Method and system for detecting cells in a biopsy sample

Abstract

The invention relates to a method and system for detecting cells of a first cell type (1) in a biopsy sample in which a plurality of cells of different cell types (2, 3) are present. The invention further relates to a device for carrying out a magnetic field measurement on cells of a biopsy sample that are labeled with magnetic nanoparticles. The method comprises the following method steps: carrying out tissue dissociation of the biopsy sample in order to singulate the cells present in the biopsy sample; supplying magnetic nanoparticles (11) to the singulated cells in order to selectively label the cells of the first cell type (1); removing the excess magnetic nanoparticles (11); introducing the cells into a microchannel (5), the microchannel (5) having a cross section which allows only one labeled cell (1) to pass through at a time; carrying out a magnetic field measurement in the microchannel (5) by means of a quantum magnetic field sensor (6) based on the F-center principle; and sorting out the labeled cells (1) after they have passed through the microchannel (5).

Description

[0001] DELETE

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[0005] Method and system for detecting cells in a biopsy sample

[0006] DESCRIPTION

[0007] 5

[0008] The invention relates to a method and system for detecting cells of a first cell type in a biopsy sample containing a plurality of cells of different cell types. Furthermore, the invention relates to a device for performing a magnetic field measurement on cells of a biopsy sample labeled with magnetic nanoparticles.

[0009] A common task in medical technology is to identify altered, abnormal, or pathological cells in a tissue sample taken from a living organism. Histological examination of biopsy samples is a well-known method for this purpose. However, in the context of tumor diseases, for example, sensitive, rapid, and reliable examinations are needed to provide the most accurate and quickest possible diagnosis. One challenge here is that a biopsy sample (e.g., during an initial diagnosis of various tumor diseases) may contain very few tumor cells. These cells must be identified with a high degree of reliability. At the same time, the examination results should generally be available as quickly as possible to allow for immediate medical intervention based on the examination findings.Tissue samples are often requested during surgery when the surgeon needs short-term information to determine the further course and extent of the procedure. A common example is checking whether lymph nodes are affected by a tumor or whether a tumor has been completely removed. DELETE.

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[0014] The invention is based on the objective of providing a sensitive, reliable, and at the same time rapid method and system for the detection of cells of a first cell type in a biopsy sample containing a plurality of cells of different cell types. Furthermore, it is

[0015] 5. The object of the invention is to provide a device that allows for sensitive, reliable, and rapid magnetic field measurements on cells of a biopsy sample labeled with magnetic nanoparticles, in order to detect cells of a first cell type in a biopsy sample. 0. This object is achieved by a method with the features of claim 1, by a device with the features of claim 11, and by a system with the features of claim 15. Advantageous embodiments are described in the dependent claims.

[0016] The inventive method for detecting cells of a first cell type in a biopsy sample containing a plurality of cells of different cell types comprises the following process steps: performing tissue dissociation of the biopsy sample to isolate the cells contained in the biopsy sample; introducing magnetic nanoparticles to the isolated cells to selectively label the cells of the first cell type; removing the excess magnetic nanoparticles; introducing the cells into a microchannel, wherein the microchannel has a cross-section that allows the simultaneous passage of only one labeled cell; performing a magnetic field measurement in the microchannel using a quantum magnetic field sensor based on the color center principle; and sorting out the labeled cells after passing through the microchannel.

[0017] In this context, cells of a cell type are understood to be cells that differ from cells of other cell types by one property. For example, tumor cells of a first tumor cell line can have a first DELETE

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[0021] Forming a cell type, tumor cells of a second tumor cell line form a second cell type, and non-cancer tissue cells form a third cell type.

[0022] Furthermore, it is also possible to detect certain viruses (instead of cells).

[0023] 5. In this process, for example, viruses (e.g., coronaviruses) of a first type can form a first virus type, viruses of a second type a second virus type, which are magnetically labeled as described for cells and then detected analogously according to the invention.

[0024] The subject matter of the invention described in the claims relates to cells of all kinds and can alternatively also relate to viruses of all kinds.

