings is employed. Surface diffraction gratings are recorded on the films. Then gratings are placed in an optical configuration that is immersed in a climatic chamber. A light beam is sent to the grating where it is diffracted. Several light orders appear. Due to the absorption of water molecules the films swell and grating surface modulation changes. This implies that the diffracted orders intensity changes. A calibrating plot relating intensity as a function of RH is obtained.

Stimuli-Responsive Systems in Optical Humidity-Detection Devices.

Hypomyelination with atrophy of the basal ganglia and cerebellum is a recently described tubulinopathy caused by a mutation in the tubulin beta 4a isoform, expressed in oligodendrocytes. The taiep rat is the only spontaneous tubulin beta 4a mutant available for the study of this pathology. We aimed to identify the effects of the tubulin mutation on freshly collected, unstained samples of the central white matter of taiep rats using second harmonic generation microscopy. Cytoskeletal differences between the central white matter of taiep rats and control animals were found. Nonlinear emissions from the processes and somata of oligodendrocytes in tubulin beta 4a mutant rats were consistently detected, in the shape of elongated structures and cell-like bodies, which were never detected in the controls. This signal represents the second harmonic trademark of the disease. The tissue was also fluorescently labeled and analyzed to corroborate the origin of the nonlinear signal. Besides enabling the description of structural and molecular aspects of H-ABC, our data open the door to the diagnostic use of nonlinear optics in the study of neurodegenerative diseases, with the additional advantage of a label-free approach that preserves tissue morphology and vitality.

Valeria Piazza

Centro de Investigaciones en Óptica A.C. (CIO)

Label-Free Non-Linear Optics for the Study of Tubulin-Dependent Defects in Central Myelin