Such methods are used, in particular, in chemical, biochemical, biological or biomedical laboratory environments to determine the concentration of chemical, biochemical, biological or biomedical analytes in sample solutions. A typical principle of such optical meas- urements is based on the measurement of fluorescent light. The analyte is marked by a fluorescence marker, which can be excited by the excitation light of a light source. The fluorescence marker receives the light and emits a sample light, typically at a different wavelength (fluorescence). The intensity of the detected sample light is used to derive the concentration of the analyte in the solution. Ideally, the concentration of the analyte is proportional to the detected intensity and can be determined using a calibration factor. Such a calibration factor can be predetermined, e.g. contained in a dataset in the memory of a laboratory apparatus, or can be determined by conventional calibration methods. Laboratory apparatus, which use the measurement of fluorescence light are, for example, fluorometer, which determine the intensity of fluorescence sample light, flu- orescence spectrometers, which determine the intensity of fluorescence sample light in dependence on the wavelength of the excitation light, or realtime PCR instruments, which monitor the progress of a polymerase chain reaction (PCR) by observing the fluorescent light from the PCR samples over the time.

Additional technical effort for performing the optical measurement is required in the case that it is desired to measure multiple analytes, which have substantially different concentrations. The electric photodetectors, which are typically used as detector devices in laboratory apparatus for performing optical measurements, are limited to specific ranges of detectable light intensities for a predetermined set of operational parameters of the laboratory apparatus and, in particular, the detector device. Intensities within said ranges can be detected without changing the set of operational parameters. For measuring intensities outside from said range, the set of operational parameters of the detector device has to be varied. Typically, electric gain is used to vary the sensitivity of a detector device. Alternatively, the intensity of the excitation light is varied to simulate a variation of the sensitivity parameter of the laboratory apparatus. Many commercially available detector devices are designed to automatically provide or to use a sensitivity parameter, which is suitable to measure the intensity of the incident sample light. The detector de- vice automatically determines the - more or less correct - intensity in regard to the sensitivity parameter, which was applied during the measurement. In an ideal case, apart from noise and signal saturation or overload, the intensity I is proportional to the value of the sensitivity parameter S, at least in a substantially linear area of the function l(S). I(S) depends, as a rule, also on the wavelength, at which the sample light is detected by the detector device.

Anyway, the detector devices can use internal corrections to apply the correct relations between intensity and sensitivity. The correction may by valid for the detector device alone, but may need further modification when the detector device is part of a laboratory apparatus. In a laboratory device, the combination of a detector device with the meas- urement setup of a laboratory devices, including the light sources, filters, means for guiding the optical path will lead to a situation, where the use of different sensitivity parameters leads to a reduced accuracy of the intensities evaluated by the detector device. In particular, the accuracy of a comparison of the intensities, which are detected at different sensitivity parameters, is reduced, because a precise calibration of the whole laboratory apparatus is missing.

One possibility of increasing the accuracy of a comparison of the intensities, which are measured for different analytes, is to use only a single value of the sensitivity parameter for the operation of the detector device. In this case, the range of intensities, which can be detected by the detector device, is limited by noise effects and signal saturation ef- fects such that a small range only is available for the measurement. Moreover, is a disadvantage that the optimal sensitivity parameter has to be found before, for example manually by the user.

It is the object of the present invention to provide a method for quantitative optical measurements, which provides an increased range for measuring light intensities, which can be compared with sufficient accuracy. It is a further object of the present invention to provide a laboratory apparatus, which utilizes the method according to the invention. The object is met by the method according to claim 1 and the laboratory apparatus according to claim 13. -Preferred embodiments of the- invention are subjects of the -sub claims.

