Optical Radiation Detection Device

Description

[0001] The invention relates to a method and a device for the optical in vitro detection of movement in a biological sample and / or for the optical in vitro detection of movement of a sample component of the biological sample, wherein the diameter of the biological sample is preferably at least 100 µm.

[0002] Practical experience has shown that there is a need to monitor the active dynamics of closed and three-dimensional cell and tissue cultures in areas such as developmental biology, toxicity testing, and pharmaceutical research.

[0003] For example, tissue samples grown from embryonic stem cells that have differentiated into muscle tissue are used in toxicity tests to assess the harmfulness of a substance under investigation. This involves examining whether a substance applied to the muscle tissue affects muscle contractions, which can be an indicator of the substance's toxicity. This requires measurement techniques to detect movement, such as contraction, within such a three-dimensional biological sample in the form of a cell cluster. Typical diameters of these cell clusters range from 50 to 400 µm, although diameters in the millimeter range are also possible.

[0004] These studies are currently conducted primarily through visual observations and only rarely through video microscopy with subsequent image analysis. The former are time-consuming and always involve subjective assessment. The latter has the disadvantage of requiring complex imaging optics and sophisticated image analysis, coupled with considerable computational effort. Furthermore, its inherent sensitivity to minor shifts is a disadvantage.

[0005] Non-optical methods such as impedance measurements only work in contact with the sample. However, if the sample shape deviates from a plane (adherent monolayer) or is even three-dimensional, and is also freely floating in a medium, the aforementioned techniques are not applicable.

[0006] Automated imaging methods have the disadvantage that, for example, with a depth of field of 10 µm and the aforementioned typical cell cluster size of 50 to 400 µm, 5 to 40 image planes of the sample would have to be measured. With a typical minimum measurement time of approximately 10 seconds to detect movement, and the additional time required for repositioning or refocusing, such methods are not suitable for quickly monitoring a large number of samples.

[0007] Serial measurement of large numbers of samples is generally not advisable due to the relatively long observation times inherent in the timescales of biological dynamics. In practice, however, there is a need to perform such motion detection on a large number of separated samples, for example, in a multiwell plate, also known as a microtiter plate, with 96 or 384 wells. Imaging techniques are unsuitable for the parallel measurement of a large number of such samples because, for geometric and spatial reasons, it is difficult to position a conventional imaging optical device at each well of the multiwell plate.

[0008] A measurement system for detecting the motility of microorganisms is known from the publication Pomarico J et al: "Compact device for assessment of microorganism motility", Review of Scientific Instruments, AIP, Melville, NY, US, Vol. 75, No. 11, (2005-11-01), pages 4727-4731. The approach is based on dynamic speckle interferometry, whereby a speckle pattern is recorded using a CCD camera and subsequently evaluated using a PC and image processing software.

[0009] It is therefore an object of the invention to provide an improved method for detecting motion in a biological sample and / or motion of a sample component of the biological sample, thereby avoiding the disadvantages of conventional techniques. In particular, the invention aims to provide a robust, non-contact method for motion detection that does not require complex image analysis. A further object of the invention is to provide a method suitable for the parallel analysis of a large number of samples in a screening environment. Another object is to provide a device for detecting motion in a spatially extended biological sample that avoids the disadvantages of conventional devices.

[0010] These problems are solved by devices and methods with the features of the independent claims. Advantageous embodiments and applications of the invention are described in the dependent claims and are explained in more detail in the following description with partial reference to the figures.

[0011] According to a first aspect of the invention, a method for the optical in vitro detection of movement in a biological sample and / or movement of a sample component of the biological sample is provided. The biological sample is a biological sample with spatial extension. The biological sample is preferably a biological sample comprising a plurality, in particular a multiplicity, of biological cells. The diameter of the biological sample can be at least 50 micrometers (µm) in at least one spatial direction, more preferably in all spatial directions, and is often more than 100 µm in all spatial directions. Another embodiment provides that the diameter of the biological sample is larger than the diffraction-limited resolution achievable by the optical wide-field illumination device. However, the invention is not limited to biological samples with such dimensions.