[0025] Furthermore, the detection of different cell types is not limited to the field of oncology. Another application could be the detection of biomarkers indicative of neurodegenerative diseases. For this purpose, SPIONs can be functionalized with antibodies correlated with such diseases, such as anti-TAU antibodies, anti-amyloid-beta antibodies, or anti-Lewybody antibodies. By coupling defined surface markers of the cells of the first cell type with magnetic nanoparticles, the magnetic signature of these cells (e.g., tumor cells) can be read out with high efficiency and specificity using quantum magnetometry. Quantum sensing enables the determination of parameters such as magnetic field strength in various physical, biological, or material systems. It aims, in particular, to optimize the precision and selectivity of measurements through quantum effects.Quantum magnetic field sensors based on the color center principle (e.g., the NV center in a diamond) have the following advantageous properties: high magnetic sensitivity, operation at room temperature, large dynamic range (several Tesla), and zero calibration-free operation. DELETE.

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[0030] Performing the magnetic field measurement in the microchannel enables cell-accurate detection, which in turn leads to high selectivity and sensitivity of the measurement.

[0031] 5 In other words, a quantum magnetic field sensor is used to selectively identify cells of a specific cell type (e.g., cells of individual cancer cell lines). This means that the selective binding of magnetic nanoparticles to cells and the resulting magnetic field detect which cells are labeled and which are not. Since only cells belonging to a specific cell type are labeled, these cells can be selectively identified via the magnetic field and the sorting process after they have passed through the microchannel.

[0032] The method according to the invention thus enables sensitive, reliable, and rapid detection of cells of a first cell type in a biopsy sample. For example, the method according to the invention allows an ongoing operation to be interrupted for histological examination for only a few minutes, whereas previously the operation had to be interrupted for a longer period, often for several hours. The biopsy sample can be a tissue sample removed under anesthesia (tumor margin determination).

[0033] The biopsy sample can also advantageously be a fine-needle aspiration sample. In fine-needle aspiration, a fine needle is inserted into abnormal tissue or suspicious body fluids. Fine-needle aspiration is considered minimally invasive, but also yields less sample material than conventional biopsy samples. Due to the high sensitivity of the method according to the invention, only a small sample volume is required. Accordingly, a fine-needle aspiration sample is very well suited here. DELETE

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[0038] The magnetic nanoparticles are advantageously designed as superparamagnetic iron oxide nanoparticles (SPIONs). These offer advantages such as small particle size and magnetic controllability.

[0039] 5. Biocompatibility, the possibility of diverse surface modifications and colloidal stability.

[0040] In an advantageous embodiment, the method can further include a process step for changing the cross-section of the microchannel depending on the cell size. This allows the application range to be increased, as it ensures that only one cell can enter the microchannel at a time for different cell sizes.

[0041] A flow regulator can be installed at the microchannel's inlet. This ensures that only a limited number of cells enter the microchannel, allowing for more reliable magnetic field measurements. A microfluidic switch can be installed at the microchannel's outlet, enabling the immediate removal of the marked cells.

[0042] In other words, the magnetically identified cells are separated from the remaining cells at the microchannel's output by sorting. For this purpose, a flow controller is connected to the microchannel's input and a microfluidic switch for cell sorting is connected to the output. Based on the cell's position during the magnetic field measurement and the calibrated cell flow after the measurement, the position of the microfluidic switch for cell sorting can be determined. DELETE

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[0047] The cross-section of the microchannel is dimensioned such that it allows the simultaneous passage of only one marked cell, with the cross-section being round / oval, square, or rectangular in advantageous embodiments. However, embodiments with a semicircular cross-section or similar shapes are also possible.

[0048] In an advantageous embodiment, the quantum magnetic field sensor has a sensor layer in which color centers are formed and which constitutes at least a portion of an inner surface of the microchannel. In this way, the sensor layer can be arranged in close proximity to the cells to be detected. Consequently, the quantum magnetic field sensor requires very little installation space.

[0049] For magnetic field measurement using the quantum magnetic field sensor, a so-called continuous wave measurement can be performed.

[0050] This type of measurement has a relatively simple setup and delivers particularly fast measurement results.

[0051] It is also possible to perform the magnetic field measurement using the quantum magnetic field sensor via a so-called pulsed measurement. This type of measurement is 10 to 100 times more sensitive than the continuous measurement mentioned above. However, it has a more complex setup.

[0052] After sorting out the labeled cells, a further step can be taken: a light microscopic examination of the sorted, labeled cells to investigate, for example, further properties of the sorted cells. DELETE

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[0056] Furthermore, after sorting out the marked cells, molecular follow-up investigations of all kinds can be carried out, e.g. a determination of the mutation status of the cells and their gene signatures.

