The FixBox: Hardware to Provide on-Orbit Fixation Capabilities to the EMCS on the ISS

Plant biology is an important area for the future of space exploration, but biological spaceflight experiments have been always constrained by the hardware capabilities. The European Modular Cultivation System (EMCS) unit was an incubator for small organisms, such as Arabidopsis thaliana, built by the European Space Agency (ESA) and was decommissioned in 2018. Here, we describe the FixBox system as add-on hardware to provide fixation capabilities to the plant growth cassettes, which, initially, were not designed to be used for that purpose. Tests were performed to ensure the successful use of this device in the EMCS facility. We also evaluate the required adaptations to the hardware, e.g., to guarantee that the reduced fluid motion in microgravity does not cause any bubbles that could impair the quality of fixation. Arabidopsis thaliana seedlings grown during spaceflight were fixed in the FixBox either in glutaraldehyde or formaldehyde. Electron microscopical images and confocal microscopy immunofluorescent localizations showed an excellent preservation of both cell ultrastructure and antigen conformation. Thus, it is possible to modify existing hardware to comply with the scientific requirements to augment the existing capabilities on board the ISS. In addition, it is also possible to reuse culture chambers from predesigned experimental containers into new modular subunits as FixBox. Similarly, we can design new hardware compatible with a novel cultivation chamber on board, such as is available in BIOLAB, to be used later with FixBox. Lessons learned for future space plant biology researchers include how to manage the number of hardware requirements and constraints on how to preserve the biological samples.

[1]  Gilbert Gasset,et al.  Plant cell proliferation and growth are altered by microgravity conditions in spaceflight. , 2010, Journal of plant physiology.

[2]  V. Pereda-Loth,et al.  Both gravistimulation onset and removal trigger an increase of cytoplasmic free calcium in statocytes of roots grown in microgravity , 2018, Scientific Reports.

[3]  Richard E. Edelmann,et al.  Changes in operational procedures to improve spaceflight experiments in plant biology in the European Modular Cultivation System , 2014 .

[4]  Richard E. Edelmann,et al.  Ground-based studies of tropisms in hardware developed for the European Modular Cultivation System (EMCS) , 2005 .

[5]  Susan Hutchison,et al.  Seven Years of Permanent Running of MELFI-1 on Board the ISS and Utilisation of the Three MELFI Units Refrigeration Pool , 2013 .

[6]  Eugénie Carnero-Diaz,et al.  Gravisensitivity and automorphogenesis of lentil seedling roots grown on board the International Space Station. , 2008, Physiologia plantarum.

[7]  Richard E. Edelmann,et al.  Operations of a spaceflight experiment to investigate plant tropisms , 2009 .

[8]  Raymond M. Wheeler,et al.  Microbiological and Nutritional Analysis of Lettuce Crops Grown on the International Space Station , 2020, Frontiers in Plant Science.

[9]  Howard G. Levine,et al.  Growth Chambers on the International Space Station for Large Plants , 2016 .

[10]  David W. Reed,et al.  The performance of KSC Fixation Tubes with RNALater for orbital experiments: A case study in ISS operations for molecular biology , 2011 .

[11]  R. Herranz,et al.  Exploration of plant growth and development using the European Modular Cultivation System facility on the International Space Station. , 2014, Plant biology.

[12]  Francisco Javier Medina,et al.  Nucleolar structure and proliferation activity of Arabidopsis root cells from seedlings germinated on the International Space Station , 2005 .

[13]  S. Wyatt,et al.  Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana , 2017, PloS one.

[14]  George C. Lisensky,et al.  PREPARATION AND PROPERTIES OF AN AQUEOUS FERROFLUID , 1999 .

[15]  Richard E. Edelmann,et al.  Transcriptome analyses of Arabidopsis thaliana seedlings grown in space: implications for gravity-responsive genes , 2013, Planta.

[16]  Manuel Echeverria,et al.  Characterization of AtNUC-L1 reveals a central role of nucleolin in nucleolus organization and silencing of AtNUC-L2 gene in Arabidopsis. , 2006, Molecular biology of the cell.

[17]  M. A. Hayat,et al.  13 – Fixation for Scanning Electron Microscopy , 1981 .

[18]  G Richoilley,et al.  The CYTOS biological experiments carried out on the Soviet orbital station Salyut 6. , 1981, Aviation, space, and environmental medicine.

[19]  Tom Beeckman,et al.  A novel sensor to map auxin response and distribution at high spatio-temporal resolution , 2012, Nature.

[20]  E Brinckmann,et al.  Experiments with small animals in BIOLAB and EMCS on the International Space Station. , 2002, Advances in space research : the official journal of the Committee on Space Research.

[21]  J. Stockert,et al.  Observations on nucleolar staining with osmium tetroxide , 1974, Experientia.

[22]  E Brinckmann.  Spaceflight opportunities on the ISS for plant research--the ESA perspective. , 1999, Advances in space research : the official journal of the Committee on Space Research.

[23]  E. Brinckmann,et al.  ESA hardware for plant research on the International Space Station , 2005 .

[24]  J. Kiss,et al.  Gravitropism and development of wild-type and starch-deficient mutants of Arabidopsis during spaceflight. , 1998, Physiologia plantarum.

[25]  Richard E. Edelmann,et al.  Biocompatibility studies in preparation for a spaceflight experiment on plant tropisms (TROPI) , 2007 .

[26]  M. A. Hayat,et al.  Fixation for Electron Microscopy , 1982 .

[27]  Joshua P Vandenbrink,et al.  Preparation of a Spaceflight Experiment to Study Tropisms in Arabidopsis Seedlings on the International Space Station. , 2019, Methods in molecular biology.