METHOD FOR CONTROLLING A THERMOCYCLER AND THERMOCYCLER

Description

[0001] The invention relates to a method for determining a temperature change rate for controlling the temperature control device of a thermal cycler. The invention also relates to a thermal cycler whose control system is configured to carry out such a method.

[0002] A thermal cycler is a laboratory device capable of sequentially adjusting the temperature of at least one laboratory sample to a predetermined temperature and maintaining it at that temperature for a specified period of time. This temperature control process is cyclical. This means that a predetermined temperature cycle, i.e., a sequence of at least two temperature levels, typically three, is repeatedly performed. This process is typically used to perform a polymerase chain reaction (PCR).

[0003] When performing DNA amplification using PCR, the user relies on the sample being tempered precisely and reproducibly over the course of a tempering schedule specified by the user in order to achieve optimal results.

[0004] The temperature cycle of a thermal cycler is specified by the user by setting a temperature plan, usually directly on the device's control panel. An example of such a cycle with setting options in Fig. 2a with reference to a screen display of an exemplary thermal cycler according to the invention. The user sets the level of each temperature step of a cycle and its holding time, as well as the total number of cycles to be completed. The thermal cycler control system ensures that the temperature control plan is implemented as precisely as possible according to these specifications. Usually not precisely determinable for the user, the device control system uses specific control parameters that are adapted to the hardware components used in the thermal cycler. In particular, Peltier elements, which are usually used in the thermal cycler temperature control devices as temperature control elements for controlling the sample block (thermoblock), have different performance values. The result of the temperature control is also influenced by all

[0005] Hardware and material parameters of the thermocycler are determined, which have an influence on the heat transfer, i.e. the heat supply and heat dissipation, from the liquid laboratory samples to the Peltier element.

[0006] Key performance parameters of a thermal cycler are the maximum heating rate and the maximum cooling rate at which a thermal block of the thermal cycler can be tempered. An example of such a performance value is shown in Fig. 2b and Fig. 2c specified. Another performance characteristic is the transient response when setting the temperature levels of the thermoblock using temperature control. These performance values ​​are typical for a thermocycler or a class of thermocyclers. The transient response is generally optimized to reach a temperature level quickly. The corresponding design of the temperature control is device-specific and unknown to the user. Values ​​for the maximum heating rate and the maximum cooling rate are usually specified by the manufacturers in the device specifications. The actual temperature curve at the thermoblock of the thermocycler or the temperature curve in the liquid samples, which ultimately determines the result of the temperature-dependent reaction sequences (e.g. PCR), depends not only on the user-defined temperature control plan but also on the aforementioned hardware-specific performance values.

[0007] Document US 2002072112 A1 discloses computer-controlled instruments for performing a polymerase chain reaction (PCR) using thermocycling. A sequence task is used to execute the contents of a user-defined program. It sequentially cycles through each setpoint in a cycle consisting of a ramp and soak segment and sends setpoint temperature messages to a PID controller (PID task), which in turn controls the temperature of the sample block.

[0008] The investigations on commercially available thermal cyclers underlying the invention have shown that the maximum temperature change rates or ramp rates are usually not reached or only briefly reached. Figur 3c As shown, knowledge of the maximum rate of temperature change, which is represented in the figure as the steepest straight line 2, does not allow direct conclusions to be drawn about the time interval of the temperature change, here referred to as Δt s_cool. In other words, the maximum differential 2 of the temperature curve in the range of the time interval is not equal to the difference quotient 1 in the time interval. The difference quotient in the time interval is also referred to below as the effective rate of temperature change or effective heating rate / effective cooling rate. The difference quotient, starting from a first time t1 (end of the time of the first temperature stage) and a second time t2 (beginning of the time of the second temperature stage) and a first temperature T1(t1) and a second temperature T2(t2), is defined as (T2(t2) - T1(t1)) / (t2 - t1).The investigations revealed that the isolated consideration of ramp rates specified by manufacturers is often only partially meaningful and can even lead to incorrect conclusions regarding the mathematical estimation of the actual PCR runtime for a particular thermal cycler. In some cases, the runtimes of certain thermal cyclers were significantly longer than expected based on the ramp rates specified in the manufacturers' technical specifications. Minor deviations in the time to completion of a PCR can generally occur due to environmental factors (e.g., room temperature, device placement, etc.). However, several repeat runs confirmed that such differences are only within a few seconds.It was concluded that the following aspects contribute significantly to the observed discrepancies: For different thermal cyclers, the maximum ramp rates specified in the technical manuals are achieved for different periods of time during the ramp process from one temperature to the next—but for certain thermal cyclers, they may only be achieved for a short time during each ramp phase. Temperature control modes or the reaction volume setting can also significantly influence the ramp behavior. This may even require re-optimization of a reaction after transferring a PCR system from one thermal cycler to another.

[0009] When performing a PCR, users usually ignore the hardware-specific parameters. Instead, they optimize temperatures (e.g., using a gradient) to achieve optimal yield. When migrating a PCR to a different class of thermal cycler, this yield usually decreases. This is not only due to the temperature difference compared to the original device, but also, in particular, to deviations in dynamic behavior, i.e., on the ramp and during settling. Many users avoid switching to a different device to repeat experiments, as they do not produce the same results despite the same programming and application of the same temperature plan. When replacing the device, e.g.,Due to a defect or the replacement of existing equipment – ​​especially when changing the equipment type – a complex requalification is required to reproduce previous PCR results with the modified hardware. Since the operating programs of the thermal cyclers offer only limited options for influencing the control program and the control of the hardware components, requalification may also require a complex adjustment of the control program.

[0010] The object underlying the invention relates to reproducing a temperature profile in a thermal cycler when a user-defined temperature control plan is reapplied, in particular in a situation in which the hardware-specific performance values ​​have changed, as is the case, for example, when changing the thermal cycler.