[0012] The biological sample can be a three-dimensional cell and / or tissue culture, e.g., in the form of a cell cluster. The biological sample can specifically include living cells, such as muscle cells, or more generally, cells that exhibit active dynamics and / or can trigger movement. The detection of movement in the biological sample refers specifically to movement within the sample itself or movement of a component of the biological sample. In other words, the aim is to detect dynamic phenomena in or within biological samples with spatial extension. In cell cultures of muscle cells, such movements can be triggered, for example, by the contraction of individual muscle cells.

[0013] The biological sample can also be a sample of free-swimming microorganisms, such as sperm. In this case, the method can be used to detect the movement of the free-swimming microorganisms, for example, in the case of sperm, to determine sperm motility.

[0014] The biological sample will hereinafter also be referred to simply as the sample. Preferably, the diameter of the sample is at most 5 mm, more preferably at most 1 mm.

[0015] The method according to the invention comprises providing an optical wide-field illumination device for illuminating the sample, which is configured to illuminate the entire sample, and furthermore providing a detector for detecting the radiation emanating from the sample.

[0016] Here, the detector has a detection area that is subdivided into several detection regions. For example, the detector can be a two-dimensional pixel array detector with matrix-like arranged pixel elements, e.g., an optical pixel sensor. According to a variant of the invention, the detection area can be subdivided into at least 10 x 10 or at least 100 detection regions. The detector's detection area can be subdivided into, for example, 10,000 or more detection regions, e.g., pixels, arranged in multiple rows side by side, e.g., into 100 x 100 pixels or channels or more. The higher the spatial resolution or the number of pixels, the more reliably it can be ensured that motion-induced changes in the detected intensity do not average out but are detected cumulatively.

[0017] The device, in particular the detector, is configured to derive detection signals from individual detection areas with respect to time, then rectify them, preferably by taking the magnitude or squaring the signals, and summ or average the derived and rectified detection signals from all detection areas. Deriving the detection signals from individual detection areas with respect to time means that the nth derivative, n ≥ 1, with respect to the time of the detection signal of each detection area, e.g., each pixel, is determined. Preferably, the first time derivative is determined in each case, i.e., n = 1.

[0018] According to the invention, a detector with a spatially resolved detection area in the form of individual detection regions, hereinafter also referred to as channels or pixels, is used. The measurement signals or curves of the individual detection regions can be in digital or analog form and are differentiated with respect to time in a first step and then rectified. Subsequently, the rectified signals or data of the individual detection regions are summed or averaged, thus yielding a quantitative measure of the dynamics in a pattern or image generated by the sample on the detection area. Externally, the area-cumulative motion sensor thus appears as a detector that outputs a signal or data without spatial resolution. The detector can therefore be designed to output a single-channel signal.Complex analyses such as pattern recognition and trajectory formation for measuring and describing dynamic parameters are no longer necessary.

[0019] The detector is therefore preferably a non-imaging detector. The detector's output signal can be analog or digital.

[0020] A further advantage is that dynamic phenomena in the sample are detected by changes in brightness or intensity on the detection surface. This is achieved by using the measurement approach according to the invention to measure the sum or average of the magnitude of the brightness changes generated by dynamic phenomena in the sample at constant illuminance, cumulatively across the entire detection surface. This prevents positive or negative individual measurements from canceling each other out. Furthermore, the sensitivity of the measurement is increased.

[0021] An optical widefield illumination system is understood to be a non-scanning illumination system, i.e., an illumination system that can illuminate the entire sample without the use of optics for focusing on individual image planes within the sample or for successively scanning the sample volume. With such a widefield illumination system, the detector simultaneously captures all signal contributions from the entire sample volume.

[0022] A further advantage of the method according to the invention is therefore that the method can be carried out using comparatively simple optical elements and that optics for successive scanning of the sample volume can be dispensed with. Therefore, the device for carrying out the method can be designed to be cost-effective and compact. Likewise, the method exhibits only low sensitivity to misalignment.

[0023] Due to the ease of evaluation and high sensitivity of the measurement signal, as well as the compact design, the method is particularly suitable for the parallel monitoring of a large number of samples and can be integrated efficiently into screening environments or automated high-throughput processes, e.g., high-throughput screening methods.

[0024] According to a preferred embodiment, the time derivative and subsequent rectification of the detection signals from individual detection areas are performed using an integrated circuit. This integrated circuit can, for example, be a CMOS circuit (complementary metal-oxide-semiconductor circuit). This enables fast processing of the measurement signals and a cost-effective and compact detector design.