The method according to the invention is a method for the quantitative optical measurement of a characteristic property of at least one analyte in at least one laboratory sample, in particular for the fluorescence measurement of at least one biochemical or biological sample, the method using a laboratory apparatus, which has at least one light source and at least one detector device, the apparatus utilizing at least sensitivity parameter S, which controls the capability of the laboratory apparatus to detect a signal by means of the at lest one detector device, the method using source light for causing the at least one sample to emit a sample light, and the at least one detector device for detecting sample light and the method utilizing the at least one sensitivity parameter S to detect the corresponding at least one intensity I of the sample light, preferably utilizing at least two different sensitivity parameters S and detecting the corresponding at least two intensities I of the sample light, the method comprising the steps:

- determining at least one reference point (S ref; I ref) ;

using at least one first sensitivity parameter S_m1 , which is not the same as S_ref, for measuring at least one first intensity I_m1 of sample light assigned to a first analyte;

- determining a quantity Q1 , which is a measure for the slope of a line, which is determined by utilizing the at least one reference point (S_ref; l_ref) and the at least one measurement point (S_m1 ; I_m1);

- using the quantity Q1 for calculating a first analyte value C_m1 , which is dependent on Q1 and which is characteristic for a property of the first analyte, in particular for a concentration of the first analyte in the at least one sample. In a preferred embodiment of the method, the quantity Q1 is the slope of a line, preferably a straight line, which is defined at least by the two points (S ref; I ref) and (S_m1 ; I_m1), Q1 being preferably calculated according to the formula

Q1 = (I_m1 - l_ref) / (S_m1 - S_ref), or a€<;ording-to an interpolation function or— line-or a regression function or- -line through— (S_ref; l_ref) and multiple different measurement points (S_m1 ; I_m1), and wherein, in particular, C_m1 is proportional to Q1 , preferably C_m1 = Q1. Here, the reference point, preferably, is a standardization point, which preferably is the same for at least one analyte (or at least two analytes), which, in particular, is measured utilizing at least two different sensitivity parameters S. Furthermore, the reference point, preferably, is the same for at least one predetermined characteristical wavelength of detection. Pref- erably, one standardization point is used for each characteristical wavelength of detection.

In another preferred embodiment of the method, the quantity Q1 is the slope of a straight interpolation line or regression line, which is adapted to (S_m1 ; I_m1) and multiple refer- ence points (S_n1 , I_n1), which are determined using the same first analyte.

Preferably, the relation l(S) is considered to by dependent also on the characteristical wavelength of the sample light, which is detected by the detector device. Typically, the at least one sample is illuminated with source light, which has at least one characteristical wavelength co_s, which is the wavelength, where the source light intensity l(co_s) has a maximum. The source light can be monochromatic, which means, it emits light substantially only at the characteristical wavelength. The detector device can be equipped with optical filters to detect only one or more predetermined spectral region(s) around a characteristical wavelength ω of detection, which preferably corresponds to the characteristi- cal emission wavelength, at which the spectrum of sample light has a maximum. This emission wavelength can be defined by the fluorescence of an analyte. Preferably, the method comprises the step to let the detector device detect the intensity of the sample light in dependence on at least one predetermined characteristical wavelength of detection, and wherein the at least one reference point(s) (S_ref; I ref) is respectively deter- mined in dependence on the characteristical wavelength of detection, thus assigning the at least one reference point(s) (S ref; I ref) to each characteristical wavelength of detection.

The method is applied, preferably, in chemical, biochemical, biological or biomedical la- boratories to determine a characteristic property of at least one analyte in at least one sample. Preferably, the characteristic property is the concentration of an analyte, preferably a -chemical, biochemical^ biological or -biomedical analyte, -in a sample, preferably a chemical, biochemical, biological or biomedical sample. Typically, a plurality of different samples are provided separately, each sample having at least one analyte or exactly one analyte to be optically measured, preferably by a fluorescence measurement. It is also possible and preferred, that one sample is measured, which has two or more analytes to be measured. A laboratory apparatus for performing the method of the invention can be configured to hold one sample, which, for example, can be exchanged by the user by another sample to be measured, or to hold multiple samples, for measuring multiple samples arranged in the laboratory apparatus, in parallel or sequentially.

Preferably, the method provides that a specific sensitivity parameter is chosen, prefera- bly automatically, preferably semi-automatically or preferably manually, for a specific concentration of the analyte in the sample, such that the sensitivity parameter allows to operate the detector device in a linear range, where the measured intensity I is substantially linear to the sensitivity parameter S. Typically, each sample with each analyte is measured with a specific sensitivity parameter.