[0011] In a preferred embodiment of the invention, the object is to facilitate the user's transition from one thermal cycler to another with different performance values. To this end, the thermal cycler is designed in particular according to the definitions of the present invention.

[0012] The invention solves this problem by the method according to claim 1 and the thermal cycler according to claim 9 as well as by the program code according to claim 13.

[0013] Preferred embodiments of the invention are the subject matter of the subclaims and can also be taken from the description and the figures of the invention.

[0014] The invention is based, among other things, on the observation that the total runtime of the temperature control plan executed on the respective thermal cycler is easily determined by the user and is usually also recorded. The user pays attention to this runtime, as it is the key parameter when developing the temperature control plan for determining the total reaction time or for optimizing throughput. According to the invention, the user specifies a previously known temperature control plan and a previously known runtime for this temperature control plan.

[0015] The application of the invention increases the probability of successfully reproducing a temperature profile. In particular, the dynamic behavior, including in particular the ramp rates and transient response, of an original thermal cycler is replicated or simulated without requiring the user to perform significant experiments and enter numerous parameters.

[0016] The runtime of a temperature control plan executed on a thermal cycler can be used to determine the temperature change rates, in particular the heating and cooling rates, used by the thermal cycler's temperature control device to reach the respective temperature level. The runtime then includes at least one holding time for at least one temperature level of the temperature control plan, as well as at least one time interval during which the adjustment of this at least one temperature level is carried out depending on at least one temperature change rate.For example, if one assumes that a thermal cycler always uses the same cooling rate for cooling between the corresponding temperature levels and always uses the same heating rate for heating between the corresponding temperature levels, then the running time T of the temperature control plan consisting of repetitions of the same temperature cycle results from the time intervals Δt S_heat of heating and the holding times T S_heat at this higher temperature level as well as from the time intervals Δt S_cool of cooling and the holding times T S_cool at this lower temperature level: . T = ∑ S heat = 1 n T S heat + Δ t S heat + ∑ S cool = 1 m T S cool + Δ t S cool T = ∑ s heat = 1 n T s heat + Δ s heat r H + ∑ s cool = 1 m T s cool + Δ s cool r C

[0017] Here, r H is the temperature change rate (heating rate) to heat the temperature block by a temperature difference Δ S_heat, and r C is the temperature change rate (cooling rate) to cool the temperature block by a temperature difference Δ S_cool.

[0018] In the context of this application, unless otherwise stated, the terms heating rate, cooling rate, and temperature change rate refer to the effective heating rate, the effective cooling rate, and the effective temperature change rate, respectively, and not to extreme values ​​that occur briefly during temperature changes. For most commercially available thermal cyclers, the effective heating rate and the effective cooling rate are in a roughly constant relationship. This can be easily determined for known thermal cyclers and stored as a table. This table can be stored in a data storage device in the thermal cycler or method according to the invention. The heating rate is usually higher than the cooling rate. This relationship can be sufficiently accurately approximated for thermal cyclers in many cases using the specification r H = 2*r C . The effective cooling rate or effective heating rate can then be calculated from the running time as follows: T = ∑ s hea = 1 n T s heat + Δ s heat 2 r C + ∑ s cool = 1 m T s cool + Δ s cool r C T = ∑ s heat = 1 n T s heat + Δ s heat 2 1 r C + ∑ s cool = 1 m T s cool + Δ s cool 1 r C T = ∑ s heat = 1 n T s heat + 1 r C ∑ s heat = 1 n Δ s heat 2 + ∑ s cool = 1 m T s cool + 1 r C ∑ s cool = 1 m Δ s cool T = ∑ s heat = 1 n T s heat + ∑ s cool = 1 m T s cool + 1 r C ∑ s hea = 1 n Δ s hea 2 + ∑ s cool = 1 m Δ s cool T − ∑ s heat = 1 n T s heat − ∑ s cool = 1 m T s cool = 1 r C ∑ s heat = 1 n Δ s heat 2 + ∑ s cool = 1 m Δ s cool <menclose notation="box"> r c = ∑ s heat = 1 n Δ s heat 2 + ∑ s cool = 1 m Δ s cool T − ∑ s heat = 1 n T s heat − ∑ s cool = 1 m T s cool , r H = 2 r C < / menclose> , unter der Bedingung , dass T > ∑ s heat = 1 n T s heat − ∑ s cool = 1 m T s cool

[0019] This calculation or a calculation with a comparable result, in particular the calculation of the framed cooling rate and the heating rate, is preferably carried out by an evaluation program that can be executed on a computer or on the data processing device of a thermal cycler according to the invention. The temperature change rates are determined, in particular, assuming a medium transient response (standard). This is exemplified in Fig. 2d shown. This affects the input variables Δ S_heat , Δ S_cool , T S_heat and T S_cool (for all s). It can be approximately assumed that the time intervals for heating and cooling each contain a constant time span (settling time period), which takes into account the settling of the control loop to the respective level of the temperature step. The settling time period is determined by the time period between the presence of the constant temperature change rate and the presence of a temperature step to be set. Alternatively, the settling time period can also be determined for certain commercial thermal cyclers and stored in a table. In addition, a table can contain a preselection from a limited selection of typical control modes. The table can be stored in a data storage device in the thermal cycler or method according to the invention.Alternatively or additionally, the thermal cycler or method according to the invention can be configured to allow the user to change the constant value of the settling time period as a variable—particularly by inputting data via a user interface device of the thermal cycler and thus entering data. The user can thus easily correct the result by changing the settling behavior if the yield is incorrect during the process.