[0025] Movement within the sample and / or movement of a sample component can be detected, for example, if a value of the detector's output signal exceeds a predetermined threshold. In the case of muscle cells, movement within the sample can also be detected if the output signal exhibits periodicity.

[0026] According to a particularly preferred embodiment, movement in the sample is detected by means of a change in the diffraction pattern. According to this embodiment, the optical wide-field illumination device comprises a light source that generates coherent light, wherein at least part of a diffraction pattern generated by light diffracted through the sample from the light source falls onto the detection surface.

[0027] In a diffraction pattern, each point of the pattern contains signal contributions from the entire sample. Therefore, it is generally sufficient to detect only a portion of the diffraction pattern generated by the sample. Movement within the sample causes a change in the diffraction pattern, for example, a speckle pattern, but the overall brightness of the changing diffraction pattern remains approximately constant over time. The measurement approach according to the invention, which involves dividing the detection area into smaller detection regions and aligning the temporal changes of the measurement curves before adding or averaging them, prevents the changes caused by movement within the sample from averaging out.

[0028] A particularly advantageous variant involves mapping the entire diffraction pattern onto the detection surface. This increases the sensitivity of the motion detection. Furthermore, using the entire pattern leads to drastically improved reproducibility of the measurements (compared to measuring a small section of the pattern) and a degree of quantitative comparability between similar samples.

[0029] In addition to the exemplary configuration highlighted above, in which movement in the sample is detected by a change in the generated diffraction pattern, the wide-field illumination device can also be configured to project an image of the sample onto the detection surface. According to this alternative configuration, the optical wide-field illumination device comprises a light source capable of generating coherent or incoherent light and optics configured to create an imaging beam path to project the sample onto the detector's detection surface.

[0030] Various wide-field illumination and imaging approaches can be used here. For example, the optical wide-field illumination device can be a transmitted light microscope, a dark-field microscope, or a wide-field fluorescence microscope. Depending on the type of wide-field illumination device and the positioning of the detector relative to the sample, the radiation detected by the detector and emanating from the sample can be radiation, particularly light, from a radiation source of the wide-field illumination device, which is altered in its beam direction and / or polarization state at any point within the sample by an interaction with the sample. It can also be transmitted light or fluorescence radiation.

[0031] The wide-field illumination device can include illumination optics arranged on the illumination side, by means of which the radiation from the light source is directed onto the entire sample to illuminate it completely and as uniformly as possible. The optics can further include detection optics, by means of which the light emitted by the sample, which is altered in its beam direction, polarization state, and / or diffraction pattern by interaction with the sample, is directed onto a detection surface of the detector. These functional properties of the illumination optics and / or the detection optics can be realized using one or more suitably arranged and designed known optical elements and components, such as filters, lenses, apertures, refractive elements, etc.

[0032] The method can further include providing a receptacle for the sample. The receptacle for holding the sample can comprise a support matrix, preferably a biopolymer. Alternatively, the sample receptacle can be configured as a support matrix, preferably a biopolymer such as alginate, contained within a hanging droplet. For use of the method in screening environments, the sample receptacle can be a cavity of a multiwell plate (microtiter plate). Furthermore, the receptacle can be a cavity of a multiwell plate designed to form a hanging droplet at each cavity (so-called hanging-drop multiwell plate). Such hanging-drop multiwell plates are offered, for example, by Insphero AG, CH-8952 Schlieren, under the name "GravityPLUS™ 3D Culture and Assay Platform". Patent EP 2 342 317 B1 also discloses such a plate.

[0033] As mentioned above, the method is particularly suitable for the parallel monitoring of a large number of samples, e.g., samples to be analyzed using automated high-throughput methods. An advantageous further development of the method therefore provides for the parallel optical detection of movements in several separate biological samples or of movements of sample components in several separate biological samples. According to this variant, the sample receptacle can be a multiwell plate or a hanging-drop multiwell plate, which has a plurality of cavities arranged in rows and columns for receiving the samples. Furthermore, several detectors, as described in this document, are provided in the form of a detector array, with each detector in the array being assigned to a cavity.In this case, the grid spacing of the individual detectors preferably corresponds to the grid spacing of the cavities of the multiwell plate.

