Dentritic Patch Clamp Recording - Femtonics

12 downloads 75 Views 584KB Size Report
Guide points can be positioned by the user according to the scanned transmitted or two-photon images. • Only a simple calibration step is required before using ...
Femto2D MULTIPHOTON LASER SCANNING MICROSCOPE

Dentritic Patch Clamp Recording „Imaging the speed of life”

© Femtonics Kft. Tűzoltó u. 59. H-1094 Budapest Hungary

www.femtonics.eu [email protected] +36-1-2103349

Dendritic patch-clamp recording is supported by our simultaneous oblique illumination and two-photon laser scanning microscopy Description Our infrared laser scanning method provides an excellent, high quality real-time image of the fine tip of the high resistance patch-pipette located deep in the tissue. In combination with simultaneous two-photon imaging, this provides an excellent tool for dendritic patch clamp recording. The location of dendritic patch clamp recording can be selected in advance by the user using two-photon imaging data. Based on this input, an automatic software module (Luigsneumann1 module, Femtonics Ltd.) moves the tip of the pipette to the selected dendritic location.

Advantages • Guide points can be positioned by the user according to the scanned transmitted or two-photon images • Only a simple calibration step is required before using a new recording pipette • The recording pipettes are moved in the tissue only diagonally in order to minimize tissue damage • Pipette movement and distance to target location can be monitored in realtime • The final steps of dendritic targeting can be controlled both manually and automatically • Real-time transmitted infrared and two photon images can be shown separately or in overlay • Fast water cooled scanners provide video-rate imaging of two-photon and transmitted infrared data Patch Clamp mode CCD camera image using oblique IR illumination. The „bubble” helping the creation of the gigaohm resistance is well visible

A

20µm

20µm

copyright 2011 © Femtonics Kft. [email protected] | Tűzoltó u. 59. H-1094 Budapest, Hungary | +36-1-2103349 | www.femtonics.eu

Experiments made by Femto2D microscope Dendritic patch-clamping with real-time laser scanning Simultaneous real-time visualization of the patch pipette and the dendrite in order to create dendritic patch-clamp recording from thin dendrites

B

tIR

tIR+2P

2P

20µm

20µm

20µm

2P

tIR

20µm

tIR+2P

20µm

20µm

Further example images Two-photon guided dendritic patch clamping using different LUT settings for visualisation

C 20µm

50µm 20µm

50µm 50µm

Measurementss

performed in cooperation with the IEM-HAS, Budapest, Hungary

copyright 2011 © Femtonics Kft. [email protected] | Tűzoltó u. 59. H-1094 Budapest, Hungary | +36-1-2103349 | www.femtonics.eu