Journal of Physical Chemistry & Biophysics
Open Access

Multispectral topological laser speckle analyzers of proliferation and differentiation activity during morphogenesis based on tunable laser diodes and spectrometric fingerprinting of the cell cycle stages

International Conference and Trade fair on Laser Technology

July 20-22, 2015 Orlando, Florida, USA

Oganessian V.A.1, Notchenko A.V. 2, Gradov O.V.3

Posters-Accepted Abstracts: J Phys Chem Biophys

Abstract :

An automated system for morpho-topological determination of the cell division phases and structural differentiation of tissues during morphogenesis was implemented on the basis of topological properties of the cellular cultures, analyzed within the theory of sets. A simple robotic hardware and software system based on a Zeiss microscope with a modified stage and a multi-axis robotic manipulator allows to control the laser beam position, to carry out the angular irradiation of the samples using tunable laser diode sources. In the case of selective fluorescent staining which reveals the intracellular agents responsible for the histogenesis and morphogenesis regulation the use of tunable lasers with different spectral parameters allows to distinguish between the various cell types, since the topological tissue speckle patterns of different subpopulations corresponding to different wavelengths are rather different. We have currently developed a special software for the analysis and recognition of the patterns obtained from the setup described above, as well as the GUI for the precision positioning control of the stage relative to the tunable laser diode sources. Using spectrozonal overlap of the contours of the cellular speckles and the fluorescent patterns one can reveal the coherence of the functional cellular assemblies and subpopulations in tissues, cultures or an experimental matrix. This allows to obtain spectrally verifiable and derived from the local TDLS data on the metabolic relationships in a tissue or a cell culture, plasticity and adaptability of the cells (according to the gradient of the spectrochemically selective staining between different cellular points). The early prototypes of the above system without tunable diode lasers and spectral subsystems were described in while this report describes for the very first time its spectral modification based on tunable diode lasers.