Microfluidic Device for Long-Term Zebrafish Heart Culture and Imaging
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Solution Overview
Problem
Current experimental tools are inadequate for investigating adult zebrafish heart regeneration, as they fail to maintain organ structure and function for extended periods, limiting the ability to study cell-cell interactions and organ-level mechanisms, and are not compatible with continuous live imaging.
Innovation Solution
A microfluidic chip system that maintains juvenile or adult zebrafish hearts in a stable position, provides continuous media flow, and is compatible with conventional imaging systems, allowing for long-term culture and live imaging of organ regeneration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If explanted hearts are cultured in Petri dishes or multi-well plates, then the hearts can be cultured ex vivo, but the native-like morphology and function rapidly decline within three days
Solution Approach 1:
The culture system is segmented into multiple functional zones: a reservoir for media storage, channels for controlled media delivery, and imaging chambers for observation. This segmentation allows different regions to perform specialized functions, maintaining heart viability longer through optimized media distribution and waste removal.
Solution Approach 2:
The system uses hydraulic principles to deliver culture media through channels to the explanted heart, enabling controlled perfusion that mimics physiological conditions. This fluid delivery system maintains morphological and functional integrity by providing continuous nutrient supply and removing metabolic waste, extending culture lifetime beyond the three-day limit of static cultures.
2Reliability
If hearts undergo gentle agitation in culture dishes, then morphological and functional declines are partially alleviated, but the hearts are not confined to a specific location or orientation making continuous live imaging incompatible
Solution Approach 1:
The system transitions from two-dimensional Petri dish culture to a three-dimensional microfluidic environment with controlled fluid flow. This dimensional change allows the heart to remain stationary in a specific orientation within the imaging chamber while media is delivered through channels, simultaneously achieving both improved reliability and imaging compatibility.
Solution Approach 2:
The microfluidic channels act as intermediaries between the media reservoir and the explanted heart, delivering nutrients and removing waste without requiring physical agitation of the heart itself. This intermediary system maintains morphological integrity while keeping the heart stationary for imaging.
3Ease of operation
If microfabricated fluidic devices are used to culture zebrafish embryos and larvae, then continuous live imaging is facilitated by minimizing arbitrary motion, but the devices are not optimized for long-term culture of explanted adult hearts
Solution Approach 1:
The device is designed with universal features that can accommodate different types of biological samples including both zebrafish embryos and explanted adult hearts. The imaging chamber geometry, media channels, and confinement structures are optimized to work with various organ sizes and shapes, extending the application of microfluidic imaging technology to long-term adult heart culture.
Solution Approach 2:
The device parameters such as chamber dimensions, channel flow rates, and media composition are optimized specifically for adult heart culture requirements. By adjusting these parameters from embryo-culture settings to heart-culture settings, the system extends culture lifetime while maintaining imaging compatibility.
Data Source
AI summary
Zebrafish are a powerful model for investigating cardiac repair due to their unique regenerative abilities, scalability, and compatibility with many genetic tools. However, characterizing the regeneration process in live adult zebrafish hearts has proved challenging because adult fish are opaque and explanted hearts in conventional culture conditions experience rapid declines in morphology and physiology. To overcome these limitations, we fabricated a fluidic device for culturing explanted adult zebrafish hearts with constant media perfusion that is also compatible with live imaging. Unlike hearts cultured in dishes for one week, the morphology and calcium activity of hearts cultured in the device for one week were largely similar to freshly explanted hearts. We also cultured injured hearts in the device and used live imaging techniques to continuously record the revascularization process over several days, demonstrating how our device enables unprecedented visual access to the multi-day process of adult zebrafish heart regeneration.