DEVICE AND METHOD FOR RASTER-SCAN OPTOACOUSTIC IMAGING

Abstract

The invention relates to a device and corresponding method for raster-scan optoacoustic imaging, the device comprising: a radiation source comprising at least one Raman laser source, the radiation source being configured to generate a plurality of pulses of electromagnetic radiation, each of the pulses comprising portions of electromagnetic radiation at two or more distinct wavelengths, and at least one acousto-optic tunable filter configured to select, from at least one of the pulses, one of the portions of electromagnetic radiation at one of the wavelengths; an irradiation unit configured to irradiate a region of interest of an object, in particular a biological tissue, with the selected portion of electromagnetic radiation of the at least one pulse; a detection unit configured to detect acoustic waves emitted from the region of interest in response to irradiating the region of interest with the selected portion of electromagnetic radiation of the at least one pulse; and a scanning unit configured to move the irradiation unit and detection unit, on the one hand, and/or the region of interest, on the other hand, along at least one dimension relative to each other so as to position the irradiation unit and detection unit at a plurality of different locations along the at least one dimension relative to the region of interest, and to control the detection unit to detect the acoustic waves at the plurality of locations.

Claims

1. A device for raster-scan optoacoustic imaging, the device comprising: a radiation source comprising at least one Raman laser source, the radiation source being configured to generate a plurality of pulses of electromagnetic radiation, each of the pulses comprising portions of electromagnetic radiation at two or more distinct wavelengths, and at least one acousto-optic tunable filter configured to select, from at least one of the pulses, one of the portions of electromagnetic radiation at one of the wavelengths, an irradiation unit configured to irradiate a region of interest of an object, in particular a biological tissue, with the selected portion of electromagnetic radiation of the at least one pulse, a detection unit configured to detect acoustic waves emitted from the region of interest in response to irradiating the region of interest with the selected portion of electromagnetic radiation of the at least one pulse, and a scanning unit configured to move the irradiation unit and detection unit, on the one hand, and/or the region of interest, on the other hand, along at least one dimension relative to each other so as to position the irradiation unit and detection unit at a plurality of different locations along the at least one dimension relative to the region of interest, and to control the detection unit to detect the acoustic waves at the plurality of locations.

2. The device according to claim 1, the acousto-optic tunable filter being configured to select, from each of at least two of the pulses, in particular from each of at least two subsequent pulses, one of the portions of electromagnetic radiation at different wavelengths, the irradiation unit being configured to irradiate the region of interest with the selected portions of electromagnetic radiation of the at least two, in particular subsequent, pulses and the scanning unit being configured to control the detection unit to detect acoustic waves emitted from the region of interest in response to irradiating the region of interest with the selected portions of electromagnetic radiation of the at least two, in particular subsequent, pulses.

3. The device according to claim 2, the scanning unit being configured to control the detection unit to detect acoustic waves emitted from the region of interest in response to irradiating the region of interest with the selected portions of electromagnetic radiation of the at least two, in particular subsequent, pulses while continuously moving the irradiation unit and detection unit, on the one hand, and the region of interest, on the other hand, relative to each other.

4. The device according to claim 1, the Raman laser source comprising a pump laser, in particular a solid-state laser, configured to emit a plurality of pulses of first radiation at a first wavelength and/or the radiation source further comprising at least one second harmonic generator configured to convert a portion of the first radiation at the first wavelength to a second radiation at a second wavelength, wherein the second wavelength equals to half of the first wavelength.

5. The device according to claim 4, the Raman laser source comprising an active medium, in particular a glass fiber or a crystal or a gas cell, configured to emit, in response to a stimulation by the first radiation and/or second radiation, a plurality of pulses of Raman radiation, each of the pulses comprising one or more portions of electromagnetic radiation at one or more distinct Raman wavelengths.

6. The device according to claim 5, the radiation source further comprising at least one extracting unit configured to extract a portion of the first radiation and/or a portion of the second radiation prior to stimulating the active medium so as to bypass the active medium and an alignment unit configured to co-align the extracted portion of the first radiation and/or second radiation with the Raman radiation.

7. The device according to claim 5, each of the pulses of Raman radiation emitted by the active medium comprising, in addition to the one or more portions of electromagnetic radiation at the one or more distinct Raman wavelengths, a residual portion of the first radiation at the first wavelength and/or a residual portion of the second radiation at the second wavelength and/or at least one residual portion of electromagnetic radiation at the one or more distinct Raman wavelengths.

