EP3626344A1 - Thermocycler and method for controlling a thermocycler - Google Patents

Abstract

The invention relates to a method for controlling a thermal cycler and a thermal cycler, in which at least one temperature change rate is determined by an evaluation program using temperature control data and runtime data, whereby in particular the temperature control behavior of a slower thermal cycler can be simulated on a faster thermal cycler.

Description

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, the control of which is set up to carry out such a method.

A thermal cycler is a laboratory device that is able to set the temperature of at least one laboratory sample in succession to a predetermined temperature and to keep it at this temperature level for a predetermined duration. The sequence of this temperature control is cyclical. This means that a predetermined temperature cycle, that is to say a sequence of at least two temperature levels, typically three temperature levels, is carried out repeatedly. This procedure is usually used to carry out a polymerase chain reaction (PCR).

When performing DNA duplication using a PCR, the user is dependent on the optimal result for the temperature being precisely and reproducibly tempered over the course of a temperature control schedule specified by the user.

The temperature cycle of a thermal cycler is specified by the user by setting a temperature control plan, usually directly on an operating device of the device. An example of such a cycle is the setting option in Fig. 2a shown with reference to a screen display of an exemplary thermal cycler according to the invention. The user sets the level of each temperature level of a cycle and its holding time, as well as the total number of cycles to be completed. The control of the thermal cycler ensures that the temperature control plan is implemented as precisely as possible according to these specifications. In most cases, the user cannot determine exactly, the device control uses certain control parameters that are adapted to the hardware components used in the thermal cycler. In particular, Peltier elements, which are used in the temperature control devices of the thermocyclers mostly as temperature control elements for temperature control of the sample block (thermoblocks) different performance values. The result of the tempering is also everyone

Hardware and material parameters of the thermal cycler 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.

Essential performance parameters of a thermal cycler are the maximum heating rate and the maximum cooling rate with which a thermal block of the thermal cycler can be tempered. Such a performance value is exemplary 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 characteristic of a thermal cycler or a class of thermal cyclers. The transient response is usually optimized to quickly reach a temperature level. The corresponding design of the temperature control is device-specific and is not known to the user. The manufacturers usually specify values for the maximum heating rate and the maximum cooling rate in the device specification. The actual temperature curve on the thermal block of the thermal cycler or the temperature curve in the liquid samples, which ultimately shapes the result of the temperature-dependent reaction sequences (e.g. PCR), depends not only on the user-defined temperature control schedule but also on the hardware-specific performance values mentioned.

In the studies on commercially available thermocyclers on which the invention is based, it has now been shown that the maximum temperature change rates or ramp rates are usually not reached or only for a short time. As in Figure 3c is shown, the knowledge of the maximum rate of temperature change, which is shown in the figure as the steepest straight line 2, does not directly _imply 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 area of the time interval is not equal to the difference quotient 1 in the time interval. The difference quotient in the time interval is subsequently also called effective temperature change rate or effective heating rate / effective cooling rate. The difference quotient, starting from a first point in time t1 (end of the point in time of the first temperature level) and a second point in time t2 (start of the point in time of the second temperature level) 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 given by the manufacturer is often only of limited significance and can even lead to incorrect conclusions with regard to the arithmetical estimate of the actual PCR runtime of a particular thermal cycler. In some cases, the runtimes of certain thermal cyclers were significantly longer than expected according to the ramp rates specified in the manufacturer's technical specifications. Slight deviations in the time until the completion of a PCR can generally occur due to environmental factors (e.g. room temperature, device placement, etc.). However, several reruns confirmed that such differences were only within a few seconds. It was concluded that the following aspects contributed greatly to the discrepancies observed: For the various thermal cyclers, the maximum ramp rates specified in the technical manuals are achieved for different periods during the ramp process from one temperature to the next - but may only be short for certain thermal cyclers Time during each ramp phase. Types of temperature control or the setting of the reaction volume can also significantly influence the ramp behavior. This can even result in a reaction having to be optimized again after the transfer of a PCR system from one thermal cycler to another.

The hardware-specific parameters are usually ignored by the user when carrying out a PCR. Instead, the temperatures are optimized (eg using a gradient) in order to obtain an optimal yield. When a PCR is migrated to another class of thermal cyclers, this yield usually decreases. This is not only due to the temperature deviating from the original device, but in particular also due to deviations in the dynamic behavior, ie on the ramp and during the transient response. Many users steer clear of that Repetition of attempts to switch to another device, as this does not produce the same result despite the same programming and application of the same temperature control plan. If the device is replaced, for example due to a defect or the device inventory is renewed, a complex requalification is required, especially when changing the device type, in order to reproduce earlier PCR results with the changed hardware. Since the operating programs of the thermal cycler offer only a limited possibility of influencing the control program and the control of the hardware components, a complex adaptation of the programming of the control may be necessary for the requalification.

The object on which the invention is based relates to the reproduction of a temperature profile in a thermal cycler when a user-defined temperature control plan is used again, in particular in a situation in which the hardware-specific performance values have changed, as is the case, for example, when the thermal cycler is changed.