[0057] When examining tumor cells from blood, circulating [cells / pathogens] could also be detected.

[0058] 5 tumor cells were sorted out and thus isolated.

[0059] Further advantageous embodiments can aim to sort out not only one cell type, but at least one other cell type. For example, in the process step "introducing magnetic nanoparticles," first magnetic nanoparticles can be introduced to selectively label cells of the first cell type, and second magnetic nanoparticles can be introduced to selectively label cells of a second cell type. The first magnetic nanoparticles differ from the second nanoparticles both in their surface properties (to enable selective labeling of cells of the first cell type or selective labeling of cells of the second cell type) and in their magnetic properties. During the magnetic field measurement in the microchannel, it is then possible to distinguish between the labeled cells of the first cell type and the labeled cells of the second cell type.As a result, a simultaneous, selective sorting of the first and second cell types can take place.

[0060] The process steps "adding magnetic nanoparticles" through "adding the labeled cells" can also be repeated on the cells remaining after the "sorting out the labeled cells" step. In the "adding magnetic nanoparticles" step, further magnetic nanoparticles are then added for the selective labeling of cells of another cell type. This allows for the serial, selective sorting out of multiple cell types. DELETE

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[0065] The device according to the invention for carrying out a magnetic field measurement on cells of a biopsy sample labeled with magnetic nanoparticles has, in its most general form, a microchannel which

[0066] 5 has a cross-section that allows the simultaneous passage of only one labeled cell, and a quantum magnetic field sensor based on the color center principle for performing a magnetic field measurement in the microchannel. This device is particularly suitable by carrying out the process steps "inserting the cells into a microchannel" and "performing a magnetic field measurement in the microchannel".

[0067] The system according to the invention for detecting cells of a first cell type in a biopsy sample containing a plurality of cells of different cell types comprises: a tissue dissociation device for performing tissue dissociation of the biopsy sample to isolate the cells contained in the biopsy sample; a labeling device for supplying magnetic nanoparticles to the isolated cells and for selectively labeling the cells of the first cell type; a removal device for removing the excess magnetic nanoparticles; the aforementioned device; and a sorting device for sorting out the labeled cells after they have passed through the microchannel.

[0068] The invention is further explained with reference to exemplary embodiments in the drawings, where identical reference numerals denote identical or equivalently acting components. The drawings show:

[0069] Fig. 1 is a block diagram of the first process flow according to the invention; DELETE

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[0074] Fig. 2 a block diagram of a second invention

[0075] Procedure;

[0076] Fig. 3 a block diagram of a third invention

[0077] 5. Procedure;

[0078] Fig. 4 shows a schematic sectional view of a device for

[0079] Performing a magnetic field measurement and using a sorting device; 0

[0080] Figs. 5a to 5i Schematic representations of a sorting of cells from a biopsy sample; and

[0081] Figs. 6a to 6f Schematic sectional views of a microchannel with quantum magnetic field sensor.

[0082] Fig. 1 illustrates a first process sequence according to the invention in the form of a block or flowchart. The starting point is a biopsy sample, which was taken, for example, during a medical examination. This is in particular a fine-needle biopsy sample. However, it can also be a core needle biopsy sample.

[0083] The biopsy sample undergoes tissue dissociation to isolate the cells contained within it. Tissue dissociation methods are well-known, often employing manual techniques. However, automated methods are more advantageous, enabling more effective and reproducible tissue sample preparation. For example, automated methods combining mechanical processes with enzymatic digestion are also known. DELETE

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[0087] Once the cells from the biopsy sample are separated into individual cells, magnetic nanoparticles are introduced. These magnetic nanoparticles are called SPIONs, which

[0088] 5 are tailored to the cell type to be detected, so that selective labeling of the cells of this cell type (hereinafter also referred to as first cell type or target cells) takes place.