8. The device according to claim 7, the residual portion of the first radiation at the first wavelength and/or the residual portion of the second radiation at the second wavelength and/or the at least one residual portion of electromagnetic radiation at the one or more distinct Raman wavelengths being co-aligned with the Raman radiation.

9. The device according to claim 5, wherein each of the pulses of the plurality of pulses of electromagnetic radiation generated by the radiation source comprises Raman radiation at the one or more distinct Raman wavelengths.

10. The device according to claim 9, wherein each of the pulses of the plurality of pulses of electromagnetic radiation generated by the radiation source further comprises the extracted or residual portion of the first radiation at the first wavelength and/or the extracted or residual portion of the second radiation at the second wavelength.

11. The device according to claim 5, wherein the active medium exhibits different Raman shifts along different directions of propagation of the first radiation or of the second radiation within the active medium.

12. The device according to claim 11, wherein the active medium is a crystalline material exhibiting a crystallographic axis and the polarization of the first radiation or of the second radiation is controllable and/or controlled so as to select a desired Raman shift by virtue of the orientation of said polarization relative to the crystallographic axis of the active medium.

13. The device according to claim 5, wherein the polarization of the Raman radiation and/or of the first radiation and/or of the second radiation is controllable and/or controlled so as to align the Raman radiation and/or the extracted or residual portion of the first radiation and/or the extracted or residual portion of the second radiation to the at least one acousto-optic tunable filter to maximize the efficiency of the at least one acousto-optic tunable filter.

14. The device according to claim 1, wherein the detection unit comprises a focused ultrasound transducer configured to detect the acoustic waves emitted from the region of interest and/or the irradiation unit is configured to irradiate the region of interest with a divergent beam of the selected portion of electromagnetic radiation of the at least one pulse.

15. A method for raster-scan optoacoustic imaging comprising the following steps: generating a plurality of pulses of electromagnetic radiation by means of a radiation source comprising at least one Raman laser source, each of the pulses comprising portions of electromagnetic radiation at two or more distinct wavelengths, selecting, from at least one of the pulses, one of the portions of electromagnetic radiation at one of the wavelengths by means of at least one acousto-optic tunable filter, irradiating a region of interest of an object, in particular a biological tissue, with the selected portion of electromagnetic radiation of the at least one pulse by means of an irradiation unit, detecting acoustic waves emitted from the region of interest in response to irradiating the region of interest with the selected portion of electromagnetic radiation of the at least one pulse by means of a detection unit, and moving the irradiation unit and detection unit, on the one hand, and/or the region of interest, on the other hand, along at least one dimension relative to each other so as to position the irradiation unit and detection unit at a plurality of different locations along the at least one dimension relative to the region of interest, and detecting acoustic waves at the plurality of locations.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Further additional or alternative aspects, advantages, features and examples of the present invention will be apparent from the following description of following figures:

[0037] FIG. 1 shows an example of a device for raster-scan optoacoustic imaging;

[0038] FIG. 2 shows a first and second example of a radiation source;

[0039] FIG. 3 shows a third and fourth example of a radiation source;

[0040] FIG. 4 shows an example of an acousto-optic tunable filter; and

[0041] FIG. 5 shows a table with examples of wavelengths of the electromagnetic radiation generated by the radiation source.

[0042] FIG. 1 shows an example of a device for raster-scan optoacoustic imaging comprising a radiation source 10 which is configured to generate a plurality of pulses P of electromagnetic radiation at distinct wavelengths or wavelength ranges. In present example, the radiation source 10 is configured to generate sequences of five pulses P containing electromagnetic radiation at five distinct wavelengths.

[0043] The pulses P are coupled into a proximal end of an optical guiding element 20, e.g. a single fiber, multiple fibers or a fiber bundle, which is configured to guide the pulses P towards an object 1 to be imaged and to irradiate a region of interest of the object 1 with the pulses. In present example, the guiding element 20 is fed, as indicated by dashed lines, through a probe 30 such that a divergent beam of electromagnetic radiation of the pulses P emerging from a distal end of the guiding element 20 impinges on the object 1.