The use of a reference point (S_ref; l_ref) for deriving C_m1 , in particular the concentrations of the analytes in the samples, increases the accuracy when comparing -or further calculating - the values of the concentrations. Instead of deriving the concentration, e.g. the concentration CON1 of a first analyte, just from one measurement, e.g. according to CON1 = b, b being a calibration function b( I_m1 ) or CON1 = b(l_m1 - I_m0), and I_m0 being a measurement with a blank sample without analyte, according to the invention the concentration is derived from the slope C_m1 of a straight line, which runs through the reference point and the measured point (S_m1 ; I_m1) or which is a regression line through multiple reference points and the measured point.

Preferably, the reference point (S ref; l_ref) is a standardization point (S fix; I fix) =

(S_ref; l_ref) and the step of determining a standardization point (S fix; l_fix) comprises the steps: • using the source light and at least one optical standard sample for letting the — at-least one-standard sample emit at least one standard sample light, and letting the detector device detect the at least one standard sample light for determining the first intensity M of a first standard sample light as a first function of the variable sensitivity parameter S, according to l_1 (S) = c1 ^* (S - S_1_0)

+ l_1_0, wherein c1 is a factor depending on the first optical standard and

S_1_0 and I 1 0 are the parameters for defining the straight line, which can be determined using the dataset (S; 1 1);

• utilizing at last one second function l_2(S);

· determining (S_fix; l_fix) to be an intersection point between the at least two functions l_1 (S) and l_2(S).

In this case, a single standardization point (S_fix; l_fix), which preferably is determined prior to the measurement of the samples, is used for the measurement of different ana- lytes, preferably in different samples, preferably for least at one predetermined character- istical wavelength of detection of the detector device, wherein preferably one individual standardization point (S_fix; l_fix) is assigned to one characteristical wavelength of detection. I_1 (S) is determined by variation of S and detecting the associated intensity 1 1 (S). In order to determine S_1_0 and l_1_0, the dataset (S; M) should contain at least two points (S; M). Preferably, the dataset (S; l_1) contains a plurality of points (S; M). The method of determining M (S) can comprise a conventional regression or interpolation method for fitting a function l_1 (S) to the measured pairs of values (S; l_1). The standardization point (S_fix; I fix) can be shared by all calculations C_m1 , C_m2, C_m3,

C_m4, ... C_mN of a measurement of a number of N (N>0) analytes. This further im- proves the overall accuracy of a comparison of different intensities, which are related to measurements at different sensitivity parameters S_m1 , S_m2, S_m3, S_m4, ... S_mM, wherein M can be a number, which preferably is smaller than or equal to N.

Preferably, an optical standard sample is an optical sample, which is predetermined and representative for a certain concentration of an analyte, in particular representative for a certain concentration of a fluorescence marker. Usually, optical standard samples are available as a set of samples, wherein an optically interacting (e.g. by fluorescence) substance is contained in each sample with a different predetermined concentration. However, standard samples can be prepared by the user, e.g. by dissolving a sample analyte in a solution at different predetermined concentrations. The parameter c1 can be determined using a conventional calibration method.

Preferably, the method according to the invention comprises the further steps: using at least one second sensitivity parameter S_m2 for measuring at least one second intensity I_m2 of sample light assigned to the first analyte, or preferably, to a second analyte;

determining a quantity Q2, which is a measure for the slope of a line, which is determined by utilizing the same or another standardization point (S fix; I fix) and the at least one measurement point (S_m2; I_m2);

using the quantity Q2 for calculating an analyte value C_m2, which is dependent on Q2 and which, preferably is the second analyte value, which is assigned to the second analyte, and which is characteristic for the property of the analyte, preferably the second analyte, in particular for a concentration of the analyte, preferably the second analyte, in the at least one sample, in particular according to the formula

Q2 = (I_m2 - I fix) / (S_m2 - S fix) or according to a regression line through (S_fix; l_fix) and multiple different measurement points (S_m2; I_m2). and wherein, in particular, C_m2 is proportional to Q2, in particular, C_m2 = Q2. This applies, in particular, if more than one analyte is measured, for example, if at least two analytes are measured. In this case, in particular, the method according to the invention preferably comprises the further step of comparing the first sample value C_m1 and the second sample value C_m2 for determining a comparison value. A comparison can be performed, for example, by calculating a difference, for example, for receiving a result which is proportional to (C_m1 - C_m2). Further, a comparison can be performed, for example, by calculating a division operation, for example, for receiving a result which is proportional to (C_m1 / C_m2). Other comparison operations are possible. In particular, C_m2 can be measured using also the first analyte. In this case, the measurement of the same first analyte at two different sensitivity parameters can be used to determine, for example, an average quantity <C_m>, which can be <C_m> = 0.5 ^* (C_m1 + C_m2). This can improve the accuracy of the result of determining the property of the first ana- lyte, e.g. the-concentration _:

In the method of determining a standardization point, the second function l_2(S) can be l_2(S) = constant, for example l_2(S) = 0. This can achieve sufficient accuracy for a comparison operation of different intensities, measured for different analytes at different sensititvity parameters.

Preferably, in the step of determining a standardization point, at least two optical stand- ard samples are used, for additionally determining the second intensity l_2 of at least also the second standard sample light as the second function l_2(S) of the variable sensitivity parameter S, according to l_2(S) = c2 * (S - S_2_0) + \_2_0, wherein c2 is a factor depending on the second optical standard, and which can be received by conventional calibration methods. S_2_0 and l_2_0 are the parameters for defining the straight line, which can be determined using the dataset (S; l_2). I_2(S) is calculated by variation of S and detecting the associated intensity l_2(S). The method of determining l_2(S) can comprise a conventional regression or interpolation method for fitting a function l_2(S) to the measured pairs of values (S; l_2). In many cases, the use of two straight lines 1_1 (S) and l_2(S) has proved to achieve sufficient accuracy for a comparison operation of dif- ferent intensities, measured for different analytes at different sensititvity parameters.

Preferably, the method of determining a standardization point comprises the steps:

• utilizing at least three optical standards to determine three intensities of at least three standard sample lights as the functions l_1 (S), l_2(S) and l_3(S);

• determining (S_fix; i fix) to be an estimated intersection point between the at least three functions l_1 (S), l_2(S) and l_3(S), using an mathematical estimation method . Here, additionally determining the third intensity l_3 of at least also the third standard light as the third function l_3(S) of the variable sensitivity parameter S, according to l_3(S) = c3 * (S - S_3_0) + l_3_0, wherein c3 is a factor depending on the third optical standard sample, and which can be received by conventional calibration methods. S_3_0 and l_3_0 are the parameters for defining the straight line, which can be determined us- ing the dataset (S; l_3). The method of determining l_3(S) can comprise a conventional regression or interpolation method for fitting a function l_3(S) to the measured pairs of values (S; l_3).

In case that three or more than three functions l_1 (S), l_2(S) and l_3(S) are used for determining (S_fix; l_fix), it is likely that the three or more function do not have a single intersection point. Probably, three intersection point of pairs of functions will exist. In this case, any mathematical method can be used, preferably, which defines the intersection point to be an estimated intersection point. The mathematical estimation method can be a geometrical method, or a method which averages the coordinates of the intersection points of pairs of functions, or any other regression method.

In most cases, the use of at least three straight lines l_1 (S), l_2(S) and l_3(S) has proved to efficiently achieve optimal accuracy at reasonable effort for a comparison operation of different intensities, measured for different analytes at different sensititvity pa- rameters.

Preferably, the mathematical estimation method provides the following steps:

• determining (S_fix; l_fix) to be a point within the area, which is enframed by the at least three functions l_1 (S), l_2(S) and l_3(S).

In most cases, this choice of (S_fix; l_fix) is most efficient and shows sufficient accuracy for a comparison operation of different intensities, measured for different analytes at different sensitivity parameters.

Preferably, at least three or exactly three optical standard samples are used to determine three straight line functions l_1 (S), l_2(S) and l_3(S), and wherein the mathematical estimation method provides the following steps: · determining the three intersection points IS1 , IS2, IS3 of the three pairs of straight lines (l_1 (S); l_2(S)), (l_2(S); l_3(S)), and (M (S); l_3(S));