[0034] The diameter of the detector's detection area can be less than or equal to 9 mm. This is particularly advantageous if the detector is used for motion detection of samples stored in multi-well plates, especially samples stored in 96-well plates.

[0035] In the parallel optical detection of motion in several separate biological samples, each stored in a cavity or hanging droplet of a multiwell plate, the optical wide-field illumination device can be configured to illuminate the individual cavities. If coherent light is required, for example, for the embodiment in which a diffraction pattern is detected, the optical wide-field illumination device can be implemented as a laser diode array, with the grid spacing of the individual laser diodes corresponding to the grid spacing of the cavities of the multiwell plate. A laser diode array represents a space-saving and energy-efficient illumination source.

[0036] Instead of a laser diode array as the light source, the light source can be a conventional light source (laser, arc lamp, etc.), with the light from the light source being coupled into the individual cavities containing the samples via a fiber optic bundle. Each fiber optic strand is assigned to one cavity. This offers the advantage that the light source can be operated at a sufficient distance from the sample to avoid excessive heat generation near the sample.

[0037] Furthermore, according to another alternative approach, the multiwell plate can be illuminated across its entire surface with a light source. This is a simple implementation option, but it has disadvantages in terms of energy efficiency, as the light source must be correspondingly powerful. It is advantageous here if the light is parallelized and directed precisely onto the cavities using appropriately designed condenser optics and, if necessary, angle-dependent transmission filters.

[0038] According to a second aspect of the invention, a device is provided for the contactless in-vitro detection of movement in a biological sample and / or movement of a sample component of the biological sample.

[0039] The diameter of the biological sample can, for example, be at least 50 µm. The device comprises a wide-field optical illumination unit for illuminating the sample, configured to illuminate the entire sample. The device further comprises a detector for detecting radiation emanating from the sample, the detector having a detection area divided into several detection zones. The detector is configured to derive detection signals from individual detection zones over time, then rectify them, preferably by calculating the magnitude or squaring, and summ or average the derived and rectified detection signals of all detection zones and then provide them as an output signal.

[0040] According to a preferred embodiment of the invention, the detector has an integrated circuit, preferably a CMOS circuit, configured to perform the time derivative and subsequent rectification of the detection signals of individual detection areas.

[0041] The detector can output a single-channel signal and / or a signal that is not spatially resolved with respect to the detection area.

[0042] According to a third aspect of the invention, a detector for detecting optical radiation is provided, with a detection area which is divided into several detection areas (4a), wherein the detector is configured to derive detection signals of individual detection areas with respect to time, then rectify them, preferably by taking the magnitude or squaring them, and summe or average the derived and rectified detection signals of all detection areas and then provide them as an output signal.

[0043] To avoid repetition, features disclosed solely in relation to the device shall also be deemed disclosed and claimable in relation to the method, and vice versa. The aforementioned aspects and features of the invention, particularly with regard to the design of the detector or the wide-field optical illumination device, which were described in connection with the method, thus also apply, for example, to the device.

[0044] The preferred embodiments and features of the invention described above can be combined in any way desired. Further details and advantages of the invention are described below with reference to the accompanying drawings. These show: Figure 1 is a schematic representation of a method and a device according to one embodiment of the invention; Figure 2 is a representation of a speckle pattern mapped onto the detector surface; Figure 3 is a schematic illustration of the data processing of the detector according to one embodiment of the invention; Figure 4 shows the effect of differentiating and rectifying the measured values ​​of individual pixels; and Figure 5 is a schematic representation of a method and a device according to a further embodiment of the invention.

[0045] Identical parts are marked with the same reference symbols in the figures and are not described separately.

[0046] Figure 1 shows a schematic representation of a method and a device according to an embodiment of the invention.

[0047] To carry out the procedure, a device for the optical in vitro detection of movement in a biological sample 1 is provided. The biological sample 1 can be a cluster of cells from muscle tissue.

[0048] The device comprises a receptacle (not shown) for the sample 1, an optical wide-field illumination device 2 and a detector 3.

[0049] The uptake is not limited to a specific type of uptake, but can be appropriately designed depending on the application and type of sample, e.g. as a support, support plate, vessel, cavity of a multiwell plate or as a support matrix in the form of a biopolymer, e.g. alginate, on which the sample is grown.

