TY - JOUR
T1 - Glutaraldehyde - A subtle tool in the investigation of healthy and pathologic red blood cells
AU - Abay, Asena
AU - Simionato, Greta
AU - Chachanidze, Revaz
AU - Bogdanova, Anna
AU - Hertz, Laura
AU - Bianchi, Paola
AU - van den Akker, Emile
AU - von Lindern, Marieke
AU - Leonetti, Marc
AU - Minetti, Giampaolo
AU - Wagner, Christian
AU - Kaestner, Lars
N1 - Funding Information: The research leading to these results has received funding from the European Framework “Horizon 2020” under grant agreement number 675115 (RELEVANCE) and from the Volkswagen Foundation (Az: 93839). Publisher Copyright: Copyright © 2019 Abay, Simionato, Chachanidze, Bogdanova, Hertz, Bianchi, van den Akker, von Lindern, Leonetti, Minetti, Wagner and Kaestner. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
PY - 2019
Y1 - 2019
N2 - Glutaraldehyde is a well-known substance used in biomedical research to fix cells. Since hemolytic anemias are often associated with red blood cell shape changes deviating from the biconcave disk shape, conservation of these shapes for imaging in general and 3D-imaging in particular, like confocal microscopy, scanning electron microscopy or scanning probe microscopy is a common desire. Along with the fixation comes an increase in the stiffness of the cells. In the context of red blood cells this increased rigidity is often used to mimic malaria infected red blood cells because they are also stiffer than healthy red blood cells. However, the use of glutaraldehyde is associated with numerous pitfalls: (i) while the increase in rigidity by an application of increasing concentrations of glutaraldehyde is an analog process, the fixation is a rather digital event (all or none); (ii) addition of glutaraldehyde massively changes osmolality in a concentration dependent manner and hence cell shapes can be distorted; (iii) glutaraldehyde batches differ in their properties especially in the ratio of monomers and polymers; (iv) handling pitfalls, like inducing shear artifacts of red blood cell shapes or cell density changes that needs to be considered, e.g., when working with cells in flow; (v) staining glutaraldehyde treated red blood cells need different approaches compared to living cells, for instance, because glutaraldehyde itself induces a strong fluorescence. Within this paper we provide documentation about the subtle use of glutaraldehyde on healthy and pathologic red blood cells and how to deal with or circumvent pitfalls.
AB - Glutaraldehyde is a well-known substance used in biomedical research to fix cells. Since hemolytic anemias are often associated with red blood cell shape changes deviating from the biconcave disk shape, conservation of these shapes for imaging in general and 3D-imaging in particular, like confocal microscopy, scanning electron microscopy or scanning probe microscopy is a common desire. Along with the fixation comes an increase in the stiffness of the cells. In the context of red blood cells this increased rigidity is often used to mimic malaria infected red blood cells because they are also stiffer than healthy red blood cells. However, the use of glutaraldehyde is associated with numerous pitfalls: (i) while the increase in rigidity by an application of increasing concentrations of glutaraldehyde is an analog process, the fixation is a rather digital event (all or none); (ii) addition of glutaraldehyde massively changes osmolality in a concentration dependent manner and hence cell shapes can be distorted; (iii) glutaraldehyde batches differ in their properties especially in the ratio of monomers and polymers; (iv) handling pitfalls, like inducing shear artifacts of red blood cell shapes or cell density changes that needs to be considered, e.g., when working with cells in flow; (v) staining glutaraldehyde treated red blood cells need different approaches compared to living cells, for instance, because glutaraldehyde itself induces a strong fluorescence. Within this paper we provide documentation about the subtle use of glutaraldehyde on healthy and pathologic red blood cells and how to deal with or circumvent pitfalls.
KW - Batch variation
KW - Cell shapes
KW - Erythrocytes
KW - Fixation
KW - Glutaraldehyde
KW - Hemolytic anemia
KW - Osmolality
KW - Stiffness
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85068214479&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/31139090
U2 - https://doi.org/10.3389/fphys.2019.00514
DO - https://doi.org/10.3389/fphys.2019.00514
M3 - Article
C2 - 31139090
SN - 1664-042X
VL - 10
JO - Frontiers in physiology
JF - Frontiers in physiology
IS - MAY
M1 - 514
ER -