• determining (S_fix; l_fix) to be the intersection point of the bisecting lines of the three angles of the triangle defined by the three intersection points IS1 , IS2, IS3. This choice of (S fix; I fix) is very efficient and shows improved accuracy for a comparison operation of different intensities, measured for different analytes at different sensitive ty parameters. Instead of using a common standardization point, which is used for all measurements C_m1 , C_m2, C_m3, C_m4, ... C_mN with different sensitivity parameters S_m1 , S_m2, S_m3, S_m4, ... S mM, preferably at least for one characteristical wavelength of detection, it is the approach in another preferred embodiment method according to the invention to multiple times use one analyte in a sample for more precisely determining each value C_m1 , C_m2, C_m3, C_m4, ... C mN. In this case, in particular, the method according to the invention comprises the modified steps of determining at least one reference point (S ref; \_ref)

• using at least one further sample light assigned to the first analyte for letting the detector device detect at least one reference light from said sample, by determining a dataset with pairs of the first intensity 1 1 of the first reference light in dependence from the variable sensitivity parameter S;

• utilizing at least one pair (S; l_1 ) of the dataset to determine the at least one reference point to be (S_ref; l_ref) = (S; l_1).

The dataset can comprise two ore more points (S_mi; l_mi). The value C_m1 can be derived from the slope of the straight line, which runs through two points (S_m1 ; I_m1) and (S_m2; I_m2) of measurements of the first analyte. This method has shown to have a higher accuracy than determining C_m1 = d * I_m1 , d being a calibration factor or function.

The value C_m1 can further be derived from the slope of the straight line, which runs through multiple points (S ni; I ni) of measurements of the first analyte: preferably, multiple reference points are determined using the detected intensities I_n1 of the sample light assigned to the first analyte at different sensitivity parameters S_n1 , wherein a conventional regression method is used to define a regression line through the measured point (S_m1 ; I_m1) and the multiple reference points (S_n1 ; I_n1 ), and wherein the first analyte value C_m1 is derived from the slope c_i of the regression line, wherein C_m1 is in particular proportional to c_i. Preferably, the first analyte value C_m1 is used to determine the concentration of the first -analyte in the-at least-one sample, according to CON1 = a(C_m1) or CON1- = a (GHTI4 C mO), wherein a is a predetermined function or a constant), which is in particular determinable using a conventional calibration method, and wherein C mO is the value of a blank sample, which does not contain the analyte. C_m0 can be determined by performing a measurement of the background radiation, e.g. a background fluorescence measurement, using the laboratory apparatus for performing the method, which contains the same sample holder, sample receptacle and preferably the solution, which solution is the same used for solving the analyte. The sample can be defined as the analyte with the solution, in which the analyte is solved. Preferably, CON1 is proportional to C_m1 or to (C_m1 - C_m0).

The sensitivity parameter can be understood to be any operational parameter of a laboratory apparatus, which has a detector device, which parameter controls the capability of the laboratory apparatus to detect a signal by means of a detector device. Usually, "sensitivity" of a sensor is defined as an output voltage change for a defined change in input. Preferably, the sensitivity parameter is varied by variation of at least one operational parameter of the detector device, e.g. an electronic gain factor of an electrical detector device, e.g. a photometer. It is also possible and preferred that the sensitivity parameter is varied by variation of the intensity of the source light, which is used to measure the at least one analyte in the at least one sample. Also in this case, the value of the detected signal is increased, which means that the effect of increasing the intensity of the light source is equivalent to increasing the sensitivity of a sensor. The laboratory apparatus according to the invention for quantitative optical measurements of a characteristic property of at least one analyte in at least one laboratory sample, in particular fluorescence measurements of at least one biochemical or biological sample, comprising at least one light source for illuminating the at least one sample with source light, at least one detector device for detecting sample light, and an electric con- trol device, wherein the electric control device is configured to at least automatically calculate in dependence on Q1 , using the method according to the invention or one of its preferred embodiments, the first analyte value C_m1 , which is characteristic for a property of the first analyte, in particular for a concentration of the first analyte in the at least one sample. Preferably, the method according to the invention is implemented by the apparatus or the control -device, preferably by utilizing a eemputer-program code, whiert preferably is stored on a memory device, e.g. optical data disc or a flash memory. Preferably, a laboratory apparatus according to the invention is configured to store said computer program code in a data memory of the laboratory apparatus. Preferably, said computer program code is executable and the laboratory apparatus according to the invention is configured to execute the computer program code for applying the method according to the invention. Preferably, the laboratory apparatus according to the invention is configured to be a fluo- rometer, or a fluorescence spectrometer, or a realtime PCR instrument. Other preferred method steps can be derived from the description of the functions of the laboratory apparatus according to the invention. The laboratory apparatus preferably comprises other components, e.g. a sample holder area for accommodating a sample holder, for holding at least one sample vessel, in particular for holding single vessels, e.g. sample tubes, capped or uncapped, or for holding multiple sample receptacles, e.g. microtiter plates or PCR-plates, user interfaces, e.g. a keyboard, display or a touchscreen, data exchange interfaces and connections. The la- boratory apparatus, in particular the sample holder, can comprise temperature devices, e.g. Peltier elements, for heating and/or cooling the sample holder, and/or thermosen- sors. The laboratory apparatus and the electric control device can comprise electric control loops for controlling the temperature of the sample holder or the samples, respectively. Preferably, the laboratory apparatus is configured to apply a temperature program to the sample holder, e.g. for repeatedly adjust the temperature of the sample holder through different temperature levels (thermal cycling). Preferably, the laboratory apparatus is configured to be a thermocycler for performing realtime PCR.