[0044] The probe 30 comprises a detection unit 31 which is configured to detect acoustic waves emitted from the region of interest of the object 1 in response to an irradiation with the pulses P. Preferably, the detection unit 31 comprises a focused single-element ultrasound transducer exhibiting a focus point and/or focal zone and being configured to detect ultrasound waves within a conically shaped sensitivity field of the transducer.

[0045] The probe 30 including the detection unit 31 and the distal end of the guiding element 20 is coupled to a scanning unit 40 which is configured to move the probe 30 relative to the object 1 in x- and y-direction to position the probe 30 at a plurality of different locations (x, y) in the x-y plane. In present example, the different locations (x, y) of the probe 30 relative to the object 1 are illustrated by a plurality of dots depicted over the region of interest of the object 1. Preferably, the dots constitute a grid of a plurality of positions (x, y) over which the detection unit 31, in particular the single-element transducer, is located, in particular centered, at the time when respective pulses P are generated and/or impinge on the region of interest. In present example, the probe 30 and/or detection unit 31 is moved relative to the object 1 along a meander-like or meandering trace. Optionally, the scanning unit 40 may be configured to move the probe 30 also in z-direction relative to the object 1.

[0046] Preferably, a control unit 70, for example a computer system, is provided for controlling the irradiation unit 10 and/or the scanning unit 40 and/or the detection unit 31 accordingly. Preferably, an amplifier 50 is provided for amplifying the signals corresponding to the acoustic waves detected by the detection unit 31. Preferably, a data acquisition card 60 (DAQ card) is provided via which the amplified signals are forwarded to the control unit 70 for processing and/or image reconstruction and/or multispectral processing (e.g. unmixing or other operations between images of different wavelength, e.g. division, subtraction, overlay, etc) and/or image display.

[0047] Preferably, the detection unit 31 detects acoustic waves emitted from the region of interest in response to subsequently irradiating the region of interest with the selected portions P of electromagnetic radiation at different wavelengths of subsequent pulses while continuously moving the probe 30 including the distal end of the irradiation unit 20 and the detection unit 31 relative to the object 1. That is, the scanning motion of the probe 30 relative to the object 1 advantageously continues while the object is subsequently irradiated with pulses of electromagnetic radiation at the selected different wavelengths, in present example five different wavelengths.

[0048] In particular when providing high pulse repetition rates of preferably more than 1 kHz, which are advantageously enabled by a radiation source 10 combining a Raman laser source and an acousto-optic tunable filter as will be explained in more detail below with reference to FIGS. 2 to 4, the time elapsing during irradiating the object 1 with subsequent pulses P at the different wavelengths is very short so that a possible motion of a living object, e.g. an organ or body part of a human or animal, during this time has no or only a negligible effect on the image quality, so that in particular motion artifacts can be avoided or at least significantly reduced. For example, at a pulse repetition rate of approx. 1.4 kHz the time required for irradiating the object with four subsequent pulses at four different wavelengths is approx. 1/1.4 kHz×4=0.7 ms×4=2.8 ms which is considerably shorter than the time required for a typical physiological movement, e.g. approx. 0.8 s for heart beat.

[0049] FIG. 2 shows a first and second example of a radiation source, which is preferably used as the radiation source 10 in the device shown in FIG. 1.

[0050] In the first example shown in the upper part of FIG. 2, a pump laser 11, e.g. a solid-state laser, is provided to generate a plurality of pulses of first radiation at a first wavelength, e.g. at 1064 nm or 1030 nm. Further, a second harmonic generator 12 is provided which converts the first radiation at the first wavelength to a second radiation at a second wavelength, e.g. 532 nm or 515 nm, respectively, which equals half the first wavelength.

[0051] A first portion of the pulsed second radiation at the second wavelength is guided to an active medium 13, e.g. a glass fiber, crystal or gas cell, which emits, due to Raman scattering stimulated by the second radiation, a plurality of pulses of Raman radiation at different distinct wavelengths. In present example, as indicated by three differently dashed lines, pulsed Raman radiation at in total three different distinct wavelengths is generated.

[0052] In general, wavelengths of the Raman radiation (also referred to as Raman wavelengths) given in the context of present disclosure may vary by ±2 nm.