[0020] Temperature control of a thermal cycler's temperature control block, optimized for controlled settling, is generally known. Settling to the desired setpoint temperature for a temperature stage of the temperature cycle, in the case of heating, involves an overshoot of the temperature applied in the temperature control block to a maximum temperature value that lies above the setpoint, followed by an undershoot to a temperature value below the setpoint, before then changing back to a lower temperature value above the setpoint, and so on, until the setpoint is reached. The settling can then be characterized by the temperature difference between the maximum temperature of the overshoot (in cooling: the minimum temperature of the undershoot) and the duration of the overshoot until the setpoint is reached. With a rapid settling, this temperature difference and the duration are both small.In particular, the user can be given a preselection of a limited number of transient modes to choose from. Each such transient mode can be characterized by specific values ​​for the specified temperature difference and duration, respectively for heating and cooling, i.e. in particular by two pairs of values: Mode _X : (Temperature difference _X , Duration_ X ) Heating , (Temperature difference _X , Duration _X ) Cooling . Such modes can be offered to the user via a list selection on the display, for example under the labels "Fast", "Intermediate", "Standard", "Safe", as is the case in the exemplary embodiment of the . Figur 2d is provided. Details on the implementation of such temperature controls using "overshoot" are known, for example, from EP 0 488 769 A2, described therein as a "controlled overshoot algorithm." Temperature control can also be achieved by additionally observing the effect of the temperature control on the temperature of the liquid sample contained in the vessel inserted into the thermoblock of the thermocycler, as known, for example, from EP 1 452 608 B1.

[0021] The method according to the invention and / or the thermal cycler according to the invention can be set up in such a way that the user - in particular via a user interface device of the thermal cycler - can select a specific commercial thermal cycler TC X, e.g. via a list selection that can be shown and operated on a display or touchscreen of the user interface device. The thermal cycler or the method then has the additional information that the running time T specified by the user relates to the implementation of the temperature control plan on the thermal cycler of the type TC X. Since the method according to the invention and / or the thermal cycler according to the invention can access the tables in which the ratio r H / r C or the settling time is stored as a function of TC X, the calculation of the temperature change rates can take place automatically after the user has selected TC X.According to the invention, the method is used to determine at least one temperature change rate for controlling the temperature control device of a thermal cycler, wherein the controller temperatures a sample-receiving thermoblock of the thermal cycler for carrying out polymerase chain reactions in these samples according to a temperature control plan, during which temperature levels are changed over by changing the temperature at a temperature change rate, comprising the steps: . Providing temperature control plan data that determine the holding time and the temperature of at least one temperature level of the temperature control plan; providing runtime data that determine the runtime required to execute the temperature control plan on a thermal cycler; determining this at least one temperature change rate by an evaluation program using the temperature control plan data and the runtime data; providing the at least one previously determined temperature change rate for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate.

[0022] Preferably, the temperature control plan data determine at least a first hold time and a first temperature of a first temperature stage and at least a second hold time and a second temperature of a second temperature stage of the temperature control plan. Typically, a cycle of the temperature control plan also has three temperature stages if it is a PCR, so that the temperature control plan data also determine a third hold time and a third temperature of the third temperature stage. The first temperature is assumed to be higher than the second temperature, wherein, according to the temperature control plan, switching takes place between temperature stages by cooling starting from the first temperature at a first temperature change rate (cooling rate) and heating starting from the second temperature at a second temperature change rate (heating rate).

[0023] The evaluation program preferably uses the temperature control plan data and the runtime data to determine at least one first temperature change rate, which is used as the cooling rate for setting the second temperature level, and at least one second temperature change rate, which is used as the heating rate for setting the first temperature level. This at least one cooling rate and at least one heating rate are preferably provided for controlling the temperature control device of the thermal cycler.

[0024] During a cycle of the temperature control plan, there is in particular at least one time interval during which at least one constant temperature change rate is applied and which may also contain a settling time period which may be determined by the time period between the presence of the constant temperature change rate and the presence of a temperature level to be set, wherein during this time period a settling process is carried out by the control of the temperature control device of the thermal cycler, which is part of the temperature control of a thermal cycler, wherein the method comprises the step: Providing settling data containing information about at least one settling time period, wherein the transient data are also used in particular to determine said at least one temperature change rate by the evaluation program.

[0025] Preferably, the method includes the step: Providing these transient data by input from a user at a user interface device of a data processing device by means of which the evaluation program is executed.

[0026] The runtime may also include a latency interval during which, at the beginning of a temperature control plan, a heatable lid covering the temperature control block of the thermal cycler containing the samples during the polymerase chain reaction is initially set to a target temperature, whereby the runtime data also includes information about this latency interval.

[0027] The method according to the invention preferably serves to control the temperature control device of a thermal cycler, wherein the method for controlling the temperature control device preferably includes a method for determining at least one temperature change rate from runtime data and temperature plan data. The thermal cycler comprises the temperature control device for controlling the temperature of a thermal block containing the samples for carrying out polymerase chain reactions in these samples according to the temperature plan specified in the method according to the invention, and comprises an electronic control device configured to control the temperature control device using control parameters. The method for controlling the temperature control device of a thermal cycler comprises the steps of the method for determining at least one temperature change rate from runtime data and temperature plan data and the following steps: Using the at least one previously determined temperature change rate to determine control parameters that include this at least one temperature change rate and that determine a temperature control plan corresponding to the temperature control plan; controlling the temperature control device by means of the control parameters and the electronic control device in order to execute this temperature control plan using this at least one temperature change rate.