In a preferred embodiment of the invention, the task is to be solved to make it easier for the user to switch from one thermal cycler to another thermal cycler with different performance values. For this purpose, the thermal cycler is designed in particular in accordance with the definitions of the present invention.

The invention solves this problem in each case by the method according to claim 1 and the thermal cycler according to claim 9 and by the program code according to claim 12. Preferred embodiments of the invention are the subject of the dependent claims and can also be found in the description and the figures of the invention.

The invention is based, among other things, on the observation that the entire runtime of the temperature control plan executed on the respective thermal cycler is easily ascertainable for the user and is generally also logged. The user pays attention to this runtime, since this is the essential parameter when developing the temperature control plan to determine the overall reaction time or to optimize the throughput is. According to the invention, the user specifies a known temperature control plan and a known runtime of this temperature control plan.

Applying the invention increases the probability of successfully reproducing a temperature profile. In particular, the dynamic behavior, including in particular the ramp rates and settling behavior, of an original thermal cycler is simulated or simulated without the user having to make great effort in the form of experiments and entering numerous parameters.

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\left(T_{s_{\mathit{heat}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{r_H}\right)+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}\left(T_{s_{\mathit{cool}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{cool}}}}{r_C}\right)$$

The temperature change rates, in particular the heating and cooling rates, which the temperature control device of the thermal cycler uses to reach the respective temperature level can be determined from the running time of a temperature control plan run through on a thermal cycler. The running time then includes the at least one holding time of the at least one temperature level of the temperature control plan and at least one time interval during which the setting of this at least one temperature level is carried out as a function of at least one temperature change rate. If one assumes, for example, 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 runtime T of the temperature control _plan consisting of repetitions of the same temperature _{cycle results} from the time intervals Δt _{S_heat} des Heating and the holding times T _{S_heat} at this higher temperature level and from the time intervals Δt _{S_cool} of cooling and the holding times T _{S_cool} at this low temperature level: $$T={\displaystyle \sum _{S_{\mathit{heat}}=1}^n}\left(T_{S_{\mathit{heat}}}+{\mathrm{\Delta }\mathit{t}}_{S_{\mathit{heat}}}\right)+{\displaystyle \sum _{S_{\mathit{cool}}=1}^m}\left(T_{S_{\mathit{cool}}}+{\mathrm{\Delta }\mathit{t}}_{S_{\mathit{cool}}}\right)$$

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\left(T_{s_{\mathit{heat}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{r_H}\right)+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}\left(T_{s_{\mathit{cool}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{cool}}}}{r_{\mathrm{C.}}}\right)$$

Here, r _{H is} the temperature change rate (heating rate) to heat the temperature control block by a temperature difference Δ _{S_heat} , and r _C is the temperature change rate (cooling rate) to cool the temperature control block by a temperature difference A _{S_cool} .

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\left(T_{s_{\mathit{heat}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{2}\frac{1}{r_C}\right)+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}\left(T_{s_{\mathit{cool}}}+{\mathrm{\Delta }}_{s_{\mathit{cool}}}\frac{1}{r_C}\right)$$

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}+\frac{1}{r_C}{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{2}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}+\frac{1}{r_C}{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}$$

$$\begin{array}{c}T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}+\frac{1}{r_C}\left({\displaystyle \sum _{s_{\mathit{hea}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{hea}}}}{2}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}\right)\\ T-{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}-{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}=\frac{1}{r_C}\left({\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{2}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}\right)\end{array}$$

$$\boxed{r_c=\frac{\sum _{s_{\mathit{heat}}=1}^n\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{2}+\sum _{s_{\mathit{cool}}=1}^m{\mathrm{\Delta }}_{s_{\mathit{cool}}}}{T-\sum _{s_{\mathit{heat}}=1}^n{T}_{s_{\mathit{heat}}}-\sum _{s_{\mathit{cool}}=1}^m{T}_{s_{\mathit{cool}}}},r_H=2r_C},$$

unter der Bedingung, dass $$T>{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}-{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}$$

In the context of the present application, unless otherwise described, the terms heating rate, cooling rate and rate of temperature change respectively denote the effective heating rate, the effective cooling rate and the effective rate of temperature change, and not, for example, extreme values which are present for a short time when the temperature changes. In most commercially available thermal cyclers, the effective heating rate and the effective cooling rate are roughly constant. This can be easily determined for known thermal cyclers and saved 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 ratio can be specified with the r _H = approach with sufficient accuracy 2 * r _C in many cases thermocycler. Then the effective cooling rate or effective heating rate can be calculated from the running time as follows: $$T={\displaystyle \sum _{s_{\mathit{hea}}=1}^n}\left(T_{s_{\mathit{heat}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{\mathrm{2nd}{r}_{\mathrm{C.}}}\right)+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}\left(T_{s_{\mathit{cool}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{cool}}}}{r_{\mathrm{C.}}}\right)$$

