2013 ASCB Annual Meeting in New Orleans

Date: 14th -18th December 2013
Place: New Orleans, LA, USA

Dynamics of cell behavior by Coherence Controlled Holographic Microscope (CCHM)
Collakova Jana, Krizova Aneta, Dostal Zbynek, Kvasnica Lukas, Slaby Tomas, Zikmund Tomas, Kolman Pavel, Uhlirova Hana, Vesely Pavel, Chmelik Radim

CCHM is a new digital holographic microscope designed to work with a wide coherence range of the illumination (Kolman et al., Opt Express (2010); Slaby et al., Opt Express (2013)). Its non-invasive, instantaneous quantitative phase imaging is used here for visualization and measurement of cell dynamics. Moreover, the inherent coherence gating effect enables quantitative imaging even in turbid media which makes this microscope an invaluable tool for imaging cells through thick, scattering milieu. A procedure for numerical reconstruction and segmentation of cells is described here. Once cells have been segmented, a method of dynamic phase difference is applied for the evaluation of cell migration and motility. This yields a printable 2D record of time-lapsed cell motility and also a calculation of dry mass changes per unit of time. The rate of cell growth and/or progression of cell movements measured in picograms per time enables statistical evaluation of cellular activity.
We introduce the potential of CCHM for cell biology by imaging and measurement of cell growth, motility and migration on a planar surface and in 3D structures of turbid media – a collagen gel and a phospholipid emulsion. Non-labelled malignant cells are imaged through a 1 mm-thick layer of 0.2% collagen or up to a 4 mm thick layer of 0.15% phospholipid emulsion demonstrating the possibility to study their reactions.