[0028] The method for controlling the temperature control device is, in particular, a method for controlling a first thermal cycler by simulating the temperature control behavior of a second thermal cycler, wherein the method for controlling the temperature control device includes the method for determining at least one temperature change rate from temperature control plan data and runtime data that characterize the temperature control behavior of the second thermal cycler. In particular, the first thermal cycler can be operated with a first maximum temperature change rate, which is a cooling rate or a heating rate, and the second thermal cycler can be operated with a second maximum temperature change rate, which is a cooling rate or a heating rate, wherein the first maximum temperature change rate is greater than or equal to the second maximum temperature change rate.In short, the first thermal cycler preferentially heats faster than the second thermal cycler. An example of such a thermal cycler, whose maximum effective heating and cooling rates are greater than most of other commercially available thermal cyclers, is the Mastercycler®< X50 from Eppendorf AG, Hamburg, Germany. The Mastercycler®< X50 heats at a maximum of 10 °C / s and cools at a maximum of 5 °C / s.

[0029] The first thermocycler comprises the temperature control device for controlling the temperature of a thermoblock containing the samples for conducting polymerase chain reactions in these samples according to the temperature control plan defined in the method for determining at least one temperature change rate, and an electronic control device configured to control the temperature control device. The method for controlling the temperature control device comprises, in particular, the steps of the method for determining at least one temperature change rate from temperature control plan data and runtime data and the following steps: Using the at least one previously determined temperature change rate to determine control parameters which include this at least one temperature change rate and which determine a temperature control plan corresponding to the temperature control plan; controlling the temperature control device by means of the control parameters and the electronic control device in order to implement this temperature control plan under

[0030] Using this at least one temperature change rate.

[0031] In particular, the at least one temperature change rate is smaller than the first maximum temperature change rate.

[0032] The invention relates to a thermocycler, in particular for carrying out polymerase chain reactions in laboratory samples, comprising: a temperature control device for controlling the temperature of a thermoblock containing the samples according to a temperature control plan, during which temperature levels are changed by changing the temperature on the thermoblock at a temperature change rate;an electronic control device which has a data processing device and which is configured to control the temperature control device in order to carry out the following steps: using temperature control plan data which determine the temperature control plan and runtime data which determine the runtime of the temperature control plan, and using the at least one temperature change rate previously determined according to the method for determining at least one temperature change rate from temperature control plan data and runtime data to determine control parameters which include this at least one temperature change rate and which determine a temperature control plan corresponding to the temperature control plan;Controlling the temperature control device by means of the control parameters and the electronic control device to execute this temperature control plan using this at least one temperature change rate;

[0033] The electronic control device of the thermal cycler is configured to carry out the method for determining at least one temperature change rate from temperature control plan data and runtime data, wherein the electronic control device is configured to execute an evaluation program by means of the data processing device of the electronic control device and is configured to carry out the following steps: Acquiring temperature plan data that determine the holding time and the temperature of at least one temperature level of the temperature plan; Acquiring runtime data that determine the runtime required to execute the temperature plan on a thermal cycler; determining this at least one temperature change rate by means of the evaluation program using the temperature plan data and the runtime data; using the at least one temperature change rate determined by the evaluation program for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate.

[0034] The data processing device of the electronic control device preferably has an interface device via which a data connection can be established with an external data processing device, wherein the method for determining at least one temperature change rate from temperature control plan data and runtime data has been carried out on this external data processing device in order to provide the at least one temperature change rate, wherein the data processing device of the electronic control device is configured to receive this at least one temperature change rate, also the temperature control plan data and / or the runtime data, via the data connection.

[0035] Preferably, the thermal cycler comprises a user interface device, wherein the electronic control device is configured to record the temperature control plan data entered by a user via the user interface device and to record the runtime data entered by a user via the user interface device.

[0036] The invention also relates to a program code which carries out the following steps when executed by means of a data processing device of an electronic control device of a thermal cycler according to claim 9: Acquiring temperature plan data, which is particularly entered by a user via a user interface device connected to the data processing device, which is part of the thermal cycler, and which determines the holding time and the temperature of at least one temperature level of the temperature plan; Acquiring runtime data, which is particularly entered by a user via a user interface device connected to the data processing device, which is part of the thermal cycler, and which determines the runtime used to execute the temperature plan on a thermal cycler; Determining this at least one temperature change rate by means of the evaluation program using the temperature plan data and the runtime data, which is executed by the data processing device;Providing the at least one temperature change rate determined by the evaluation program for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate; using the at least one previously determined temperature change rate to determine control parameters that include this at least one temperature change rate and that determine a temperature control plan; controlling the temperature control device by means of the control parameters and the electronic control device in order to execute this temperature control plan using this at least one temperature change rate.

[0037] The invention also relates to the use of the method for determining at least one temperature change rate from temperature control plan data and runtime data for controlling the temperature control device of a first thermal cycler by simulating the temperature control behavior of a second thermal cycler. This simulation assists the user in migrating from the older, less powerful second thermal cycler to the more powerful first thermal cycler.

[0038] A thermal cycler is a device capable of sequentially adjusting the temperature of at least one sample to a predetermined temperature and maintaining it at this temperature level for a specified holding time. This temperature control process is cyclical. This means that a predetermined temperature cycle, i.e., a sequence of at least two temperature levels, is repeatedly performed. This method is particularly useful for conducting a polymerase chain reaction (PCR).

[0039] A thermal cycler, in particular the treatment device of the thermal cycler, preferably comprises a thermal block. A thermal block is a sample holder made of a thermally conductive material, usually a metal-containing material or a metal, in particular aluminum or silver. The sample holder has a contact side, which is contacted by at least one heating / cooling device of the thermal cycler, in particular at least one Peltier element, preferably several, in particular six, Peltier elements.

[0040] The thermal cycler, in particular the treatment device of the thermal cycler, has a control device with at least one control circuit, to which at least one heating / cooling device is assigned as an actuator and at least one temperature measuring device as a measuring element. The control device regulates the temperature of a temperature stage. A heat sink of the thermal cycler serves to cool sections of the thermal cycler, in particular the Peltier elements.