[0053] Preferably, an extracting unit 15 is provided which is configured to transmit the first portion of the pulsed second radiation at the second wavelength to the active medium 13 and to extract a second portion (indicated by dashed-dotted line) of the pulsed second radiation prior to reaching the active medium 13 so as to bypass the active medium 13 and to be aligned, preferably by means of an alignment unit 16, with the pulsed Raman radiation emerging from the active medium 13. In this way, in present example, a plurality of pulses of electromagnetic radiation each comprising in total four portions of electromagnetic radiation at four different wavelengths, e.g. 532 nm (bypassed second radiation) and 555 nm, 579 nm and 606 nm (Raman radiation), is obtained.

[0054] Further, an acousto-optic filter 14 is provided which is configured to select from each of the obtained pulses of electromagnetic radiation one out of four portions of electromagnetic radiation at one of the four distinct wavelengths.

[0055] Preferably, the acousto-optic filter 14 is configured to deflect the different portions of electromagnetic radiation depending on the respective wavelength and to be controlled to deflect the portions of electromagnetic radiation such that only a pre-defined portion (i.e. the portion to be selected) passes an aperture 17, while the other portions are blocked.

[0056] Preferably, the acousto-optic filter 14 is configured to deflect portions of electromagnetic radiation at different wavelengths so quickly that the different portions contained in subsequent pulses can be subsequently selected, which is also referred as “per-pulse tuning” in connection with present disclosure. In this way, in present example, a first portion of electromagnetic radiation at a first wavelength, e.g. 532 nm, of a first pulse passes the aperture 17, a second portion of electromagnetic radiation at a second wavelength, e.g. 555 nm, of a subsequent second pulse passes the aperture 17, and so on.

[0057] In the second example shown in the lower part of FIG. 2, a first portion of a plurality of pulses of first radiation at a first wavelength, e.g. at 1064 nm or 1030 nm, generated by pump laser 11 is transmitted to the active medium 13 which generates a plurality of pulses of Raman radiation at distinct wavelengths, e.g. at in total three different distinct wavelengths (as indicated by three differently dashed lines).

[0058] Preferably, an extracting unit 15 is provided which is configured to transmit the first portion of the pulsed first radiation at the first wavelength to the active medium 13 and to extract a second portion of the pulsed first radiation prior to reaching the active medium 13 so as to bypass the active medium 13 and to be aligned, preferably by means of alignment unit 16, with the pulsed Raman radiation emerging from the active medium 13. In this way, in present example, a plurality of pulses of electromagnetic radiation each comprising in total four portions of electromagnetic radiation at four different wavelengths, e.g. 1064 nm (bypassed first radiation) and e.g. 1159 nm, 1272 nm and 1410 nm (Raman radiation), is obtained.

[0059] The second harmonic generator 12 converts the portions of electromagnetic radiation at the different (initial) wavelengths contained in each pulse to portions of electromagnetic radiation at different wavelengths corresponding to respectively half the (initial) wavelengths, e.g. 532 nm, 579 nm, 636 nm and 705 nm, respectively.

[0060] From each of the pulses of electromagnetic radiation obtained in this way, one out of the four portions of electromagnetic radiation at one of the four distinct wavelengths is selected by means of acousto-optic filter 14. The above explanations regarding the opto-acoustic filter 14 and the aperture 17 apply accordingly.

[0061] For further details on the second example of the radiation source, the above explanations given with reference to the first example of the radiation source apply accordingly.

[0062] FIG. 3 shows a third and fourth example of a radiation source, which is preferably used as the radiation source 10 in the device shown in FIG. 1. Basically, the above explanations with reference to the first and second example of the radiation source apply accordingly to the third and fourth example, respectively.

[0063] In contrast to the first and second example, however, no extraction unit 15 and/or bypassing is used to combine a portion of pulsed pump radiation (i.e. second portion of pulsed second radiation in the first example, second portion of pulsed first radiation in the second example) with the portions pulsed Raman radiation emerging from the active medium 13.

[0064] Rather, in the third example, from the second radiation (indicated by dashed dotted line) emerging from the second harmonic generator 12 and coupled into the active medium 13, a residual portion R is not converted into Raman radiation and emerges from the active medium 13. The residual portion R is intrinsically co-aligned with the different portions of stimulated Raman radiation (indicated by differently dashed lines) so that an additional alignment unit 16 (see FIG. 2) is dispensable. The above explanations regarding the opto-acoustic filter 14 and the aperture 17 apply accordingly.

