TY - JOUR
T1 - Module to support real-time microscopic imaging of living organisms on ground-based microgravity analogs
AU - Neelam, Srujana
AU - Lee, Audrey
AU - Lane, Michael A.
AU - Udave, Ceasar
AU - Levine, Howard G.
AU - Zhang, Ye
N1 - Funding Information:
organisms to stimuli. Microgravity and hypergravity are two mechanical stimuli that hu- Supplementary Materials: The following are available online at https://www.mdpi.com/article/10 mans and plants are exposed to on space missions. Several studies have been performed .3390/app11073122/s1, Video S1. using live-cell imaging capability on the ISS and sounding rocket true microgravity platforms to understand the response of human cells to microgravity [30,31]. In order to provide a more cost-effective and easily accessible platform to researchers, we developed a live-imaging module to capture the dynamics of cells in SMG. A rotating machine pro- preparation, S.N. and A.L.; writing—review and editing, Y.Z., H.G.L. and S.N.; visualization, Y.Z., vides a challenging environment for any optical-mechanical setup because loading levels S.N. and A.L.; supervision, S.N. and Y.Z.; project administration, S.N. and Y.Z.; funding acquisition continuously change. Accelerations and rotation direction changes dramatize the condi- Y.Z. and H.G.L. All authors have read and agreed to the published version of the manuscript. tion by introducing mechanical shocks. The module described herein successfully addresses these challenges and permits the live imaging of cells over prolonged intervals. Srujana Neelam was supported by the NASA Post-doctoral (NPP) program (NNH15C048B) adminis- Supplementary Materials: The following are available online at www.mdpi.com/xxx/s1, Video S1. writing, and her participation in editing and revision was supported by the NASA Space Biology progAraumth,oGrrCanotn#tr8i0bNuStiSoCn2s0:KC0o4n2c3e.pAtuuadlriezyatLioene,aSn.Nd .Caenadsa Yr.UZd.; amveetwhoedreolsougpyp,oSr.tNed., bAy.Lt.h, eCN.UA.;S sAofStwpaacree, A.L., C.U. and M.A.L.; validation, S.N., A.L. and M.A.L.; formal analysis, S.N. and A.L.; investigation, S.N., A.L. and C.U.; resources, Y.Z. and H.G.L.; data curation, Y.Z. and H.G.L.; writing—original draft preparation, S.N. and A.L.; writing—review and editing, Y.Z., H.G.L. and S.N.; visualization, Y.Z., S.N. and A.L.; supervision, S.N. and Y.Z.; project administration, S.N. and Y.Z.; funding acquisition Y.Z. and H.G.L. All authors have read and agreed to the published version of the manuscript.
Funding Information:
The MSSF and this development were funded by the NASA Space Biology Program. Srujana Neelam was supported by the NASA Post-doctoral (NPP) program (NNH15C048B) administered by Universities Space Research Association (USRA) for the whole development and manuscript writing, and her participation in editing and revision was supported by the NASA Space Biology program, Grant #80NSSC20K0423 Audrey Lee and Ceasar Udave were supported by the NASA Space Biology Program through the NASA internship program. This development was submitted in the NASA New Technology Reporting System as KSC-14221 on 28 September 2018, entitled ?Modules to Support Live Microscopic Imaging and Samples on Ground-Based Microgravity Simulator Devices?.
Publisher Copyright:
© 2021 by the authors.
PY - 2021/4/1
Y1 - 2021/4/1
N2 - 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.
AB - 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.
KW - Cell culture
KW - Live cell imaging
KW - Microgravity simulation
KW - Random positioning machine
UR - https://www.scopus.com/pages/publications/85104057487
U2 - 10.3390/app11073122
DO - 10.3390/app11073122
M3 - Article
AN - SCOPUS:85104057487
SN - 2076-3417
VL - 11
JO - Applied Sciences (Switzerland)
JF - Applied Sciences (Switzerland)
IS - 7
M1 - 3122
ER -