[0041] The thermal cycler, in particular the treatment device of the thermal cycler, can have additional heating and / or cooling elements. Preferably, the thermal cycler, in particular the treatment device of the thermal cycler, has a timer device with which temporal parameters for setting the temperature cycle can be controlled. In a thermal cycler, the device-controlled treatment of the at least one laboratory sample corresponds to a temperature cycle treatment to which the at least one sample is subjected.Possible parameters, in particular program parameters, in particular user parameters, which are used to influence a temperature cycle treatment in a temperature control plan, define in particular the temperature of a temperature level, the holding time of a temperature level, the control of further heating and / or cooling elements, and / or the number of temperature levels or cycles, and / or at least one sequence parameter which influences or defines the sequence, in particular the sequence, of a temperature control program consisting of several steps.

[0042] The thermal cycler has an electronic control device. Within the scope of the present invention, a control device has or is a data processing device, in particular a central processing unit (CPU) for processing data and / or a microprocessor. The control device or a computing unit of the control device of a thermal cycler is preferably also configured for program-based control of the temperature control of the thermal block.

[0043] The data processing device preferably has a computing unit, in particular a CPU, and preferably at least one data storage device, in particular for volatile and / or permanent storage of data. The data processing device is preferably designed to establish a data connection to an external computer or laboratory device, in particular a thermal cycler, via an interface device.

[0044] The use of a thermocycler for the cyclical temperature control of laboratory samples, in particular for performing PCR in these laboratory samples, is a device-controlled treatment, i.e., in particular, an at least partially automated treatment. With a partially automated treatment, it is particularly possible for the treatment to be carried out in such a way that, after starting the treatment and before ending the treatment, at least one user input is made, with which the user can influence the ongoing treatment, in particular, for example, by answering an automatic query issued via a user interface device of the thermocycler, in particular by confirming or denying an input or making other inputs.In semi-automated treatment, it is particularly possible for the treatment to comprise multiple treatment steps, which are carried out automatically one after the other, and at least one treatment step that requires user input, particularly via a user interface device. Examples of such user inputs on a thermal cycler include the input of temperature control plan data, the input of the runtime, and / or optionally the input or selection of a thermal cycler TC x assigned to the entered runtime.

[0045] The device-controlled treatment is preferably a program-controlled treatment, i.e., a treatment controlled by a program. A program-controlled treatment of a sample is understood to mean that the treatment process essentially occurs by executing a plurality or multiplicity of program steps. Preferably, the program-controlled treatment is carried out using at least one program parameter, in particular at least one program parameter selected by the user. A parameter selected by a user is also referred to as a user parameter. Typical user parameters in a thermal cycler determine the temperature control plan, in particular the level and holding time of the temperature steps of a temperature control cycle, the total number of cycles, and, within the scope of the present invention, also the runtime of the temperature control plan, which is familiar to the user from previous thermal cycler applications of the same temperature control plan.The program-controlled treatment is preferably carried out using a digital data processing device, which can in particular be a component of the control device of the laboratory device. The data processing device can have at least one processor, i.e., a CPU, and / or at least one microprocessor. The program-controlled treatment is preferably controlled and / or carried out according to the specifications of a program, in particular a control program. In particular, with program-controlled treatment, essentially no user action is required, at least after the user-required program parameters have been recorded.

[0046] A program parameter is a variable that can be set in a predetermined manner within a program or subprogram, valid for at least one execution (call) of the program or subprogram. The program parameter is set, e.g., by the user, and controls the program or subprogram and causes data output depending on this program parameter. In particular, the program parameter and / or the data output by the program influence and / or control the control of the device, in particular the control of the treatment by means of the at least one treatment device.

[0047] A program parameter can be a program parameter required by the user. A program parameter required by the user is characterized by the fact that it is necessary for carrying out a treatment. Other program parameters that are not required by the user can be derived from the program parameters required by the user or made available in other ways, in particular can be optionally set by the user. The setting of a program parameter by a user is carried out in particular by displaying a selection of possible predefined values ​​from a list of predefined values ​​stored in the laboratory device, with the user selecting the desired parameter from this list and thus setting it. This applies, for example, to the selection of a TCx thermocycler, which the user assigns to a known runtime of a temperature control plan.It is also possible for this program parameter to be set by the user entering the value, for example, by entering a number corresponding to the desired value using a numeric keypad, or by the user continuously or incrementally increasing or decreasing a value until it corresponds to the desired value, thus setting the value. Other forms of input, such as voice control and / or gesture control, are conceivable.

[0048] A program is understood in particular to be a computer program. A program is a sequence of instructions, in particular consisting of declarations and instructions, in order to be able to process and / or solve a specific functionality, task or problem on a digital data processing system. A program is usually in the form of software that is used with a digital data processing system. The program can in particular be in the form of firmware, in the case of the present invention in particular as firmware of the control device of the laboratory instrument. The program is usually available on a data carrier as an executable program file, frequently in so-called machine code, which is loaded into the main memory of the computer of the digital data processing system for execution. The program is processed as a sequence of machine, i.e. processor, instructions by the processor(s) of the computer and thus executed.The term 'computer program' also includes, in particular, the source code of the program from which the executable code can be created during the control of the laboratory device.

[0049] A control program is understood to be an executable computer program that preferably controls and / or carries out the desired treatment of the at least one sample, in particular depending on at least one program parameter. This program parameter can be a program parameter influenced and / or set by the user. The treatment can be controlled in particular by the control device generating one or more control parameters depending on the program parameters, by means of which control parameters the at least one treatment device is controlled. Preferably, the laboratory device has an operating system, which can be or can have a control program. The control program can in particular designate an operating system of the laboratory device or a component of the operating system. The operating system controls the treatment and other operating functions of the laboratory device.The control program can be determined by control parameters which can be derived by the control device from program parameters or user parameters.

