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