[0089] A prerequisite for the selective labeling of target cells using SPIONs is a suitable surface property of the magnetic nanoparticles. First, this property must be designed so that the nanoparticles are colloidally stable within the matrix. Otherwise, due to Oswald ripening, the particles will grow and eventually sediment. Uncoated nanoparticles are also susceptible to non-specific adsorption processes, resulting in all cells being magnetically labeled. Therefore, SPIONs for this application must primarily be colloidally stable and as binding-neutral as possible. For specific binding to the target cells, the expression of corresponding cell receptors must first be known. These receptors should either be present exclusively on the cells of the first cell type or be significantly overexpressed compared to the cells of the other cell types.Then, appropriate ligands (i.e., antibodies against these receptors) are selected and bound to the surface of the SPIONs. This can be achieved either non-specifically, for example, via an active ester binding strategy to target the always present amino groups in the antibody protein, or via chemoselective tagging, if, for example, specific functional targets have been previously implemented in this protein. An example of the latter is the complementary biotin-streptavidin binding pair. However, there are numerous other orthogonal (i.e., directed) binding strategies based on different tags. Examples include Snap-Tag, Spy-Tag, etc. DELETE.

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[0094] Superfluous nanoparticles – that is, nanoparticles that have not bound to the cells of the first cell type – are removed in a subsequent purification or washing step. This leaves behind isolated

[0095] 5 cells, wherein the cells of the first cell type are labeled with magnetic nanoparticles, while the cells of the remaining cell types are not labeled with magnetic nanoparticles.

[0096] This mixture of cells from different cell types is then fed into a device for performing a magnetic field measurement on cells of a biopsy sample labeled with magnetic nanoparticles. This device is shown in a schematic sectional view in Fig. 4.

[0097] Fig. 4 shows cells of a first cell type 1, a second cell type 2, and a third cell type 3, with only one of each cell type being labeled with a reference symbol in the figure. The cells of the first cell type 1 are labeled with first nanoparticles 11. For illustrative purposes, these first nanoparticles 11 are depicted as octagons. In reality, however, the first nanoparticles 11 have a different shape. Furthermore, in the figure, only one first nanoparticle 11 is associated with each cell of the first cell type 1. However, it is also possible that several first nanoparticles 11 are attached to a single first cell of the first cell type 1.

[0098] The device for performing a magnetic field measurement has a microchannel 5. The microchannel 5 has a cross-section that allows the simultaneous passage of only one of the labeled cells of the first cell type 1. Typically, the diameter of the microchannel 5 is in the range of 10 micrometers to 100 micrometers, but depends on the cell types detected and on the other cell types in the biopsy sample. DELETE

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[0102] Furthermore, the device for performing a magnetic field measurement includes a quantum magnetic field sensor 6 based on the color center principle. The quantum magnetic field sensor 6, in turn, has a sensor layer 7 in which color centers are formed. In the described

[0103] 5 In this embodiment, the NV (nitrogen vacancy) centers in the diamond are preferably with an NV center density of 0.1 ppm to 100 ppm. However, other color centers are also possible, e.g.: silicon vacancy centers (SiV), germanium vacancy centers (GeV), nickel vacancy centers (NiV), chromium vacancy centers (CrV), silver vacancy centers (AgV), tin vacancy centers or C60 color centers (fullerenes).

[0104] The sensor layer 7 forms a sub-area of ​​an inner surface of the microchannel 5. A flow controller 8 is arranged at the inlet of the microchannel 5 and a microfluidic switch 10 is positioned at the end of the microchannel 5 as part of a sorting device 9.

[0105] A magnetic field measurement is performed in the microchannel 5 of the cells that have passed through the flow controller 8 using the quantum magnetic field sensor 6. The measurement method used can be a continuous wave measurement, in particular an ODMR (Optically Detected Magnetic Resonance) measurement, or a pulsed measurement. Examples of continuous wave measurement methods, besides ODMR, include: Continuous Wave Electron Paramagnetic Resonance (CW-EPR), Continuous Wave Nuclear Magnetic Resonance (CW-NMR), Continuous Wave Electron Spin Resonance (CW-ESR), and Continuous Wave Photoacoustic Spectroscopy (CW-PAS). Examples of pulsed measurement techniques include: Pulsed Electron Paramagnetic Resonance (Pulsed EPR), Pulsed Nuclear Magnetic Resonance (Pulsed NMR), Dynamical Decoupling, Ramsey Interferometry, Spin Echo, Carr-Purcell-Meiboom-Gill (CPMG) sequence, and Rabi oscillations. DELETE

