Dynamic phase differences based on quantitative phase imaging for the objective evaluation of cell behavior.
Quantitative phase imaging (QPI) brought innovation to noninvasive observation of live cell dynamics seen as cell behavior. Unlike the Zernike phase contrast or differential interference contrast, QPI provides quantitative information about cell dry mass distribution. We used such data for objective evaluation of live cell behavioral dynamics by the advanced method of dynamic phase differences (DPDs). The DPDs method is considered a rational instrument offered by QPI. By subtracting the antecedent from the subsequent image in a time-lapse series, only the changes in mass distribution in the cell are detected. The result is either visualized as a two dimensional color-coded projection of these two states of the cell or as a time dependence of changes quantified in picograms. Then in a series of time-lapse recordings, the chain of cell mass distribution changes that would otherwise escape attention is revealed. Consequently, new salient features of live cell behavior should emerge. Construction of the DPDs method and results exhibiting the approach are presented. Advantage of the DPDs application is demonstrated on cells exposed to an osmotic challenge. For time-lapse acquisition of quantitative phase images, the recently developed coherence-controlled holographic microscope was employed.
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech RepublicbBrno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600.
Aneta
A
Krizova
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech RepublicbBrno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600.
Jana
J
Collakova
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech RepublicbBrno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600.
Zbynek
Z
Dostal
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech Republic.
Lukas
L
Kvasnica
University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093, United States.
Hana
H
Uhlirova
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech RepublicbBrno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600.
Tomas
T
Zikmund
Brno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600, Czech Republic.
Pavel
P
Vesely
Brno University of Technology, Institute of Physical Engineering, Faculty of Mechanical Engineering, Technicka 2896/2, Brno 61600, Czech RepublicbBrno University of Technology, CEITEC-Central European Institute of Technology, Technicka 3058/10, Brno 61600.
Radim
R
Chmelik
D000818:Animals; D002460:Cell Line; D048430:Cell Shape / Q000502:physiology; D003584:Cytological Techniques / Q000379:methods*; D006696:Holography / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D009997:Osmotic Pressure / Q000502:physiology; D051381:Rats