Construction and Establishment of a New Environmental Chamber to Study Real-Time Cardiac Development

Abstract

Heart development, especially the critical phase of cardiac looping, is a complex and intricate process that has not yet been visualized "live" over long periods of time. We have constructed and established a new environmental incubator chamber that provides stable conditions for embryonic development with regard to temperature, humidity, and oxygen levels. We have integrated a video microscope in the chamber to visualize the developing heart in real time and present the first "live" recordings of a chick embryo in shell-less culture acquired over a period of 2 days. The time-lapse images we show depict a significant time window that covers the most critical and typical morphogenetic events during normal cardiac looping. Our system is of interest to researchers in the field of embryogenesis, as it can be adapted to a variety of animal models for organogenesis studies including heart and limb development.

Automated Multichamber Time-lapse Videography for Long-term In Vivo Observation of Migrating Cells

“…'Homemade' environmental chambers have been documented since the early 1950s (1-7) with various chamber types, many of which closely resemble modern designs (8)(9)(10)(11). More recently, environmental chambers have been combined with time-lapse videography to study living cells (12)(13)(14)(15)(16). The basic demands on such a chamber include maintenance of temperature, humidity, and atmosphere (generally 5% CO 2 in air), as well as precise and reproducible chamber positioning in order to simultaneously observe selected areas.…”

Investigating developmental cardiovascular biomechanics and the origins of congenital heart defects

“…The imaging environment varied among the different studies from room air (Bain et al, 2007 ) to full temperature and humidity control (Henning et al, 2011 ). Long-term, time-lapse imaging of live embryos, where a single embryo is kept within an imaging platform and followed continuously for a period of hours or days, has been achieved by constructing controllable chambers to maintain physiologic environmental conditions (Orhan et al, 2007 ; Gargesha et al, 2009 ; Kulesa et al, 2010 ; Ma et al, 2010 ; Happel et al, 2011 ; Al Naieb et al, 2012 ). The chick embryo has been studied using long-term, time-lapse techniques combined with both confocal microscopy to track moving cell populations over a period of more than 26 h, in 1.5 min intervals (El-Ghali et al, 2010 ; Kulesa et al, 2010 ) and OCT to measure cardiac function over a 6 h period, in 60 min intervals (Happel et al, 2011 ).…”

In vivo imaging of the cyclic changes in cross‐sectional shape of the ventricular segment of pulsating embryonic chick hearts at stages 14 to 17: A contribution to the understanding of the ontogenesis of cardiac pumping function

“…Moreover, as in our previous study, the acquisition of the present data was still made outside of an incubator under nonphysiological environmental conditions, which were expected to distort quantitative data (see the Experimental Procedures section). To overcome the latter limitations, we are currently working on the integration of the scanning unit of our OCT-system into an environmental chamber that provides stable physiological conditions (Orhan et al, 2007).…”