The use of a spaceflight-compatible device to perform WBC surface marker staining and whole-blood mitogenic activation for cytokine detection by flow cytometry.
Significant changes have recently been described regarding circulating peripheral immune cells immediately following spaceflight. Existing methods for immunophenotype staining of peripheral blood in terrestrial labs do not meet the constraints for flight on the Space Shuttle. We have recently described the development and use of the Whole Blood Staining Device (WBSD), a simple device for staining flow cytometry specimens during spaceflight. When preparing samples with the WBSD, all liquids are safely contained as the cells are moved through staining, lysis and fixation steps. Here we briefly review the use of the WBSD, and then describe another versatile adaptation, a modification to perform intracellular staining of cytokines for detection by flow cytometry. Alterations in cytokine production have been reported both in ground-based simulated microgravity culture and in astronaut samples returning from spaceflight. Data regarding microgravity effects on cytokine production for specific subpopulations of cells is lacking. Flow cytometric cytokine analysis offers the unique ability to perform simultaneous surface marker analysis and positively identity cytokine producing subsets of cells. The utilization of the WBSD provides the ability to perform rapid and routine mitogenic activation during spaceflight coupled with the ability to perform simultaneous surface marker analysis. The only external requirements for this procedure are an in-flight 37-degree incubator and the capacity for 4-degree storage.
Crucian
B E
BE
Wyle Laboratories, Houston, TX, USA.
Sams
C F
CF
D000954:Antigens, Surface / Q000032:analysis / Q000097:blood / Q000276:immunology; D016207:Cytokines / Q000032:analysis / Q000097:blood* / Q000276:immunology; D004867:Equipment Design; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000295:instrumentation*; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000276:immunology*; D013026:Space Flight / Q000295:instrumentation*; D013194:Staining and Labeling / Q000295:instrumentation*
D000954:Antigens, Surface; D016207:Cytokines