Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material
Oxygen equilibrium curves have been widely used to understand oxygen transport in numerous organisms. A major challenge has been to monitor oxygen binding characteristics and concomitant pH changes as they occur in vivo, in limited sample volumes. Here we report a technique allowing highly resolved...
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ftpangaea:oai:pangaea.de:doi:10.1594/PANGAEA.831205 2024-09-15T17:45:29+00:00 Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material Oellermann, Michael Pörtner, Hans-Otto Mark, Felix Christopher 2014 application/zip, 4.1 MBytes https://doi.pangaea.de/10.1594/PANGAEA.831205 https://doi.org/10.1594/PANGAEA.831205 en eng PANGAEA https://doi.pangaea.de/10.1594/PANGAEA.831205 https://doi.org/10.1594/PANGAEA.831205 CC-BY-3.0: Creative Commons Attribution 3.0 Unported Access constraints: unrestricted info:eu-repo/semantics/openAccess Supplement to: Oellermann, Michael; Pörtner, Hans-Otto; Mark, Felix Christopher (2014): Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in blood microvolumes. Journal of Experimental Biology, 217(9), 1430-1436, https://doi.org/10.1242/jeb.092726 Priority Programme 1158 Antarctic Research with Comparable Investigations in Arctic Sea Ice Areas SPP1158 dataset 2014 ftpangaea https://doi.org/10.1594/PANGAEA.83120510.1242/jeb.092726 2024-07-24T02:31:32Z Oxygen equilibrium curves have been widely used to understand oxygen transport in numerous organisms. A major challenge has been to monitor oxygen binding characteristics and concomitant pH changes as they occur in vivo, in limited sample volumes. Here we report a technique allowing highly resolved and simultaneous monitoring of pH and blood pigment saturation in minute blood volumes. We equipped a gas diffusion chamber with a broad range fibre optic spectrophotometer and a micro-pH optode and recorded changes of pigment oxygenation along PO2 and pH gradients to test the setup. Oxygen binding parameters derived from measurements in only 15 µl of haemolymph from the cephalopod Octopus vulgaris showed low instrumental error (0.93%) and good agreement with published data. Broad range spectra, each resolving 2048 data points, provided detailed insight into the complex absorbance characteristics of diverse blood types. After consideration of photobleaching and intrinsic fluorescence, pH optodes yielded accurate recordings and resolved a sigmoidal shift of 0.03 pH units in response to changing PO2 from 0-21 kPa. Highly resolved continuous recordings along pH gradients conformed to stepwise measurements at low rates of pH changes. In this study we showed that a diffusion chamber upgraded with a broad range spectrophotometer and an optical pH sensor accurately characterizes oxygen binding with minimal sample consumption and manipulation. We conclude that the modified diffusion chamber is highly suitable for experimental biologists who demand high flexibility, detailed insight into oxygen binding as well as experimental and biological accuracy combined in a single set up. Dataset Antarc* Antarctic Sea ice PANGAEA - Data Publisher for Earth & Environmental Science |
institution |
Open Polar |
collection |
PANGAEA - Data Publisher for Earth & Environmental Science |
op_collection_id |
ftpangaea |
language |
English |
topic |
Priority Programme 1158 Antarctic Research with Comparable Investigations in Arctic Sea Ice Areas SPP1158 |
spellingShingle |
Priority Programme 1158 Antarctic Research with Comparable Investigations in Arctic Sea Ice Areas SPP1158 Oellermann, Michael Pörtner, Hans-Otto Mark, Felix Christopher Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
topic_facet |
Priority Programme 1158 Antarctic Research with Comparable Investigations in Arctic Sea Ice Areas SPP1158 |
description |
Oxygen equilibrium curves have been widely used to understand oxygen transport in numerous organisms. A major challenge has been to monitor oxygen binding characteristics and concomitant pH changes as they occur in vivo, in limited sample volumes. Here we report a technique allowing highly resolved and simultaneous monitoring of pH and blood pigment saturation in minute blood volumes. We equipped a gas diffusion chamber with a broad range fibre optic spectrophotometer and a micro-pH optode and recorded changes of pigment oxygenation along PO2 and pH gradients to test the setup. Oxygen binding parameters derived from measurements in only 15 µl of haemolymph from the cephalopod Octopus vulgaris showed low instrumental error (0.93%) and good agreement with published data. Broad range spectra, each resolving 2048 data points, provided detailed insight into the complex absorbance characteristics of diverse blood types. After consideration of photobleaching and intrinsic fluorescence, pH optodes yielded accurate recordings and resolved a sigmoidal shift of 0.03 pH units in response to changing PO2 from 0-21 kPa. Highly resolved continuous recordings along pH gradients conformed to stepwise measurements at low rates of pH changes. In this study we showed that a diffusion chamber upgraded with a broad range spectrophotometer and an optical pH sensor accurately characterizes oxygen binding with minimal sample consumption and manipulation. We conclude that the modified diffusion chamber is highly suitable for experimental biologists who demand high flexibility, detailed insight into oxygen binding as well as experimental and biological accuracy combined in a single set up. |
format |
Dataset |
author |
Oellermann, Michael Pörtner, Hans-Otto Mark, Felix Christopher |
author_facet |
Oellermann, Michael Pörtner, Hans-Otto Mark, Felix Christopher |
author_sort |
Oellermann, Michael |
title |
Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
title_short |
Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
title_full |
Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
title_fullStr |
Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
title_full_unstemmed |
Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
title_sort |
simultaneous high-resolution ph and spectrophotometric recordings of oxygen binding in native blood microvolumes, link to supplementary material |
publisher |
PANGAEA |
publishDate |
2014 |
url |
https://doi.pangaea.de/10.1594/PANGAEA.831205 https://doi.org/10.1594/PANGAEA.831205 |
genre |
Antarc* Antarctic Sea ice |
genre_facet |
Antarc* Antarctic Sea ice |
op_source |
Supplement to: Oellermann, Michael; Pörtner, Hans-Otto; Mark, Felix Christopher (2014): Simultaneous high-resolution pH and spectrophotometric recordings of oxygen binding in blood microvolumes. Journal of Experimental Biology, 217(9), 1430-1436, https://doi.org/10.1242/jeb.092726 |
op_relation |
https://doi.pangaea.de/10.1594/PANGAEA.831205 https://doi.org/10.1594/PANGAEA.831205 |
op_rights |
CC-BY-3.0: Creative Commons Attribution 3.0 Unported Access constraints: unrestricted info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1594/PANGAEA.83120510.1242/jeb.092726 |
_version_ |
1810493333283274752 |