Microsampling water and blood

鱼类和两栖类胚胎在各种水生环境中发育，但个体胚胎旁边的微环境可能与自由流动的水有很大不同。 氧气将被消耗，并且诸如二氧化碳（CO2）、氢离子（H+）和氨（NH+4）的废物将被添加到与胚胎相邻的未搅拌的水层，也称为边界水层。 我要求设备（微电极，米）采样虹鳟鱼（Oncorhynchus mykiss）和蝾螈（Ambystoma maculosum）胚胎的水边界层，以确定O2，CO2，NH 4+和H+（pH值）浓度，以提供胚胎拥挤，水流和共生关系如何改变脊椎动物发育的微环境的关键信息。 为了探索水微环境的变化对生理系统的影响，我要求PeeWee 50氧分析仪测量从鱼胚胎中提取的微量血液样品的血红蛋白（Hb）对氧的亲和力。 Hb-O2亲和力提供了Hb蛋白如何响应于发育和环境影响而变化的标准量度。 总之，本申请中要求的设备将帮助我们填补我们在胚胎附近直接物理化学微环境变化如何影响其生理，生长和发育方面的知识空白。

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.