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| Flexible Scanning Strategies |  LSM 5 EXCITER |
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| The Digital Signal Processor (DSP) provides a wide variety of possibilities.The DSP controls data acquisition, scanner operation and the acousto-optical tunable filters (AOTFs).
The LSM 5 EXCITER is using a digital signal processor (DSP) to control fast and flexible | |
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| a) | scanners used to position the laser beam within the specimen |
| b) | acusto-optical tunable filter (AOTF) for tuning the power of each single laser line and |
| c) | image acquisition. |
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| Only the exact coordination of these processes enables to program refiued scanning strategies which are necessary to use the LSM for such different methods like for example FRET, FRAP, Photoactivation or Uncaging (Release of caged compounds). Multiple regions of any shape can be selected, scanned with specified laser power and detected with specified settings. | | |
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 | | Bovine endothelium cells; actin filaments (BODIPY FL), mitochondria (MitoTracker Red); some mitochondria are marked (user-definable regions of interest) |
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| Real Region of Interest
The Realtime Electronics guarantees that the specified laser power is achieved for the whole region of interest (ROI) with the accuracy of one pixel.
This ensures, that everything outside will not be exposed and is therefore protected against photo bleaching.
Careful Scanning
This method is also used in at standard image acquisition. Exposure happens during data acquisition only. Therefore the critical point when the scanner turns around and staysfor a moment at one spot gives no cause for concern.
Targeted Bleaching
Real Regious of Interest are a prerequisit for a pixel-wise defined manipulation of samples to bleach, activate or convert fluorescent dyes. | | |
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Bleaching of marked mitochondria with pinpoint accuracy. (left)
Superimposed images of mitochondria before and after bleaching with different channel colors:
Bleached portions are marked in red. (right) |
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