Abstract
Since opportunities for spaceflight experiments are scarce, ground-based microgravity simulation devices (MSDs) offer accessible and economical alternatives for gravitational biology studies. Among the MSDs, the random positioning machine (RPM) provides simulated microgravity conditions on the ground by randomizing rotating biological samples in two axes to distribute the Earth’s gravity vector in all directions over time. Real-time microscopy and image acquisition during microgravity simulation are of particular interest to enable the study of how basic cell functions, such as division, migration, and proliferation, progress under altered gravity conditions. However, these capabilities have been difficult to implement due to the constantly moving frames of the RPM as well as mechanical noise. Therefore, we developed an image acquisition module that can be mounted on an RPM to capture live images over time while the specimen is in the simulated microgravity (SMG) environment. This module integrates a digital microscope with a magnification range of 20× to 700×, a high-speed data transmission adaptor for the wireless streaming of time-lapse images, and a backlight illuminator to view the sample under brightfield and darkfield modes. With this module, we successfully demonstrated the real-time imaging of human cells cultured on an RPM in brightfield, lasting up to 80 h, and also visualized them in green fluorescent channel. This module was successful in monitoring cell morphology and in quantifying the rate of cell division, cell migration, and wound healing in SMG. It can be easily modified to study the response of other biological specimens to SMG.
Highlights
The spaceflight environment influences physiological changes in biological molecules, cells, tissues, and organs
As space exploration progresses toward long-duration crewed missions to the Moon and Mars, it is important to understand the mechanisms by which microgravity affects physiology in order to accurately assess health risks and to develop effective countermeasures
We have successfully developed and demonstrated, for the first time, a novel module that is suitable for real-time brightfield and fluorescence microscopic imaging of living organisms undergoing microgravity simulation on a desktop random positioning machine (RPM)
Summary
The spaceflight environment influences physiological changes in biological molecules, cells, tissues, and organs. The effects of SMG on samples may be functionally comparable to that of true microgravity if the gravity vector’s direction changes faster than the sample’s response time to the change [5]. Under these conditions, living objects such as mammalian cells and plants are suitable candidates for microgravity simulation studies [6]. We have successfully developed and demonstrated, for the first time, a novel module that is suitable for real-time brightfield and fluorescence microscopic imaging of living organisms undergoing microgravity simulation on a desktop RPM. Our results highlight the potential application of real-time microscopic observation and imaging in microgravity simulation devices to study the impact of altered gravity on cellular and subcellular processes
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