| |
The principle
The system has been designed for short manipulation times at high laser light yield within the selected sampling area.
You manipulate simultaneously across the complete ROI (region of interest) and find no intensity gradients within the ROI, even with large ROIs. An AOTF (acousto-optical tunable filter) ensures precise laser pulses even in the millisecond range.
Additionally, you capture a brilliant image because of the standard infinity space (laser port).
 The operating principle of DirectFRAP
The laser light is coupled via a multimode fiber into the laser manipulation slider. For optimum adaptation to the individual application case, the diaphragm type can be freely selected immediately before the experiment. There are 19 options with different diaphragm types and sizes available. All points in the ROI are photo manipulated at exactly the same time. |
|
| The operating principle of DirectFRAP |
| |
The laser manipulation slider
The laser manipulation slider is easy to use: simply insert it into the laserport of the Axio Observer microscope. The space required for the complete system is very small. You don't need complex components behind or beside the microscope.
More options: the mask wheel
You are able to use the 19 defined diaphragm positions for dynamical experiments as well as for manipulation of small cell structures. You benefit from fine size graduations for an optimum adaptation to the experiment.
 |
| The diaphragm wheel in the laser manipulation slider provides exceptionally high flexibility. A decisive advantage: if another diaphragm is selected, the laser intensity per unit remains constant. |
| |
The modules Fast Acquisition and Physiology are available for DirectFRAP with the AxioVision system software. In the Physiology module the intensity changes in the ROI are already displayed online during the experiment allowing you to have complete control over your experiment. You can rapidly intervene or optimize the conditions at any time as required.
Laser lines, filter sets and beam combiners
Application | Laser line a) | DirectFRAP b) | Combination
with TIRF 3 | Combination with Cell Observer® SD |
| Bleaching of GFP or Emerald | 488 | FS 74 | FS 74 | Fast beampath switch
or
Beam combiner 80/20 |
| Conversion of Dendra or Eos | 405, 488, 561 | FS 38,
FS 82 | FS 76 |
| Reversible on-/off switching of Dronpa | 405 (on), 488 (off) | FS 83 (on), FS 52 (off) | FS 76 (on), FS 52 (off) |
| Activation / Conversion of PA-GFP / PS-CFP / Phamret | 405, 488 | FS 18 c),
FS 83 | FS 18 c),
FS 76 |
 | |
| a) | Standard laser lines for manipulation are printed in bold. Alternative lasers for manipulation and imaging in system combinations are not in bold |
| b) | An excitation filter wheel is necessary, except for bleaching of GFP or Emerald |
| c) | FS 18 is not necessary for PA-GFP |
| |
| |
|