[0050] At the in Figure 1In the illustrated embodiment, the optical wide-field illumination device is a radiation source that generates coherent light (laser). The entire sample 1 is illuminated by means of the coherent radiation 8 generated by the radiation source 2. For this purpose, it may be advantageous to provide an illumination optic (not shown) by means of which the radiation from the light source 2 is directed onto the entire sample 1 in order to illuminate the sample completely and as uniformly as possible. This functional property of the optic can be realized using one or more suitably arranged and designed known optical elements and components, such as filters, lenses, apertures, refractive elements, etc.

[0051] The light from light source 2, diffracted at sample 1, produces a diffraction pattern with a center of high intensity and a border region of low intensity, e.g. in the form of a speckle pattern 20, which is exemplified in Figure 2 shown.

[0052] The device further comprises an optical detector 3 for detecting the diffraction pattern 20. The detector 3 is arranged such that the diffraction pattern 20, which is generated by light 9 of the light source 2 diffracted by the sample 1, is imaged onto the detection surface 3a of the detector 3.

[0053] Here, the detection area is designed as a two-dimensional pixel array, such that the detection area 3a is subdivided into several detection areas 4a by the individual pixels 4. The detection area is subdivided into several pixels 4, e.g., into 10x10 pixels 4 or more, which is shown in the schematic representation of the Figure 1not shown. Each pixel therefore measures a section of the speckle pattern 20.

[0054] The data processing based on the measured light intensity of the individual pixels 4 is also described below with reference to Figure 3 described.

[0055] Figure 3 Figure 1 shows a perspective view of the detection area 3a, which is subdivided into individual pixels 4a. Each pixel 4 outputs the time course of the light intensity measured on the respective pixel area (detection area) 4a in the form of a detection signal 4c. Reference symbol 30a merely illustrates such a time course for a first pixel, which measures a decrease in light intensity, as represented by the downward-sloping curve. Simultaneously, a second pixel can measure an increase in light intensity, which is represented by reference symbol 30b.

[0056] Such a change over time results from a movement within the sample or from a movement of a sample component, which changes the diffraction pattern 20 produced by the sample.

[0057] In a first step S1, the measured detection signals 4c of the individual pixels 4 are differentiated with respect to time. The result of this differentiation operation is illustrated by curves 31a and 31b. Subsequently, in step S2, the individual results are rectified; that is, the differentiated measurement result of each pixel 4 is rectified, for example, by taking an absolute value or squaring it. The result of this rectification is illustrated by curves 32a and 32b.

[0058] In a third step S3, the derived and rectified detection signals of all pixels 4 are summed (or alternatively averaged) and then provided as the output signal 6c of detector 3. The result of this summation is illustrated by curve 33.

[0059] The temporal differentiation and subsequent rectification of the measurement data of the individual pixels is performed by an integrated CMOS circuit 5 of the detector 3. This CMOS circuit thus outputs a differentiated and rectified signal 5c for each pixel. These output data from the CMOS circuit are subsequently summed in a summing amplifier 6.

[0060] Detector 3 thus appears externally as a detector with only one data output 6c, through which a spatially unresolved signal / data 7 is output. The signal, which is not spatially resolved with respect to the detector area 3a, measures area-cumulatively, i.e., over the entire detection area, the sum (or average) of magnitude changes in brightness that are generated by dynamic phenomena in the sample at constant illumination intensity.

[0061] The movements in the sample then lead to the peaks in the output signal 7, which can be easily detected.

[0062] To illustrate the advantages of the approach according to the invention, Figure 4The effect of differentiating and aligning the measured values ​​of randomly selected image areas (simplified as "pixels") 4 of the detection area 3a, here pixels 48, 72, 96, 120, 144. To demonstrate the method, a video of a speckle pattern recorded on a cardiac muscle tissue model was divided into 192 image segments ("pixels") using video software, and the average brightness of each segment was recorded as a function of time (Ji(t)). Using data analysis software, these Ji(t) were derived and squared, and then these processed data sets were summed (Σ(∂Ji / ∂t)2).

[0063] The five diagrams at the top of the left column of the Figure 4 Figure 30 shows the temporal profile of the measured light intensity, measured by pixels 48, 72, 96, 120, and 144 of detector 3. Adding the measured values ​​of all image segments results in the graph shown in the lower left of the diagram. Figure 4The temporal progression shown is not reliably detectable.

