Cell Fixation by Light-Triggered Release of Glutaraldehyde
Dr. Korwin M. Schelkle
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
These authors contributed equally to this work.
Search for more papers by this authorChristopher Schmid
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
These authors contributed equally to this work.
Search for more papers by this authorM. Sc. Klaus Yserentant
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Physikalisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 229, 69120 Heidelberg, Germany
CellNetworks, Single Molecule Spectroscopy, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorM. Sc. Markus Bender
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Irene Wacker
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Martin Petzoldt
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
Search for more papers by this authorDr. Manuel Hamburger
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
Search for more papers by this authorProf. Dr. Dirk-Peter Herten
Physikalisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 229, 69120 Heidelberg, Germany
CellNetworks, Single Molecule Spectroscopy, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Richard Wombacher
Institut für Pharmazie und Molekulare Biotechnologie, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany
Search for more papers by this authorProf. Dr. Rasmus R. Schröder
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
CellNetworks, Cryo Electron Microscopy, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorCorresponding Author
Prof. Dr. Uwe H. F. Bunz
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Korwin M. Schelkle
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
These authors contributed equally to this work.
Search for more papers by this authorChristopher Schmid
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
These authors contributed equally to this work.
Search for more papers by this authorM. Sc. Klaus Yserentant
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Physikalisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 229, 69120 Heidelberg, Germany
CellNetworks, Single Molecule Spectroscopy, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorM. Sc. Markus Bender
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Irene Wacker
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Martin Petzoldt
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
Search for more papers by this authorDr. Manuel Hamburger
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
InnovationLab GmbH, Speyerer Strasse 4, 69115 Heidelberg, Germany
Search for more papers by this authorProf. Dr. Dirk-Peter Herten
Physikalisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 229, 69120 Heidelberg, Germany
CellNetworks, Single Molecule Spectroscopy, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorDr. Richard Wombacher
Institut für Pharmazie und Molekulare Biotechnologie, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany
Search for more papers by this authorProf. Dr. Rasmus R. Schröder
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
CellNetworks, Cryo Electron Microscopy, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany
Search for more papers by this authorCorresponding Author
Prof. Dr. Uwe H. F. Bunz
Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany
Center of Advanced Materials, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 225, 69120 Heidelberg, Germany
Search for more papers by this authorGraphical Abstract
Cellfi-Flash: A new fixative for light microscopy of living cells was obtained by functionalizing glutaraldehyde with a photocleavable protecting group. Ester substituents ensure that the masked compound can enter the cell and accumulate therein after esterase-mediated hydrolysis. Incubated cells are instantly fixed and ready for microscopy upon irradiation, whereas conventional aldehyde fixation is diffusion-controlled.
Abstract
Chemical fixation of living cells for microscopy is commonly achieved by crosslinking of intracellular proteins with dialdehydes prior to examination. We herein report a photocleavable protecting group for glutaraldehyde that results in a light-triggered and membrane-permeable fixative, which is nontoxic prior to photocleavage. Lipophilic ester groups allow for diffusion across the cell membrane and intracellular accumulation after enzymatic hydrolysis. Irradiation with UV light releases glutaraldehyde. The in situ generated fixative crosslinks intracellular proteins and preserves and stabilizes the cell so that it is ready for microscopy. In contrast to conventional glutaraldehyde fixation, tissue autofluorescence does not increase after fixation. Caged glutaraldehyde may in future enable functional experiments on living cells under a light microscope in which events of interest can be stopped in spatially confined volumes at defined time points. Samples with individually stopped events could then later be analyzed in ultrastructural studies.
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