[0065] Likewise, in the fourth example, from the first radiation (indicated by solid line) emerging from the pump laser 11 and coupled into the active medium 13, a residual portion R is not converted into Raman radiation and emerges from the active medium 13 in a co-aligned manner with the different portions of stimulated Raman radiation (indicated by differently dashed lines) so that an additional alignment unit 16 (see FIG. 2) is dispensable. The above explanations regarding the second harmonic generator 12, the opto-acoustic filter 14 and the aperture 17 apply accordingly.

[0066] FIG. 4 shows an example of an acousto-optic tunable filter, which is preferably used as the acousto-optic filter 14 in the radiation source shown in FIGS. 2 and 3.

[0067] Preferably, the opto-acoustic filter, which may also be referred to as acousto-optic modulator (AOM) or Bragg cell, uses the acousto-optic effect to diffract the pulsed electromagnetic radiation containing several portions at different wavelengths (in present example 532 nm, 555 nm, 579 nm and 606 nm) using sound waves generated by a piezoelectric transducer 14a attached to a material 14b such as quartz or glass. An acoustic absorber 14c is provided at a side of the material 14b opposite to the transducer 14a. An oscillating electric signal drives the transducer 14c to vibrate, which creates sound waves in the material 14b which act as moving periodic planes of expansion and compression that change the index of refraction of the material 14b. The incoming electromagnetic radiation scatters off the resulting periodic index modulation and interference occurs so that the portions of electromagnetic radiation at different wavelengths are split up as exemplarily shown in the Figure.

[0068] Further preferred or alternative aspects of present disclosure are discussed in the following.

[0069] The disclosed device and method provide a fast tuning multispectral RSOM scanner using a Raman laser source in combination with an acousto-optic tunable filter (AOTF), which reaches pulse repetition rates of more than 1 kHz and a per-pulse tunability of the wavelength. This allows for fast multispectral raster scanning with intrinsic co-registration of the 3D volume between different wavelengths.

[0070] Preferably, the preferably focused single-element ultrasound transducer has a central frequency in the range from 10 to 100 MHz and/or a bandwidth of more than 50% of the respective central frequency.

[0071] Preferably, 3D images can be reconstructed based on the acoustic waves detected separately for each wavelength.

[0072] Further spectral processing is preferred to visualize the imaged absorbers, using spectral information from the different wavelengths.

[0073] Preferably, the pump laser 11 is a solid state laser, e.g. 1064 nm (Nd:YAG) or 1030 nm (Yb:YAG), with a pulse length in the range from 200 ps to 10 ns.

[0074] Preferably, the active material 13 provides a set of Raman wavelengths in the visible range, e.g. but not limited to 555 nm, 579 nm, 606 nm, 635 nm. Preferably, the set of Raman wavelengths is chosen depending on the orientation of the polarization relative to the medium (crystal).

[0075] Preferably, the set of Raman wavelengths is fine-tuned by the pump wavelength, Raman active material, polarization, and second harmonic generation to get wavelengths that fit well to the spectra of absorbers to be distinguished, e.g. 532-555-579-606 nm to discriminate between melanin, oxyhemoglobin, deoxyhemoglobin.

[0076] Preferably, the tuning speed of the AOTF can reach more than 1 kHz. Due to non-perfect filtering of the AOTF, residual energies from the other wavelengths may be present. Preferably, the remaining wavelengths get damped by output aperture 17. Preferably, the set of wavelengths gets continuously repeated during the scan.

[0077] Preferably, spectral processing scheme of the different wavelength images is performed using at least one of the following methods: [0078] Linear regression making use of known absorption spectra, e.g. spectra of melanin, oxyhemoglobin, deoxyhemoglobin, etc. [0079] ICA, in particular guided ICA [0080] Ratios [0081] Addition or subtraction [0082] Color/Channel mixing.

[0083] Preferably, present disclosure relates to a multispectral raster-scanning optoacoustic imaging device and method, which uses laser pulses of different wavelengths to excite optical absorbers within a sample, and a single element focused ultrasound detector to capture the ultrasound signal emitted by the exited absorbers following energy conversion due to the photoacoustic effect.

[0084] According to a preferred implementation, the irradiation unit, in particular the distal end of optical guiding element 20, is co-axially arranged with an axis of the detection unit 31 and/or the probe 30 and guided through the detection unit 31 and/or probe 30 to enable a co-axial irradiation and detection.

