Method for generating images with an expanded dynamic range and optical device for carrying out such a method, in particular a laser scanner microscope
Method and apparatus for generating an at least two-dimensional image of at least part of a sample. The method involves scanning the sample. Acquiring at least one light signal by an optoelectronic detector for different areas of the sample. Converting the light signal into an electrical signal. Distributing the electrical signal onto several parallel evaluation channels whose signal evaluations differ from each other so that their dynamic ranges are different. Generating a result signal in each evaluation channel. Selecting at least one of the result signals as a function of one of the result signals in order to generate the image for the sample range concerned. It is also possible to generate one intermediate result signal for each channel, typically from the respective actual result signal and one or more other sources. Thus the signal selection depending on both the result signals and the intermediate result signals are possible.
1. A method for generating an at least two-dimensional image of at least a part of a sample, the method comprising the steps of:
scanning the sample;
acquiring at least one light signal by an optoelectronic detector for different areas of the sample;
converting the at least one light signal into an electrical signal;
distributing the electrical signal onto several parallel evaluation channels whose signal evaluations differ from each other in such a manner that their dynamic ranges are different;
generating a result signal in each evaluation channel; and
selecting at least one of the result signals as a function of one of the result signals of an evaluation channel in order to generate the image for the sample range concerned.
2. The method according to claim 1 , further comprising the steps of generating at least one intermediate result signal of a corresponding evaluation channel, and wherein the selecting step further comprises selecting at least one of the result signals as a function of one of the result signals and/or as a function of an intermediate result signal in order to generate the image for the sample range concerned.
3. The method according to claim 1 , in which each sample range for which a selection is carried out corresponds to a particular image element of the image to be generated.
4. The method according to claim 1 , wherein selecting takes place during scanning.
5. The method according to claim 1 , wherein an evaluation is made for the selection of at least one of the result signals, especially with the determination of a degree of a suitability of the concerned result signal for the generation of the image with expanded dynamic range.
6. The method according to claim 2 , wherein an evaluation is made for the selection of at least one of the result signals and/or of at least one of the intermediate result signals, especially with the determination of a degree of a suitability of the concerned result signal and/or intermediate signal for the generation of the image with expanded dynamic range.
7. The method according to claim 1 , wherein in order to generate the image with a dynamic range that corresponds to a combining of the dynamic ranges of several evaluation channels exactly one scanning pass is carried out.
8. The method according to claim 1 , wherein the selection of the result signal takes place for each sample range using a particular result signal that indicates a photon number.
9. The method according to claim 2 , wherein the selection of the result signal takes place for each sample range using a particular result signal that indicates a photon number, and/or using a particular intermediate result signal that indicates a photon number.
10. The method according to claim 1 , wherein selecting takes place exclusively as a function of one or more of the result signals.
11. The method according to claim 2 , wherein selecting takes place exclusively as a function of one or more of the result signals and/or of one or more of the intermediate result signals.
12. The method according to claim 1 , further comprising the steps of determining one signal-to-noise ratio for each result signal for each sample range; and
selecting and outputting the result signal that has the lowest signal-to-noise ratio.
13. The method according to claim 1 , further comprising the steps of adjusting a gain of the detector or an excitation light intensity of a light source as a function of one of the result signals.
14. The method according to claim 2 , further comprising the steps of adjusting a gain of the detector or an excitation light intensity of a light source as a function of one of the result signals and/or as a function of an intermediate result signal.
15. A control unit that is adjusted for carrying out a method in accordance with claim 1 .
16. An optical device for generating an at least two-dimensional image of at least a part of a sample, the optical device comprising:
a scanning device for scanning the sample;
an optoelectronic detector for acquiring at least one light signal for different areas of the sample and converting the at least one light signal into an electrical signal with several evaluation channels whose signal evaluations differ;
a division circuit for distributing the electrical signal onto several parallel evaluation channels whose signal evaluations differ from each other in such a manner that their dynamic ranges are different;
means for generating a result signal in each evaluation channel; and
means for selecting at least one of the result signals as a function of one of the result signals of an evaluation channel in order to generate the image for the sample range concerned.
17. The device according to claim 16 , further comprising:
means for generating at least one intermediate result signal, and wherein the selecting means further includes means for selecting at least one of the result signals as a function of one of the result signals and/or as a function of an intermediate result signal of an evaluation channel in order to generate the image for the sample range concerned.