Seal-promoting solutions and pipette perfusion for patch clamping plant cells

Frans J.M. Maathuis, Alison R. Taylor, Sarah M. Assmann, Dale Sanders

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Patch-clamp technology has greatly increased our knowledge of plant membrane transport. However, the success of patch clamping crucially relies on establishing a high resistance (GΩ) seal between the membrane and the patch-clamp pipette. This can prove problematic in many plant-cell preparations. It is therefore of great importance to develop protocols for protoplast isolation, maintenance and seal formation that improve seal rate. This study investigated whether the pH and the K+ and the Cl- concentration of the pipette solution had an effect on the seal formation. High pH and absence of K+ significantly promoted membrane sealing, whereas the concentration of Cl- had no effect. To reap the benefit of seal-promoting pipette solutions and yet retain the option to adjust this solution to experimental requirements, a pipette perfusion apparatus was implemented. The perfusion system was successfully applied in cell-attached patch, excised-patch and whole-cell configurations, using plasma membrane and tonoplast of three different species. The system enables complete solution exchange within minutes end is potentially of great benefit in the study of channel selectivity, the application of (cytoplasmic) channel blockers and the study of primary and secondary transport.

Original languageEnglish (US)
Pages (from-to)891-896
Number of pages6
JournalPlant Journal
Volume11
Issue number4
DOIs
StatePublished - Apr 1 1997

Fingerprint

Plant Cells
Constriction
seals
Perfusion
Membranes
Plant Preparations
Protoplasts
cells
Maintenance
Cell Membrane
tonoplast
Technology
protoplasts
plasma membrane

All Science Journal Classification (ASJC) codes

  • Genetics
  • Plant Science
  • Cell Biology

Cite this

Maathuis, Frans J.M. ; Taylor, Alison R. ; Assmann, Sarah M. ; Sanders, Dale. / Seal-promoting solutions and pipette perfusion for patch clamping plant cells. In: Plant Journal. 1997 ; Vol. 11, No. 4. pp. 891-896.
@article{fbc422a5b62d47cebc9248203acc8dd7,
title = "Seal-promoting solutions and pipette perfusion for patch clamping plant cells",
abstract = "Patch-clamp technology has greatly increased our knowledge of plant membrane transport. However, the success of patch clamping crucially relies on establishing a high resistance (GΩ) seal between the membrane and the patch-clamp pipette. This can prove problematic in many plant-cell preparations. It is therefore of great importance to develop protocols for protoplast isolation, maintenance and seal formation that improve seal rate. This study investigated whether the pH and the K+ and the Cl- concentration of the pipette solution had an effect on the seal formation. High pH and absence of K+ significantly promoted membrane sealing, whereas the concentration of Cl- had no effect. To reap the benefit of seal-promoting pipette solutions and yet retain the option to adjust this solution to experimental requirements, a pipette perfusion apparatus was implemented. The perfusion system was successfully applied in cell-attached patch, excised-patch and whole-cell configurations, using plasma membrane and tonoplast of three different species. The system enables complete solution exchange within minutes end is potentially of great benefit in the study of channel selectivity, the application of (cytoplasmic) channel blockers and the study of primary and secondary transport.",
author = "Maathuis, {Frans J.M.} and Taylor, {Alison R.} and Assmann, {Sarah M.} and Dale Sanders",
year = "1997",
month = "4",
day = "1",
doi = "10.1046/j.1365-313X.1997.11040891.x",
language = "English (US)",
volume = "11",
pages = "891--896",
journal = "Plant Journal",
issn = "0960-7412",
publisher = "Wiley-Blackwell",
number = "4",

}

Seal-promoting solutions and pipette perfusion for patch clamping plant cells. / Maathuis, Frans J.M.; Taylor, Alison R.; Assmann, Sarah M.; Sanders, Dale.

In: Plant Journal, Vol. 11, No. 4, 01.04.1997, p. 891-896.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Seal-promoting solutions and pipette perfusion for patch clamping plant cells

AU - Maathuis, Frans J.M.

AU - Taylor, Alison R.

AU - Assmann, Sarah M.

AU - Sanders, Dale

PY - 1997/4/1

Y1 - 1997/4/1

N2 - Patch-clamp technology has greatly increased our knowledge of plant membrane transport. However, the success of patch clamping crucially relies on establishing a high resistance (GΩ) seal between the membrane and the patch-clamp pipette. This can prove problematic in many plant-cell preparations. It is therefore of great importance to develop protocols for protoplast isolation, maintenance and seal formation that improve seal rate. This study investigated whether the pH and the K+ and the Cl- concentration of the pipette solution had an effect on the seal formation. High pH and absence of K+ significantly promoted membrane sealing, whereas the concentration of Cl- had no effect. To reap the benefit of seal-promoting pipette solutions and yet retain the option to adjust this solution to experimental requirements, a pipette perfusion apparatus was implemented. The perfusion system was successfully applied in cell-attached patch, excised-patch and whole-cell configurations, using plasma membrane and tonoplast of three different species. The system enables complete solution exchange within minutes end is potentially of great benefit in the study of channel selectivity, the application of (cytoplasmic) channel blockers and the study of primary and secondary transport.

AB - Patch-clamp technology has greatly increased our knowledge of plant membrane transport. However, the success of patch clamping crucially relies on establishing a high resistance (GΩ) seal between the membrane and the patch-clamp pipette. This can prove problematic in many plant-cell preparations. It is therefore of great importance to develop protocols for protoplast isolation, maintenance and seal formation that improve seal rate. This study investigated whether the pH and the K+ and the Cl- concentration of the pipette solution had an effect on the seal formation. High pH and absence of K+ significantly promoted membrane sealing, whereas the concentration of Cl- had no effect. To reap the benefit of seal-promoting pipette solutions and yet retain the option to adjust this solution to experimental requirements, a pipette perfusion apparatus was implemented. The perfusion system was successfully applied in cell-attached patch, excised-patch and whole-cell configurations, using plasma membrane and tonoplast of three different species. The system enables complete solution exchange within minutes end is potentially of great benefit in the study of channel selectivity, the application of (cytoplasmic) channel blockers and the study of primary and secondary transport.

UR - http://www.scopus.com/inward/record.url?scp=0031128163&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031128163&partnerID=8YFLogxK

U2 - 10.1046/j.1365-313X.1997.11040891.x

DO - 10.1046/j.1365-313X.1997.11040891.x

M3 - Article

C2 - 9161044

AN - SCOPUS:0031128163

VL - 11

SP - 891

EP - 896

JO - Plant Journal

JF - Plant Journal

SN - 0960-7412

IS - 4

ER -