[0085] Preferably, the Raman laser medium 13 is pumped by a solid-state laser, e.g. with 1064 nm or 1030 nm, with a pulse length of shorter than 10 ns, preferably shorter than 5 ns.

[0086] Preferably, second harmonic generation can be performed either directly on the pump wavelength or after the Raman active medium 13 (crystal). A portion of the pump wavelength or frequency-doubled pump wavelength may be extracted and co-aligned after the Raman medium 13, while the major energy passes through the Raman medium 13 producing a set of Stokes-shifted wavelengths depending on the pump wavelength. By choosing the polarization of the pump beam different sets of Raman wavelengths can be selected (linear or rotate polarization).

[0087] Preferably, at the laser outlet an acousto-optic-tunable filter is provided to filter out one of the Raman wavelengths and/or the frequency-doubled pump wavelength.

[0088] Preferably, the set of Raman wavelengths is chosen depending on the orientation of the polarization relative to the active medium 13 (crystal). Preferably, the wavelengths 555 nm, 579 nm and 606 nm originate from the rotate polarization.

[0089] Preferably, the pulse length of pump and Raman radiation is in the range of 2 to 3 ns. All of the Raman wavelengths are intrinsically co-aligned. In addition, a fraction of the pump wavelength can be extracted before the Raman crystal, and co-aligned with the Raman wavelengths before the AOTF.

[0090] Preferably, an AOTF driver is used to generate a sequence of signals, preferably electric signals provided to the piezoelectric transducer 14a at the medium 14b of the AOTF, to filter out a single wavelength at the laser output, while the rest of the wavelengths is damped inside the laser housing.

[0091] Due to not-perfect filtering of the AOTF (side-lobes), residual portions from neighboring wavelengths are present in addition to (main) portions at the different wavelengths. Preferably, this may be considered in the unmixing scheme.

[0092] In the following, examples of approximate main and residual portions at different wavelengths are given: [0093] R1: 97% 532 nm+3% 555 nm [0094] R2: 94% 555 nm+3% 532 nm+3% 579 nm [0095] R3: 85% 579 nm+9% 555 nm+6% 606 nm [0096] R4: 94% 606 nm+6% 579 nm

[0097] Preferably, the laser energies are calibrated before unmixing procedures are performed.

[0098] FIG. 5 shows a table with examples of wavelengths (in units of [nm]) of electromagnetic radiation generated by a radiation source 10 comprising Potassium Gadolinium Tungstate KGd(WO.sub.4).sub.2, also referred to as KGW, exhibiting a Stokes shift of 768 cm.sup.−1 or 901 cm.sup.−1, or Barium Nitrate Ba(NO.sub.3).sub.2 exhibiting a Stokes shift of 1048 cm.sup.−1 as the active medium 13 (see FIGS. 2 and 3) for different pump wavelengths (“Pump”).

[0099] In the upper part of the table, the pump wavelengths of 532 nm and 515 nm correspond to second wavelengths of second radiation within the meaning of present disclosure and were obtained by converting first radiation generated by pump laser 11 at 1064 nm or 1030 nm, respectively, with a second harmonic generator 12 (see upper part of FIGS. 2 and 3 and respective description). Dependent on the pump wavelength and the material of the active medium 13 of the Raman source, Raman radiation at different Raman wavelengths “1st Raman” to “4th Raman” is obtained.

[0100] In the lower part of the table, the pump wavelengths of 1064 nm and 1030 nm correspond to first wavelengths of first radiation within the meaning of present disclosure which was generated by pump laser 11 at 1064 nm or 1030 nm, respectively. Dependent on the pump wavelength and the material of the active medium 13 of the Raman source, Raman radiation at different Raman wavelengths “1st Raman” to “4th Raman” is obtained. In case of a subsequent conversion of the obtained radiation (Pump and Raman) with a second harmonic generator 12 (see lower part of FIGS. 2 and 3 and respective description), radiation at half the wavelength “Pump+SHG” and “1st Raman+SHG” to “4th Raman+SHG” is obtained in each case.

[0101] In the table, preferably used sets of distinct wavelengths (“Pump”, “Pump+SHG”, “Raman”, “Raman+SHG”) comprising four (upper part of table) or, respectively, three (lower part of table) distinct wavelengths are highlighted by a shaded background. The highlighted sets of distinct wavelengths are particularly advantageous for obtaining multispectral raster-scan optoacoustic images with high informative and/or diagnostic value.