Digital Holographic Microscopy for Characterization of Fabiana Imbricata Ruiz & Pav. Cell Suspension Cultures

P.S. Halkoglua, S.D. Yanchevaa, A.I. Pavlovb, E.M. Mihaylovaa aAgricultural University Plovdiv, Plovdiv, Bulgaria bUniversity of Food Technology, Plovdiv, Bulgaria

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An application of digital holographic microscopy (DHM) for measuring the size of cell clusters in cell suspension cultures in vitro is reported for the first time. Undifferentiated plant cell cultures are widely used for fundamental and applied research, for the elucidation of biosynthetic pathways, production of secondary metabolites and as screening tools for biotechnology in pharmacy, food technology, and agriculture. Digital in-line holographic microscopy has been applied to visualize three different cell suspensions of Fabiana imbricata Ruiz & Pav., named A, D, and MSD. Digital reconstruction of the recorded interference patterns was performed using an appropriate software. The reconstructed intensities represent the cell clusters in the suspensions under observation. Cell suspension cultures of Fabiana imbricata Ruiz & Pav. consists of cell aggregates dispersed and growing in shaking liquid media. Small cell aggregates with dimensions between 120 and 180 μm have been observed in all suspensions. The measurement of growth parameters in different cultures introduces diverse problems that must be addressed by using a specific methodology for each type of callus and cell suspension cultures. The attractive features of digital holographic microscopy are noncontact, nondestructive, marker-free in vivo imaging and quantifying biological cells and tissues. It is an advantageous technique for application in biological research and the agricultural sciences. It is shown that DIHM is a new microscopic technique easy to apply to study the size and the structure of cell aggregates in cell suspension cultures in vitro.

DOI:10.12693/APhysPolA.135.1132 topics: Fabiana imbricata, digital holographic microscopy, plant cell cultures