[0064] The five diagrams at the top of the right-hand column of the Figure 4 show the differentiated and subsequently aligned temporal progression 32 of the measured light intensity of the individual pixels 48, 72, 96, 120, 144.

[0065] Adding up the processed values ​​of all image segments results in the value shown in the lower right diagram. Figure 4 The temporal progression shown is 33. The movement in the sample can now be clearly detected based on the individual peaks of the progression 33.

[0066] Figure 5 Shows a schematic representation of a method and a device according to a further embodiment of the invention. Here, components with the same reference numerals correspond to the components of the Figure 1 and are not described separately.

[0067] The entire biological sample 1 is illuminated by means of a wide-field optical illumination device, i.e., without the use of optics for focusing onto individual image planes within the sample or for successively scanning the sample volume. A special feature of this embodiment is that the radiation source of the wide-field optical illumination device does not produce coherent light. Consequently, no detectable diffraction pattern is generated by the sample 1. Instead, an optic 10, e.g., in the form of a convex lens, is provided for the wide-field optical illumination device, by means of which an imaging beam path 9a, 9b is generated to image one focal plane of the sample 1 onto the detection surface 3a of the detector 3. The wide-field optical illumination device can, for example, be a transmitted light microscope, a dark-field microscope, or a wide-field fluorescence microscope.The processing of the measurement data is analogous to the embodiment described above. The version with imaging optics is therefore also suitable for monitoring three-dimensional samples; however, the version based on diffraction patterns is considered more advantageous.

[0068] Although the invention has been described with reference to specific embodiments, it is apparent to a person skilled in the art that various modifications can be made and equivalents used as substitutes without departing from the scope of the invention. Furthermore, many modifications can be made without departing from the relevant scope. Consequently, the invention is not intended to be limited to the disclosed embodiments but is intended to encompass all embodiments falling within the scope of the appended claims. In particular, the invention also claims protection for the subject matter and features of the dependent claims independently of the referenced claims.

[0069] The disclosure of the invention further comprises the following aspects: According to a first aspect, a method for the optical in vitro detection of movement in a biological sample (1) and / or movement of a sample component of the biological sample (1), comprising the steps: a) Providing a1) an optical wide-field illumination device for illuminating the sample, configured to illuminate the entire sample (1), and a2) a detector (3) for detecting radiation (9; 9a, 9b) emanating from the biological sample, wherein the detector 3) has a detection area (3a) divided into several detection areas (4a) and configured to derive detection signals (4c) of individual detection areas (4a) with respect to time (S1), subsequently rectify them (S2), preferably by taking a magnitude or squaring them, and summ or average (S3) the derived and rectified detection signals of all detection areas and then provide them as an output signal (6c). b) Illuminating the biological sample (1) with the wide-field illumination device; and c) detecting motion in the biological sample (1) as a function of the output signal (6c) of the detector (3).

[0070] An optional aspect of the method is that the time derivative (S1) and subsequent rectification (S2) of the detection signals of individual detection areas can be carried out in the detector (3) using an integrated circuit (5), preferably a CMOS circuit.

[0071] Further optional aspects of the procedure include: a) that the detector (3) outputs a single-channel signal (33); and / or b) that the detector outputs a signal (33) or data that is not spatially resolved with respect to the detection area (3a); and / or c) that the detector (3) is a non-imaging detector.

[0072] In an optional aspect of the method, the optical wide-field illumination device may comprise a light source (2) that generates coherent light, wherein at least part of a diffraction pattern (20) generated by light (9) diffracted by the sample (1) from the light source (2) is imaged onto the detection surface (3a) of the detector (3).

[0073] In an alternative optional aspect of the method, the optical wide-field illumination device may comprise a light source and optics (10), wherein the optics (10) is configured to generate an imaging beam path (9a, 9b) to image the sample (1) onto the detection surface (3a) of the detector (3).

[0074] The optical wide-field illumination device can be a transmitted light microscope, a dark-field microscope, or a wide-field fluorescence microscope.

[0075] An optional aspect of the procedure is that movement in the sample (1) and / or movement of a sample component of the sample (1) can be detected if a value of the output signal (33) of the detector exceeds a predetermined threshold and / or if the output signal (33) exhibits periodicity.

[0076] Further optional aspects of the procedure provide that the biological sample (1) a) is a three-dimensional cell and / or tissue culture or cell cluster; or b) is a sample of free-floating microorganisms, for example sperm.