A preferred use of the method according to the invention or the laboratory apparatus ac- cording to the invention is to optically measure the fluorescence light of biochemical samples or biological samples, in particular PCR samples.

Moreover, further advantages, features and applications of the present invention can be derived from the following embodiments of the method and the laboratory apparatus ac- cording to the present invention with reference to the drawings. In the following, equal reference- signs substantially describe equal-devices.

Fig. 1 schematically shows an embodiment of a laboratory device according to the invention, which is configured to apply the method according to the invention.

Fig. 2 schematically shows a flow diagram of an embodiment of the method according to the invention.

Fig. 3a, 3b, 3c and 3d show, respectively, a diagram with the curves, which are used to determine a standardization point in step 11 of the method according to Fig. 2.

Fig. 4 shows a table, which can be used by embodiments of the method and the laboratory apparatus according to the invention, which table assigns a sensitivity parameter to the detected intensity of sample light, according to step 16 of the method in Fig. 2.

Fig. 5 schematically shows how the concentration of the analyte in the sample is derived from the measured point and the standardization point, according to step 17 of the method in Fig. 2.

Fig. 6 schematically shows a flow diagram of another embodiment of the method according to the invention.

Fig. 7 schematically shows how the concentration of the analyte in the sample is derived from the measured point and the standardization point, according to step 23 of the method in Fig. 6.

Fig. 1 a laboratory device 1 according to the invention, which is configured to apply the method 10, which is shown in Fig. 2. The laboratory apparatus is a fluorescence spectrometer 1 for performing fluorescence measurements of multiple biochemical or biological samples, e.g. PCR-samples. The laboratory apparatus has a light source 2, which is configured to emit light at one excitation wavelength of 470 nm, in the case of the embodiment, in particular to emit the light 3 to a first sample 5 within a sample holder 4, and to subsequently or simultaneously emit light 3' to each analyte in each sample of the sample holder. The embodiment of the laboratory apparatus works in the same way, if only one sample receptacle is provided, which contains two reagents. The laboratory -apparatus further comprises the-detector-device 7, which has at least one photodetector 8 for detecting the sample lights 6, 6', and for providing electrical signals, which quantify the intensity of a sample light 6, 6'. The detector device 7 is a commercial FluoSens- module, QIAGEN GmbH, Germany. The sensitivity parameter of the FluoSens-module can be changed by controlling the intensity of light sources (LED which emit the excitation light). The photodetector has two characteristical wavelengths of detection, which are 520 nm and 560 nm, in the embodiment.

Moreover, the detector device or the laboratory apparatus, respectively, have an electric control device 9. The electric control device 9, preferably, comprises a microprocessor, or at least a CPU and memory, in particular RAM and solid memory. The electric control device is configured, preferably, for applying the method according to the invention, in particular the method 10 or 20. This means that, for example, the electric control device comprises programmable circuits, which are programmed to perform the method according to the invention. Said programming can be a software programming, which uses a computer program code.

