Journal: Physiological Reports

Article Title: High‐throughput determination of oxygen dissociation curves in a microplate reader—A novel, quantitative approach

doi: 10.14814/phy2.14995

Figure Lengend Snippet: Scheme of the method to determine, in parallel, ODC of small blood samples by a modified 96‐well plate and a microplate reader. (a) ODC plate. All wells are interspaced with a black polymer, and channels milled to form a meandering gas flow system when covered with a transparent seal. Wells are either empty (white) or contain thin films of whole blood (pink), internal standards (red) and two O 2 sensors (blue) as depicted. Signals of the in‐ and outflow O 2 sensors (blue line graphs) and two (A2 and A4), out of 92, exemplary ODCs are shown (top). (b) Cross‐section through wells H5‐F5 with details of the polymer cast and the gas channel system, as well as principle of measurements (fluorescence intensity in the bottom reading mode for oxygen partial pressure, P O 2 , absorption measurements for oxygen saturation, S O 2 ). (c) Microscopy of the central blood film showing a 1–2 cell layer of red blood cells over large parts. (d) Entire setup with computer, gas supply, temperature‐controlled humidifying gas reservoir, modified cover, microplate reader (front view), and a peristaltic pump (bottom right). (e) Top‐view of the modified cover allowing a flexible, smooth, and light‐proof connection between the ODC plate and gas supply/outflow. Arrows indicate movements of the plate within the microplate reader during measurements. (f) Depicts an exemplary possibility for modifying the ODC plate with four independent (colors) gas flow systems (= four‐channel ODC plate). ODC, oxygen dissociation curve

Article Snippet: After plating the RBC, the top of the plate was sealed with a transparent, adhesive microplate sealer (Greiner AG), providing unidirectional gas flow along the channels and gas tightness between the wells and the exterior (Figure ).

Techniques: Modification, Fluorescence, Microscopy