[0050] The control program can, in particular, be signal-connected to the user interface device and / or can control the user interface device. The control device of the user interface device can be integrated into the control device of the laboratory device or can be designed separately from this control device. The control device of the user interface device can be integrated into the control system of the laboratory device, can be controllable by the control program, and / or can, in particular, be integrated into the control program. The control program can control further preferably provided functions of the laboratory device, for example an energy-saving function of the laboratory device or a communication function for communication with external data processing devices, which are, in particular, provided separately from the laboratory device and, in particular, are not a component of the laboratory device.

[0051] The thermoblock of the thermocycler, in particular, has a plurality of receptacles for sample containers. The control device of the thermocycler can be configured to acquire information in the form of sample container data associated with the runtime and the temperature control plan. For example, it is possible that the user performed a thermocyclically controlled reaction, in particular PCR, on an older TCx thermocycler while a certain number of sample containers of a certain type containing laboratory samples of a certain number and volume were arranged in the thermoblock of the TCx thermocycler. The evaluation program can be configured to consider such sample container data, in particular the number of sample containers, the type of sample container(s), and the number / volume of the laboratory samples, when determining the at least one temperature change rate from the runtime data and the temperature control plan data.

[0052] A sample container can be a single container containing only a single sample, or it can be a multiple container in which several individual containers are arranged connected to one another.

[0053] An individual container can be an open container or a sealable container. Sealable containers can include a lid element, particularly a closure cap. The lid element can be permanently attached to the container, e.g., as a hinged lid or a hinged closure cap, or it can be used as a separate component.

[0054] Preferably, in a multiple container, the multiple individual containers are arranged in fixed positions relative to one another, in particular according to the intersection points of a grid pattern. This simplifies the automated control of the positions and in particular the individual addressing of samples. A multiple container can be designed as a plate element in which the individual containers are connected such that they form a plate-shaped arrangement. The individual containers can be designed as depressions in a plate or can be connected to one another via web elements. The plate element can have a frame element in which the individual containers are held. These connections of components can be integral connections, i.e.The connections can be material-to-material connections and / or connections produced through a joint injection-molding process, or they can be force-fit and / or form-fit. The plate element can, in particular, be a microtiter plate.

[0055] Multi-well containers can comprise a plurality (from 2 to 10) of individual containers. They can also comprise a multiplicity (greater than 10), typically 12, 16, 24, 32, 48, 64, 96, 384, or 1536 individual containers. The multi-well container can, in particular, be a microtiter plate. A microtiter plate can be designed according to one or more industry standards, in particular the industry standards ANSI / SBS 1-2004, ANSI / SBS 2-2004, ANSI / SBS 3-2004, and ANSI / SBS 4-2004.

[0056] The maximum sample volume that can be accommodated by a sample container is typically between 0.01 ml and 100 ml, in particular 10 - 100 µl, 100 - 500 µl, 0.5 - 5 ml, 5-25 ml, 25-50 ml, 50-100 ml, depending on the type of transport container or sample vessel selected.

[0057] The sample container is preferably made partially or entirely of plastic. It is preferably a consumable item that is typically used for only one treatment or a small number of treatment steps of the sample. However, the sample container can also be made partially or entirely of another material.

[0058] Further preferred embodiments of the method according to the invention and of the thermocycler can be found in the description of the embodiments according to the figures.

[0059] They show: Fig. 1a shows the perspective front view of a thermal cycler according to the invention in one embodiment. Fig. 1b shows the perspective rear view of the thermal cycler of the Fig. 1a . Fig. 2a bis 2e show screen contents that appear on the screen of the thermocycler of the Fig. 1a und 1b can be displayed. Fig. 2e shows a screen input mask where the user, after entering the tempering plan, can enter the known runtime of the tempering plan, from which the thermocycler automatically calculates the temperature change rates. Fig. 3a shows an example of a user-defined temperature control plan, which in the thermocycler and method according to the invention is defined in particular by temperature control plan data. Fig. 3b shows schematically a temperature control plan, which is generated by the thermocycler and method according to the invention from the running time and the temperature plan data of the Fig. 3a is calculated. Fig . 3c shows schematically the temperature curve when changing between two temperature levels with the effective cooling rate marked as the difference quotient and the maximum cooling rate as the maximum differential. Fig. 4 shows schematically the sequence of an exemplary method according to the invention for determining at least one temperature change rate from temperature control plan data and runtime data Fig. 5 shows schematically the sequence of an exemplary method according to the invention for controlling a thermal cycler using the steps of the method for determining at least one temperature change rate from temperature control plan data and runtime data from Fig. 3 .

[0060] Fig. 1a shows a perspective front view of a thermal cycler 100 according to the invention in one exemplary embodiment. Externally, the thermal cycler 100 is characterized by a lid handle 1 for closing and opening the heated lid, the heated lid 2, and the approximately 10 cm thick, heat-insulating layer located in the heated lid to prevent condensation on the inside of the sample container.105°C heatable heating plate 3, the aluminum thermoblock 4 with 384 slots for PCR vessels, in particular a 384-well microtiter plate, which is contacted on its underside (not visible) with six Peltier elements, which form the temperature control elements of the thermocycler's temperature control device for heating and cooling the thermoblock and which are contacted on their underside (not visible) by a heat sink to dissipate the waste heat from the heat pumps to the environment, a power connection socket with power switch 5, a connection socket for Ethernet 6, a connection socket for data exchange with another thermocycler 7, a flap 8 to cover a USB port, a touchscreen 9 serving as a user interface device, and a nameplate 10. Alternatively, a 96-well aluminum or silver block can be used as an interchangeable thermoblock.