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[0110] The magnetic field measurement determines whether the cell passing the quantum magnetic field sensor 6 is a cell of the first cell type 1 labeled with a first magnetic nanoparticle 11 or a

[0111] 5 unlabeled cells (i.e., a cell of the second cell type 2 or the third cell type 3). Depending on the result of the quantum magnetic field sensor 6, the sorting device 9 sorts the cells accordingly. In other words, the sorting device 9 separates the labeled cells of the first cell type 1 from the remaining cell types 2, 3 of the biopsy sample and thus sorts out the cells of the first cell type 1. This sorting is schematically illustrated in Figure 4 by the collection of the cells of the first cell type 1 in the right container and the collection of the unlabeled cells 2, 3 in the left container.

[0112] Not shown in Figure 1 is a subsequent light microscopic examination of the sorted cells of the first cell type 1 in order to identify these cells more precisely.

[0113] The device described above for performing a magnetic field measurement can be part of a system (not shown in the figures) for detecting cells of the first cell type 1 in the biopsy sample. This system further comprises a tissue dissociation device for performing tissue dissociation of the biopsy sample, a labeling device for delivering the magnetic nanoparticles 11 to the isolated cells and for selectively labeling the cells of the first cell type 1, a removal device for removing the excess magnetic nanoparticles 11, and the sorting device 9 for sorting out the labeled cells of the first cell type 1 after they have passed through the microchannel 5.

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[0118] Figures 2 and 3 show variations of the procedure that allow the tissue sample to be divided into more than two fractions. In other words, these methods can be used to separate not only cells of a first cell type 1, but also, for example, cells of a second cell type 2.

[0119] 5

[0120] The method shown in Figure 2 differs from the method shown in Figure 1 in that, in the process step "introducing magnetic nanoparticles," first magnetic nanoparticles 11 are introduced for the selective labeling of cells of the first cell type 1, and second magnetic nanoparticles 12 are introduced for the selective labeling of cells of a second cell type 2. The first magnetic nanoparticles 11 differ from the second nanoparticles 12 in both their surface properties and their magnetic properties. During the magnetic field measurement in the microchannel 5, it is then possible to distinguish between the labeled cells of the first cell type 1 and the labeled cells of the second cell type 2. As a result, the cells of the first cell type 1 and the cells of the second cell type 2 can be simultaneously and selectively sorted out.

[0121] Light microscopic examinations are then carried out on the sorted cells of the first cell type 1 and the second cell type 2 in order to identify these cells more precisely.

[0122] Figures 5a to 5i schematically illustrate the process of sorting cells from a biopsy sample according to the method shown in Figure 2. A dissociated biopsy sample containing cells of the first cell type 1, labeled with first magnetic nanoparticles 11, cells of the second cell type 2, labeled with second magnetic nanoparticles 12, and 0 unlabeled cells of a third cell type 3, is fed into the device for performing a magnetic field measurement. The second nanoparticles 12 are shown as pentagons for illustrative purposes. DELETE

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[0127] In reality, however, the second nanoparticles 12 have a different shape. Furthermore, in the figure, only one second nanoparticle 12 is connected to a cell of the second cell type 2. However, it is also possible that several second nanoparticles 12 are attached to a single cell of the second cell type 2.

[0128] 5 are attached.

[0129] Figure 5a shows a state in which the flow regulator 8 is opened and a labeled cell of the first cell type 1 enters the microchannel 5. The cell of the first cell type 1, labeled with the first nanoparticle 11, moves along the microchannel 5 and passes the quantum magnetic field sensor 6 with its sensor layer 7 (see Figure 5b). The quantum magnetic field sensor 6 detects the magnetic field generated by the first magnetic nanoparticle 11. Based on this measurement, the sorting device 9 assigns the labeled cell of the first cell type 1 to a first fraction and controls the microfluidic switch 10 such that the labeled cell of the first cell type 1 is collected in a first container (see Figure 5c).

