Laser microsurgery of higher plant cell walls permits patch-clamp access

Gordon H. Henriksen, Alison R. Taylor, Colin Brownlee, Sarah M. Assmann

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Plasma membranes of guard cells in epidermal peels of Vicia faba and Commelina communis can be made accessible to a patch-clamp pipet by removing a small portion (1-3 μm in diameter) of the guard cell wall using a microbeam of ultraviolet light generated by a nitrogen laser. Using this laser microsurgical technique, we have measured channel activity across plasma membranes of V. faba guard cells in both cell-attached and isolated patch configurations. Measurements made in the inside-out patch configuration revealed two distinct K+-selective channels. Major advantages of the laser microsurgical technique include the avoidance of enzymatic protoplast isolation, the ability to study cell types that have been difficult to isolate as protoplasts or for which enzymatic isolation protocols result in protoplasts not amenable to patch-clamp studies, the maintenance of positional information in single-channel measurements, reduced disruption of cell-wall-mediated signaling pathways, and the ability to investigate intercellular signaling through studies of cells remaining situated within tissue.

Original languageEnglish (US)
Pages (from-to)1063-1068
Number of pages6
JournalPlant physiology
Volume110
Issue number4
DOIs
StatePublished - Apr 1996

Fingerprint

Microsurgery
guard cells
Plant Cells
Cell Wall
protoplasts
lasers
Lasers
cell walls
Protoplasts
Vicia faba
plasma membrane
Commelina communis
cells
Commelina
ultraviolet radiation
Cell Membrane
Gas Lasers
Ultraviolet Rays
nitrogen
methodology

All Science Journal Classification (ASJC) codes

  • Physiology
  • Genetics
  • Plant Science

Cite this

Henriksen, Gordon H. ; Taylor, Alison R. ; Brownlee, Colin ; Assmann, Sarah M. / Laser microsurgery of higher plant cell walls permits patch-clamp access. In: Plant physiology. 1996 ; Vol. 110, No. 4. pp. 1063-1068.
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Laser microsurgery of higher plant cell walls permits patch-clamp access. / Henriksen, Gordon H.; Taylor, Alison R.; Brownlee, Colin; Assmann, Sarah M.

In: Plant physiology, Vol. 110, No. 4, 04.1996, p. 1063-1068.

Research output: Contribution to journalArticle

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AU - Henriksen, Gordon H.

AU - Taylor, Alison R.

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AB - Plasma membranes of guard cells in epidermal peels of Vicia faba and Commelina communis can be made accessible to a patch-clamp pipet by removing a small portion (1-3 μm in diameter) of the guard cell wall using a microbeam of ultraviolet light generated by a nitrogen laser. Using this laser microsurgical technique, we have measured channel activity across plasma membranes of V. faba guard cells in both cell-attached and isolated patch configurations. Measurements made in the inside-out patch configuration revealed two distinct K+-selective channels. Major advantages of the laser microsurgical technique include the avoidance of enzymatic protoplast isolation, the ability to study cell types that have been difficult to isolate as protoplasts or for which enzymatic isolation protocols result in protoplasts not amenable to patch-clamp studies, the maintenance of positional information in single-channel measurements, reduced disruption of cell-wall-mediated signaling pathways, and the ability to investigate intercellular signaling through studies of cells remaining situated within tissue.

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