[0061] The thermal cycler 100 has a control device with a program-controlled microprocessor (not shown) which is designed to carry out the steps of the method 200 and 300 according to the invention in that the control program of the thermal cycler 100 is programmed to be able to carry out these steps.

[0062] The Fig. 3a shows a typical tempering plan that can be created by a user on the touchscreen 9 (see e.g. Fig. 2a ). It includes the desired (here: three) temperature levels 95°C, 65°C, 72°C and their holding times Δt1, Δt2, Δt3 of a cycle, which is to be repeated successively 30 times ("x30").

[0063] The Fig. 3b shows a typical temperature control plan, which is generated by the control unit of the thermocycler from the temperature plan data according to Fig. 3a and the runtime data entered by the user. The temperature control plan has the time intervals Δt S_cool , Δt S_heat1 and Δt S_heat2. In the time interval Δt S_cool the average temperature measured by temperature sensors of the temperature control device on the temperature block is cooled from 95°C to 65°C, for example at a cooling rate of 1.0°C / sec, which corresponds to the maximum cooling rate of a previously used, older thermal cycler TCx, which required the runtime now entered by the user to complete. Accordingly, in the time interval Δt S_heat1 the temperature is increased from 65°C to 72°C, for example at a heating rate of 2.0°C / sec, which corresponds to the maximum heating rate of the previously used, older thermal cycler TCx, which required the runtime now entered by the user to complete. In the time interval Δt S_heat2 the temperature is increased from 72°C to 95°C, e.g. with a heating rate of 2.0°C / sec, so that the cycle can start again.The thermal cycler according to the invention has a higher maximum heating and cooling rate, namely 10°C / sec and 5°C / sec, respectively, so that it can easily implement the calculated heating and cooling rates of the older devices. A settling time according to a standard settling control is also included in each time interval. As a result, the thermal cycler according to the invention simulates the temperature control behavior of the older device, allowing the user to easily reproduce their previously performed reaction protocols. This facilitates the migration from an older device to the thermal cycler according to the invention.

[0064] Figur 4 shows the exemplary method 200 according to the invention for determining at least one temperature change rate for controlling the temperature control device of a thermal cycler by calculating at least one temperature change rate from the known runtime of a known temperature control plan. The method 200 comprises the following steps: Providing temperature control plan data that determine the holding time and the temperature of at least one temperature level of the temperature control plan; (201) Providing runtime data that determine the runtime required to execute the temperature control plan on a thermal cycler; (202) Determining this at least one temperature change rate by an evaluation program using the temperature control plan data and the runtime data; (203) Providing the at least one previously determined temperature change rate for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate. (204)

[0065] Figur 5shows the exemplary method 300 according to the invention for controlling the temperature control device of a thermal cycler, wherein the thermal cycler comprises the temperature control device for controlling the temperature of a thermal block containing the samples for carrying out polymerase chain reactions in these samples according to the temperature control plan defined in the method according to one of claims 1 to 5, and an electronic control device configured to control the temperature control device using control parameters. The method 300 comprises the following steps: Providing temperature control plan data that determine the holding time and the temperature of at least one temperature level of the temperature control plan; (201) Providing runtime data that determine the runtime required to execute the temperature control plan on a thermal cycler; (202) Determining said at least one temperature change rate by an evaluation program using the temperature control plan data and the runtime data; (203) Providing the at least one previously determined temperature change rate for controlling the temperature control device of the thermal cycler as a function of said at least one temperature change rate.(204) Using the at least one previously determined temperature change rate to determine control parameters that include this at least one temperature change rate and that determine a temperature control plan corresponding to the temperature control plan; (301) Controlling the temperature control device by means of the control parameters and the electronic control device in order to execute this temperature control plan using this at least one temperature change rate. (302).

Claims

1. A method (200) for determining at least one temperature change rate for controlling the temperature control device of a thermal cycler, wherein the controller controls the temperature of a sample-receiving thermal block of the thermal cycler for performing polymerase chain reactions in said samples according to a temperature control schedule during which temperature stages are changed by changing the temperature at a temperature change rate, comprising the steps of: - Providing temperature control schedule data which determine the holding time and the temperature of at least one temperature stage of the temperature control schedule; (201) - Providing running time data that determines the running time required to execute the temperature control schedule on a thermal cycler, (202) - Determining of this at least one temperature change rate by an evaluation program using the temperature control schedule data and the running time data; (203) - Providing the at least one, previously determined, temperature change rate for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate. (204)2. The method according to claim 1, wherein the temperature control schedule data determine at least a first holding time and a first temperature of a first temperature stage and at least a second holding time and a second temperature of a second temperature stage of the temperature control schedule, wherein the first temperature is higher than the second temperature, wherein, according to the temperature control schedule, temperature stages are changed by cooling starting from the first temperature at a first temperature change rate (cooling rate) and heating starting from the second temperature at a second temperature change rate (heating rate). and wherein at least one first temperature change rate is determined by the evaluation program from the temperature control schedule data and the running time data, which is used as the cooling rate for setting the second temperature stage, and at least one second temperature change rate is determined, which is used as the heating rate for setting the first temperature stage, wherein these at least one cooling speed and at least one heating speed are provided for controlling the temperature control device of the thermal cycler.

3. The method according to claim 1 or 2, wherein during a cycle of the temperature control schedule there is at least one time interval during which at least one constant temperature change rate is applied and which includes a settling time period determined by the time period between the presence of the constant temperature change rate and the presence of a temperature stage to be set, during which time period a settling operation is carried out by the control of the temperature control device of the thermal cycler, which is part of the temperature control of a thermal cycler, comprising the step of: - Providing transient data containing information on at least one transient period, wherein the transient data is also used by the evaluation program when determining this at least one temperature change rate.