[0130] Figure 5b shows a state in which the flow regulator 8 is opened and an unlabeled cell of the third cell type 3 enters the microchannel 5. This unlabeled cell moves along the microchannel 5 and also passes the quantum magnetic field sensor 6 with its sensor layer 7 (see Figure 5e). The quantum magnetic field sensor 6 detects no or no significant change in the magnetic field. Based on this measurement result, the sorting device 9 assigns the cell of the third cell type 3 to another fraction (e.g., a third fraction) and controls the microfluidic switch 10 such that the cell of the third cell type 3 is collected in a third container (see Figure 5f). Figure 5g shows a state in which the flow regulator 8 is opened and a labeled cell of the second cell type 2 enters the microchannel 5. The DELETE

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[0133] 251021 -WO -16- Cell of the second cell type 2, labeled with the second nanoparticle 12, moves along the microchannel 5 and passes the quantum magnetic field sensor 6 with its sensor layer 7 (see Figure 5h). The quantum magnetic field sensor 6 detects the magnetic field generated by the second nanoparticle 12.

[0134] 5 nanoparticles 12 generated a magnetic field. Based on this measurement result, the sorting device 9 assigns the marked cell of the second cell type 2 to a second fraction and controls the microfluidic switch 10 such that the marked cell of the second cell type 2 is collected in a second container (see Figure 5i).

[0135] Figure 3 shows another method variant that allows the tissue sample to be divided into more than two fractions. The method shown in Figure 3 differs from the method shown in Figure 1 in that the cells remaining after the "sorting out the labeled cells" step are subjected to the process steps "adding magnetic nanoparticles" through "sorting out the labeled cells" again, with further magnetic nanoparticles (in particular, a second set of magnetic nanoparticles 12) being added in the "adding magnetic nanoparticles" step to selectively label cells of another cell type (namely, to label the cells of the second cell type 2). As a result, a serial, selective sorting of the cells of the first cell type 1 and the cells of the second cell type 2 can be performed.The sorted cells are then subjected to light microscopic examinations in order to identify these cells more precisely.

[0136] The cross-section of the microchannel 5 is designed such that it allows the simultaneous passage of only one labeled cell of the first cell type 1. Figures 6a to 6f show schematic cross-sectional views of the microchannel 5 with a quantum magnetic field sensor 6. The quantum magnetic field sensor 6 has a sensor layer 7 in which the DELETE

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[0141] Color centers (schematically visualized as points in the figures) are formed.

[0142] Figure 6a shows a cylindrical microchannel 5, the surface of which is

[0143] 5 the sensor layer 7 of the quantum magnetic field sensor 6 is formed. Such a microchannel 5 can be produced, for example, by drilling or etching into the base material (e.g., diamond) of the quantum magnetic field sensor 6. Figure 6b shows another embodiment of a cylindrical microchannel 5, in which only a partial area (half of the lateral surface) of the surface of the microchannel 5 is formed by the sensor layer 7 of the quantum magnetic field sensor 6.

[0144] Figure 6c shows an embodiment of a microchannel 5 with a rectangular cross-section. One side of the rectangle forms the sensor layer 7 of the quantum magnetic field sensor 6. The remaining sides of the rectangle form the base material of the quantum magnetic field sensor 6. Such a microchannel 5 can be produced, for example, by etching the base material.

[0145] Figure 6d shows an embodiment of a microchannel 5 with a rectangular cross-section, wherein the microchannel 5 was first formed on the surface of the base material of the quantum magnetic field sensor 6 (in particular by mechanical processing such as milling or etching). The open channel thus created is then closed by a separately produced sensor layer 7.

[0146] Figure 6e shows another embodiment of a microchannel 5 with a rectangular cross-section, wherein the microchannel 5 is first formed on the surface of the base material of the quantum magnetic field sensor 6.

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[0150] -18- was formed. The open channel created in this way is then closed by a separately manufactured cover 13. The sensor layer 7 is formed in the base material of the quantum magnetic field sensor 6 and forms the groove base of the microchannel 5.

[0151] Figure 6f shows an embodiment of a microchannel 5 with a semicircular cross-section, wherein a cover 13 with a semicircular inner shape covers the sensor layer 7 of the quantum magnetic field sensor 6.

