Microsampling water and blood

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Fish and amphibian embryos develop in a variety of aquatic environments but the microenvironment next to the individual embryo may be substantially different from the free-stream water.  Oxygen will be consumed and waste products such as carbon dioxide (CO2), hydrogen ions (H+) and ammonia (NH+4) will be added to the layer of unstirred water adjacent to the embryo, also termed the boundary water layer.  I request equipment (microelectrodes, meters) to sample the water boundary layer of rainbow trout (Oncorhynchus mykiss) and salamander (Ambystoma maculosum) embryos to determine O2, CO2, NH4+ and H+ (pH) concentrations to provide critical information on how embryo crowding, water flow, and symbiotic relationships can alter the microenvironment of developing vertebrates.  To explore the impact of changes in the water microenvironment on physiological systems, I request a PeeWee50 Oxygen analyzer to measure the affinity of blood hemoglobin (Hb) for oxygen on minute blood samples taken from fish embryos.  The Hb-O2 affinity provides a standard measure of how Hb proteins change in response to developmental and environmental influences.  Taken together, the equipment requested in this application will help us to fill in significant gaps in our knowledge of how changes in the immediate physico-chemical microenvironment near embryos impacts their physiology, growth and development.