[0077] Further optional aspects of the method provide that a receptacle for holding the sample (1) comprises a) a support matrix, preferably a biopolymer (15); and / or b) a hanging drop; and / or c) a cavity of a multiwell plate or a hanging-drop multiwell plate.

[0078] Another embodiment provides a method in which parallel optical detection of a movement in several separate biological samples or of a movement of a sample component in several separate biological samples is carried out, wherein a receptacle for the samples is a multiwell plate or a hanging-drop multiwell plate having a plurality of cavities arranged in rows and columns for receiving the samples, and wherein several detectors (3) are provided in the form of a detector array, with each detector of the detector array being assigned to a cavity.

[0079] In an optional aspect of the procedure, the diameter of the detection area (3a) of the detector (3) can be less than or equal to 9 mm.

[0080] Further optional aspects of the procedure provide that a) the biological sample (1) comprises a plurality of cells; and / or b) the diameter of the biological samples is at least 50 micrometers in all spatial directions; and / or c) the diameter of the biological sample is larger than a diffraction-limited resolution achievable by the wide-field optical illumination device.

[0081] According to a second aspect, the disclosure includes a device for the contactless in vitro detection of motion in a biological sample (1) and / or motion of a sample component of the biological sample (1). The device comprises a wide-field optical illumination device for illuminating the sample (1), configured to illuminate the entire sample (1), and a detector (3) for detecting radiation (9; 9a, 9b) emanating from the sample (1), wherein the detector (3) has a detection area (3a) which is subdivided into several detection areas (4a) and is configured (5, 6) to derive detection signals (4c) of individual detection areas (4a) over time, subsequently rectify them, preferably by taking a magnitude or squaring them, and summ or average the derived and rectified detection signals of all detection areas and then provide them as an output signal (33).

[0082] In one embodiment of the device, the detector may have an integrated circuit (5), preferably a CMOS circuit, configured to perform the time derivative and subsequent rectification of the detection signals (4c) of individual detection areas (4a).

[0083] Further optional aspects of the device include: a) that the detector (3) outputs a single-channel signal (33); and / or b) that the detector (3) outputs a signal (33) that is not spatially resolved with respect to the detection area; and / or c) that the detection area (3a) is divided into at least 100 or 10*10 pixels (4) arranged in multiple rows next to each other.

[0084] According to a third aspect, the disclosure comprises a detector (3) for detecting optical radiation, with a detection area (3a) which is divided into several detection areas (4a), wherein the detector (3) is configured (5, 6) to derive detection signals (4c) of individual detection areas (4c) with respect to time, subsequently rectify them, preferably by taking the magnitude or squaring them, and summe or average the derived and rectified detection signals of all detection areas and then provide them as an output signal (33).

Claims

1. Detector (3) for detecting optical radiation with a detection area (3a) which is subdivided into a plurality of detection regions (4a), wherein the detector (3) is configured (5, 6) to form the derivative with respect to time of detection signals (4c) of individual detection regions (4c), subsequently to rectify them, preferably by absolute value generation or squaring, and to sum or to average the differentiated and rectified detection signals of all the detection regions and then to provide them as an output signal (33); and wherein the detector (3) is a non-image forming detector.

2. Detector (3) according to claim 1, characterised in that the detector (3) comprises an integrated circuit (5), preferably a CMOS circuit, that is configured to perform the derivative with respect to time and the subsequent rectification of the detection signals (4c) of individual detection regions (4a).

3. Detector (3) according to one of the preceding claims, characterised in that the detector (3) is configured to output a single-channel signal (33).

4. Detector (3) according to one of the preceding claims, characterised in that the detector (3) is configured to output a signal (33) that is not spatially resolved or data that is not spatially resolved with respect to the detection area (3a).

5. Detector (3) according to one of the preceding claims, characterised in that a diameter of the detection area (3a) of the detector (3) is smaller than or equal to 9 mm.

6. Detector (3) according to one of the preceding claims, characterised in that the detection area (3a) is subdivided into at least 100 or 10 x 10 multi-line adjacently arranged pixels (4).

7. Detector (3) according to one of the preceding claims, characterised in that the detection area (3a) is subdivided into at least 10000 or 100*100 multi-line adjacently arranged pixels (4).