Fig. 2 shows a flow diagram of the method 10 for the quantitative fluorescence measurements of at least one biochemical or biological sample 5. The typical task is to determine the concentrations of multiple analytes in multiple separate samples or one single sample and to accurately compare the determined concentrations. The method 10 is suitable to achieve a sufficient accuracy of such a measurement with subsequent comparison of the detected concentrations of the analytes. The method 10 is run automatically by the laboratory apparatus 1 and provides said accurate measurements without any substantial additional user interaction, which is comfortable for the user and increases the efficiency of the work flow in a laboratory.

Initially, the method 10 provides an adjustment step 11 of the laboratory apparatus. Said adjustment provides the use of three optical standard samples, e.g. user prepared standard samples, for determining the reference point (S ref; I ref) in step 11 , which is a standardization point (S_fix; l_fix). The single standardization point (S_fix; l_fix) is used to accurately determine the measured quantities C_m1 ... C mN of multiple analytes, using multiple sensitivity parameters S_m1 ... S_mM. In case of the present embodiment, two standardization points (S fix; I fix)-1 and (S fix;

ΜΊχ)-2 are-determined (not shown)-, wherein (S^fix;- l_fix)-1-is- determined using-the characteristical wavelength of 520 nm of the detector device for detecting only light at 520 nm (i.e. within a rather narrow spectrum around 520 nm), and (S_fix; l_fix)-2 is de- termined using the characteristical wavelength of 560 nm. The following steps are described for the measurement at 520 nm and have to be repeated for measuring at 560 nm, in the embodiment.

In step 12 of the determination of the standardization point (S fix; I fix) , a first dataset of multiple points (S_1 ; l_1) is determined, at the characteristical wavelength of 520 nm. This means that for the first optical standard sample a predetermined light intensity is illuminating the samples, and the excited sample light is detected by the detector device, while using multiple (at least two) different sensitivity parameters S_1. For each value of S_1 , the intensity M of the sample light is detected, measured and stored. The resulting dataset is graphically shown in the diagram of Fig. 3a. A first straight line I 1 (S) is fitted through the points of said dataset. Generally, two points of the dataset are sufficient to draw a line. Providing multiple points, however, increases the accuracy of the estimation method. Said estimation method can be conventional, using a conventional curve fitting method to fit a straight line through (or between) the points of the dataset, e.g. a fitting method using least squares approximation. Preferably, at least (and/or a maximum of) 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 20, 25, 30, 35, 40, 50 or a different number of points are used for determining the dataset, which efficiently defines the straight line. Preferably, additionally algorithms are provided for sorting out outliers, which occur due to noise or signal saturation.

In step 13 of the determination of the standardization point (S_fix; I fix) , a second da- taset of multiple points (S_2; l_2) is determined. This means that for the second optical standard sample the predetermined light intensity is illuminating the sample, and the excited sample light is detected by the detector device (at the same characteristical wavelengths), while using multiple (at least two) different sensitivity parameters S_2. For each value of S_2, the intensity l_2 of the sample light is detected, measured and stored. The resulting dataset is graphically shown in the diagram of Fig. 3b, in addition to the dataset (S_1 ; l_1). In step 14 of the determination of the standardization point (S_fix; I fix) , a third dataset of multiple-points (S-3; -I^S) is -determined . -T is means that for the third optical- standard- sample the predetermined light intensity is illuminating the samples, and the excited sample light is detected by the detector device (at the same characteristical wave- lengths), while using multiple (at least two) different sensitivity parameters S 3. For each value of S 3, the intensity l_3 of the sample light is detected, measured and stored. The resulting dataset is graphically shown in the diagram of Fig. 3c, in addition to the da- tasets (S_1 ; M) and (S_2; l_2).

Respectively, a straight line l_1 (S) and l_2(DS) is fitted to the dataset (S_1 ; l_1) and (S_2; l_2).

In step 15 of the determination of the standardization point (S_fix; I fix) , the three intersection points IS1 , IS2, IS3 of the three pairs of straight lines (l_1 (S); l_2(S)), (l_2(S); l_3(S)), and (l_1 (S); l_3(S)) are determined. The standardization point is determined to be the intersection point of the bisecting lines of the three angles of the triangle defined by the three intersection points IS1 , IS2, IS3. The size of the area of the triangle can be used to evaluate the quality factor of the method of determining the standardization point. The intersection point is determined numerically, e.g. by the electric control device of a laboratory apparatus. The "geometrical" method of determining the standardization point (S fix; I fix) to be the intersection point of the bisecting lines of the three angles of the triangle defined by the three intersection points IS1 , IS2, IS3 is shown in Fig. 3d.