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[0157] REFERENCE MARK LIST

[0158] 1 cell of a first cell type

[0159] 2 cells of a second cell type

[0160] 3 cells of a third cell type

[0161] 5 microchannel

[0162] 6 Quantum magnetic field sensor

[0163] 7 sensor layers with color centers

[0164] 8 flow regulators

[0165] 9 sorting device

[0166] 10 microfluidic switches

[0167] 11 Magnetic nanoparticles of the first kind

[0168] 12 Magnetic nanoparticles of the second kind

[0169] 13 lids

Claims

DELETE FAU Godfather Lawyers 251021 -WO -20- PATENT CLAIMS 1. Method for detecting cells of a first cell type (1) in a 5. Biopsy sample containing a plurality of cells of different cell types (2, 3), with the following procedural steps in the specified order: Performing tissue dissociation of the biopsy sample to isolate the cells contained in the biopsy sample; Supply of magnetic nanoparticles (11 ) to the isolated cells for selective labeling of the cells of the first cell type (1 ); Removal of the superfluous magnetic nanoparticles (11 ); Introducing the cells into a microchannel (5), wherein the microchannel (5) has a cross-section that allows the simultaneous passage of only one labeled cell (1); Performing a magnetic field measurement in the microchannel (5) using a quantum magnetic field sensor (6) based on the color center principle; and Sorting out the marked cells (1) after passing through the microchannel (5).

2. Method according to claim 1, wherein the quantum magnetic field sensor (6) has a sensor layer (7) in which color centers are formed and which forms at least a partial area of ​​an inner surface of the microchannel (5).

3. Method according to one of claims 1 or 2, wherein the magnetic nanoparticles (11) are designed as superparamagnetic iron oxide nanoparticles. DELETE FAU Godfather Lawyers 251021 -WO -21- 4. Method according to one of claims 1 to 3 further comprising the following method step: changing the cross-section of the microchannel (5) depending on the size of the cells.

5. Method according to any one of claims 1 to 4, wherein the biopsy sample is a fine needle biopsy sample.

6. Method according to any one of claims 1 to 5, wherein the magnetic field measurement is a continuous wave measurement.

7. Method according to any one of claims 1 to 5, wherein the magnetic field measurement is performed as a pulsed measurement.

8. Method according to any one of claims 1 to 7, wherein after the 5th process step “sorting out the marked cells” a further process step is carried out as a light microscopic examination of the sorted, marked cells (1 ).

9. A method according to any one of claims 1 to 8, wherein in the process step "addition of magnetic nanoparticles", first magnetic nanoparticles (11) are added for the selective labeling of cells of the first cell type (1) and second magnetic nanoparticles (12) are added for the selective labeling of cells of a second cell type (2), and wherein in the process step "sorting out the labeled cells", the cells (1, 2) labeled with the first magnetic nanoparticles (11) and those labeled with the second magnetic nanoparticles (12) are sorted out.

10. A method according to any one of claims 1 to 9, wherein the cells remaining after the process step "sorting out the labeled cells" are DELETE FAU Godfather Lawyers 251021 -WO -22- The cells are again subjected to the process steps "addition of magnetic nanoparticles" to "sorting out the labeled cells", whereby in the process step "addition of magnetic nanoparticles" further magnetic nanoparticles are then added for the selective labeling of cells of another cell type.

11. Device for performing a magnetic field measurement on cells (1) of a biopsy sample labeled with magnetic nanoparticles (11), comprising: a microchannel (5) wherein the microchannel (5) has a cross-section that allows the simultaneous passage of only one labeled cell (1); a quantum magnetic field sensor (6) based on the color center principle for performing a magnetic field measurement in the microchannel (5).

12. Device according to claim 11, wherein the The quantum magnetic field sensor (6) has a sensor layer (7) in which color centers are formed and which forms at least a part of an inner surface of the microchannel (5).

13. Device according to one of claims 11 or 12, wherein the microchannel (5) has a variable cross-section.

14. Device according to one of claims 11 to 13 with a flow regulator (8) at the inlet of the microchannel (5). DELETE FAU Godfather Lawyers 251021 -WO -23- 15. System for detecting cells of a first cell type (1) in a biopsy sample containing a plurality of cells of different cell types, comprising: a tissue dissociation device for performing a 5 Tissue dissociation of the biopsy sample for isolating the cells contained in the biopsy sample; a labeling device for supplying magnetic nanoparticles (11) to the isolating cells and for selectively labeling the cells of the first cell type (1); 0 a removal device for removing the excess magnetic nanoparticles (11); a device according to any one of claims 11 to 14; and a sorting device (9) for sorting out the labeled cells (1) after passing through the microchannel (5).

16. System according to claim 15, wherein the sorting device (9) comprises a microfluidic switch (10) arranged at the output of the microchannel (5).

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