A simple device to immobilize protists for electrophysiology and microinjection

Author:

Kulkarni Anirudh1ORCID,Elices Irene1,Escoubet Nicolas2,Pontani Léa-Laetitia2ORCID,Prevost Alexis Michel2ORCID,Brette Romain1ORCID

Affiliation:

1. Sorbonne Université, INSERM, CNRS, Institut de la Vision, 17 rue Moreau, F-75012 Paris, France

2. Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS), Laboratoire Jean Perrin (LJP), F-75005, Paris, France

Abstract

ABSTRACT We present a simple device to mechanically immobilize motile cells such as ciliates. It can be used in particular for intracellular electrophysiology and microinjection. A transparent filter with holes smaller than the specimen is stretched over an outlet. A flow is induced by either a peristaltic pump or a depressurized tank, mechanically entraining cells to the bottom, where they are immobilized against the filter. The cells start swimming again as soon as the flow is stopped. We demonstrate the device by recording action potentials in Paramecium and injecting a fluorescent dye into the cytosol.

Funder

Centre National de la Recherche Scientifique

Sorbonne Université

Agence Nationale de la Recherche

Publisher

The Company of Biologists

Subject

Insect Science,Molecular Biology,Animal Science and Zoology,Aquatic Science,Physiology,Ecology, Evolution, Behavior and Systematics

Reference27 articles.

1. DNA microinjection into the macronucleus of paramecium;Beisson;Cold Spring Harb. Protoc.,2010

2. The physiological basis of taxes in paramecium;Kung;Annu. Rev. Physiol.,1982

3. Bioelectric control of ciliary activity;Eckert;Science,1972

4. Ionic mechanisms of excitation in Paramecium;Eckert;Annu. Rev. Biophys. Bioeng.,1979

5. Paramecium

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