In step 16 of the method 10, the sensitivity S_m1 is automatically determined, which is suitable to measure the intensity of the sample light, which is assigned to the first analyte to be measured. The measurement starts with a default value of S_m1 and determines I_m1. A table (e.g. the table in Fig. 4) can be used to check if the detected value of I_m1 is comprised by the ranges of intensities (Fig. 4,"lower limit"; "upper limit") which are comprised in the table, and to determine the associated value of S_m1 (Fig. 4, "resulting sensitivity") in the table, which is assigned to the range of intensities comprising the measured I_m1. If the measured value of I_m1 is not. comprised in the ranges of intensities according to the table, the step 16 automatically is repeated with another value of S_m1 , until a valid value S_m1 is found. The intensity I_m1 , which is associated to the valid value of S ml is stored. In step 17 of the method 10, the standardization point (S_fix; I fix) is utilized for calculat- ing-a first analyte-value ^{^} C^ml , which is^haracteristie-for a property of the fiFst analytej 4F,~ particular for a concentration of the first analyte in the at least one sample, according to the formula C_m1 = (I_m1 - I ref) / (S_m1 - S_ref). C_m1 is the slope of the straight line, which runs through the two points (S fix; I fix) and (S_m1 ; I_m1). This is graphically shown in Fig. 5.

In step 18 of the method 10, the value of the concentration CON1 of the analyte in the sample is calculated from C_m1 , preferably according to CON1 = a(C_m1 - C mO), wherein a is a factor or a function, which is predetermined by a conventional calibration method and C_m0 is the result of measurement of the background fluorescence of the optical system, which comprises at least the parts of the optical path in the laboratory apparatus, the sample holder, sample receptacle, and the solution without the analytes. A conventional calibration method can comprise the steps of providing a set of samples, which each contain a sample analyte in a predetermined concentration, and by recording the intensities, which correspond to the sample, in particular using a standard source light, which can have standard intensity, standard frequency, preferably using standard illumination and/or detection times and/or a standard output energy of the light source.

The method steps 16 to 18 can be repeated for multiple samples and multiple analytes, using the same or another standardization point (S fix; i_f ix) , for one of the characteristi- cal wavelengths of detection or for multiple wavelength, to receive accurate results of the measurements, which can be accurately compared by calculation steps, in particular.

In Fig. 6, the alternative embodiment of the method according to the invention is shown, namely the method 20. Instead of calculating the reference point (S_ref; I ref) to be a common standardization point, the reference point is calculated in step 21 using the sample with the first analyte, instead of utilizing an optical standard sample. The method is preferably run at only one wavelength of detection, but can also be run once for each characteristical wavelength of detection. Method 20 comprises the step 22 of using at Jeast one further sample light assigned to the first analyte for letting the detector device detect at least one reference light from the sample, by determining a dataset with the points (S_m1 ; I_m1) and (S_m2; I_m2) with pairs of the measured intensity I_m1 (I_m2)in dependence from the variable sensitivity parameter S_m1 (S_m2). Herein, the values S_m1 , S_m2 are determined such that "valid" values of I_m1 , I_m2 are detected, e.g. by using a table as shown in Fig. 4; step 22 utilizes (S_m2; I_m2) to determine the at least one reference point to be (S ref; l_ref) = (S_m2; I_m2). The two points are used, in step 23, ^o-deri & G^rnt as the -slope -of -the straight line, which runs-through-the -two points (S_m1 ; I_m1) and (S_m2; I_m2). This is graphically shown in Fig. 7. In step 24, the value of the concentration CON1 of the analyte in the sample is calculated from C_m1 , preferably according to CON1 = a(C_m1 - C_m0), wherein a is a factor or function, which is predetermined by a conventional calibration method and C mO the result of a measurement of the background fluorescence. In case that a more extended measurement time is acceptable, multiple reference points S_n1 ; I_n1) can be used to even more precisely define the straight line and to derive the slope using conventional regres- sion methods, which leads to a more precise measurement of CON1. The same method 20 can be applied to each analyte.