4. The method according to claim 3, comprising the step of: - Providing this transient data by input by a user at a user interface device of a data processing device by means of which the evaluation program is executed.

5. The method according to any one of the preceding claims, wherein the running time includes a latency interval during which, at the beginning of a temperature control schedule, a heatable lid covering the temperature control block of the thermal cycler containing the samples during the performance of the polymerase chain reaction is first set to a set temperature, wherein the running time data also include the information about this latency interval.

6. The method (300) for controlling the temperature control device of a thermal cycler, the method comprising the method according to any one of claims 1 to 5, which determines the at least one temperature change rate, wherein the thermal cycler comprises the temperature control device for controlling the temperature of a thermal block receiving the samples for performing polymerase chain reactions in said samples according to the temperature control schedule defined in the method according to any one of claims 1 to 5, and an electronic control device configured to control the temperature control device by means of control parameters, and the method comprises the steps of the method according to any one of claims 1 to 5 and the following steps: - Using the at least one previously determined at least one temperature change rate to determine control parameters which include said at least one temperature change rate and which determine a temperature control controlling schedule corresponding to the temperature control schedule; (301) - controlling the temperature control device by means of the control parameters and the electronic control device to execute said temperature control controlling schedule using said at least one temperature change rate. (302)7. A method for controlling the temperature control device of a first thermal cycler by simulating the temperature control behaviour of a second thermal cycler, the method comprising the method according to any one of claims 1 to 5, which determines the at least one temperature change rate characterizing the temperature control behaviour of the second thermal cycler, wherein the first thermal cycler is operable at a first maximum temperature change rate which is a cooling rate or a heating rate, and wherein the second thermal cycler is operable at a second maximum temperature change rate which is a cooling rate or a heating rate, wherein the first maximum temperature change rate is greater than or equal to the second maximum temperature change rate, wherein the first thermal cycler comprises the temperature control device for controlling the temperature of a thermal block receiving the samples for performing polymerase chain reactions in said samples according to the temperature control schedule defined in the method according to any one of claims 1 to 5, and an electronic control device adapted to control the temperature control device, and the method comprises the steps of the method according to any one of claims 1 to 5 and the following steps: - Using the at least one, previously determined, at least one temperature change rate to determine control parameters which include this at least one temperature change rate and which determine a temperature control controlling schedule corresponding to the temperature control schedule; - Controlling the temperature control device by means of the control parameters and the electronic control device to execute this temperature control controlling schedule using this at least one temperature change rate.

8. The method according to claim 7, wherein the at least one temperature change rate is less than the first maximum temperature change rate.

9. A thermal cycler (100), in particular for carrying out polymerase chain reactions in laboratory samples, comprising: - a temperature control device for controlling the temperature of a thermal block holding the samples according to a temperature control schedule, during which the temperature is changed between temperature stages by changing the temperature at the thermal block at a temperature change rate; - an electronic control device which comprises a data processing device and which is set up to control the temperature control device in order to carry out the following steps: • Using temperature control schedule data determining the temperature control schedule and running time data determining the running time of the temperature control schedule, and using the at least one temperature change rate previously determined according to the method according to any one of claims 1 to 5 to determine control parameters which include said at least one temperature change rate and which determine a temperature control controlling schedule corresponding to the temperature control schedule; • Controlling the temperature control device by means of the control parameters and the electronic control device to execute this temperature control controlling schedule using this at least one temperature change rate.

10. The thermal cycler according to claim 9, wherein the electronic control device is set up for carrying out the method according to one of claims 1 to 5, wherein the electronic control device is set up to execute an evaluation program by means of the data processing device of the electronic control device, and which is set up to carry out the following steps: - Recording of temperature control schedule data, which determine the holding time and the temperature of at least one temperature stage of the temperature control schedule; - Recording of running time data that determines the running time required to execute the temperature control schedule on a thermal cycler, - Determining of this at least one temperature change rate by means of the evaluation program using the temperature plan data and the running time data; - Use of the at least one temperature change rate determined by the evaluation program for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate.

11. The thermal cycler according to claim 9, wherein the data processing device of the electronic control device has an interface device via which a data connection can be established with an external data processing device, wherein on this external data processing device the method according to one of claims 1 to 5 has been carried out in order to provide the at least one temperature change rate, wherein the data processing device of the electronic control device is set up to receive this at least one temperature change rate, also the temperature control schedule data and / or the running time data, via the data connection.

12. The thermal cycler according to claim 9, comprising a user interface device, wherein the electronic control device is adapted to acquire the temperature control schedule data input by a user via the user interface device, and to acquire the running time data input by a user via the user interface device.

13. A program code which performs the following steps when executed by means of a data processing device of an electronic control device of a thermal cycler according to any one of claims 9 to 12: - Recording of temperature control schedule data, which are entered in particular by a user via a user interface device connected to the data processing device, which is a part of the thermal cycler, and which determine the holding time and the temperature of at least one temperature stage of the temperature control schedule; - Recording of running time data, which is entered in particular by a user via a user interface device connected to the data processing device, which is part of the thermal cycler, and which determines the running time used to execute the temperature control schedule on a thermal cycler, - Determining of this at least one temperature change rate by means of the evaluation program using the temperature control schedule data and the running time data, which is executed by the data processing device; - Providing of the at least one temperature change rate determined by the evaluation program for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate; - Using the at least one, previously determined, at least one temperature change rate to determine control parameters which include this at least one temperature change rate and which determine a temperature control controlling schedule; - Controlling the temperature control device by means of the control parameters and the electronic control device to execute this temperature control controlling schedule using this at least one temperature change rate.

14. Use of the method according to any one of claims 1 to 5 for controlling the temperature control device of a first thermal cycler by simulating the temperature control behavior of a second thermal cycler.