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\left(T_{s_{\mathit{heat}}}+\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{\mathrm{2nd}}\frac{1}{r_{\mathrm{C.}}}\right)+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}\left(T_{s_{\mathit{cool}}}+{\mathrm{\Delta }}_{s_{\mathit{cool}}}\frac{1}{r_{\mathrm{C.}}}\right)$$

$$T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}+\frac{1}{r_{\mathrm{C.}}}{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{\mathrm{2nd}}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}+\frac{1}{r_{\mathrm{C.}}}{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}$$

$$\begin{array}{c}T={\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}+\frac{1}{r_{\mathrm{C.}}}\left({\displaystyle \sum _{s_{\mathit{hea}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{hea}}}}{\mathrm{2nd}}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}\right)\\ T-{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}-{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}=\frac{1}{r_{\mathrm{C.}}}\left({\displaystyle \sum _{s_{\mathit{heat}}=1}^n}\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{\mathrm{2nd}}+{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{\mathrm{\Delta }}_{s_{\mathit{cool}}}\right)\end{array}$$

$$\boxed{r_c=\frac{\sum _{s_{\mathit{heat}}=1}^n\frac{{\mathrm{\Delta }}_{s_{\mathit{heat}}}}{\mathrm{2nd}}+\sum _{s_{\mathit{cool}}=1}^m{\mathrm{\Delta }}_{s_{\mathit{cool}}}}{T-\sum _{s_{\mathit{heat}}=1}^n{T}_{s_{\mathit{heat}}}-\sum _{s_{\mathit{cool}}=1}^m{T}_{s_{\mathit{cool}}}},r_H=\mathrm{2nd}{r}_{\mathrm{C.}}},$$

under the condition that $$T>{\displaystyle \sum _{s_{\mathit{heat}}=1}^n}{T}_{s_{\mathit{heat}}}-{\displaystyle \sum _{s_{\mathit{cool}}=1}^m}{T}_{s_{\mathit{cool}}}$$

This or a calculation that is comparable in the 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 carried out on a computer or on the data processing device of a thermal cycler according to the invention. The rates of temperature change are determined in particular on the assumption of a medium transient response (standard). This is exemplary 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 up and cooling down each contain a constant time period (settling time period), which takes into account the settling of the control loop to the respective level of the temperature level. The settling time period is to be set by the period between the presence of the constant temperature change rate and the presence of a Temperature level determined. Alternatively, the settling time period for certain commercial thermal cyclers can also be determined and saved in a table. In addition, a table can contain a pre-selection 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. As an alternative or in addition, the thermal cycler or method according to the invention can be set up so that the user changes the constant value of the settling time period as a variable, in particular by input via a user interface device of the thermal cycler and a data input in this way. The user can thus easily correct the result by changing the transient response if the yield is not correct in the method.

A temperature control of the temperature control block of a thermal cycler that is optimized for the controlled settling is known in principle. In the case of heating, the oscillation to the desired target temperature of a temperature level of the temperature cycle uses an overshoot of the temperature set in the temperature control block to a maximum temperature value which is above the setpoint to be set, followed by an undershoot to a temperature value below the setpoint, to then switch back to a lower temperature value above the setpoint, etc., until the setpoint is reached. The settling can then be characterized by the temperature difference between the maximum overshoot temperature (for cooling: the minimum undershoot temperature) and the duration of the overshoot until the target temperature is reached. With a rapid settling, this temperature difference and the duration are small. In particular, the user can be given a preselection from a limited number of transient modes for selection. Each such settling mode can be characterized by specific values for the temperature difference and duration mentioned, each 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 list selection on the display, for example under the designation “Fast”, “Intermediate”, “Standard”, “Safe”, as is the case in the exemplary embodiment of Figure 2d is provided. Details on the implementation of such temperature controls using "overshoot" are known, for example, from EP 0 488 769 A2 , described there as a "controlled overshoot algorithm". The temperature control can also be carried out by additionally observing the effect of the temperature control on the temperature of the liquid sample contained in the vessel which is inserted into the thermal block of the thermal cycler, as is known, for example, from EP 1 452 608 B1 .

The method according to the invention and / or the thermal cycler according to the invention can be set up in such a way that a particular commercial thermal cycler TC _X can be selected by the user, in particular via a user interface device of the thermal cycler, for example via a list selection that is displayed on a display or touchscreen of the user interface device and can be operated. The thermal cycler or the method then has the additional information that the runtime T specified by the user relates to the implementation of the temperature control plan on the thermal cycler of the TC _X type. 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 rates of temperature change can be made according to the selection made by the user TC _X done automatically.

• Bereitstellen von Temperierplandaten, die die Haltezeit und die Temperatur mindestens einer Temperaturstufe des Temperierplans bestimmen;
• Bereitstellen von Laufzeitdaten, welche die Laufzeit bestimmen, die zur Ausführung des Temperierplans auf einem Thermocycler benötigt wird,
• Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit durch ein die Temperieplandaten und die Laufzeitdaten verwendendes Auswertungsprogramm;
• Bereitstellen der mindestens einen, zuvor ermittelten, Temperaturänderungsgeschwindigkeit für die Steuerung der Temperiervorrichtung des Thermocyclers in Abhängigkeit von dieser mindestens einen Temperaturänderungsgeschwindigkeit.

According to the invention, the method is used to determine at least one temperature change rate for the control of the temperature control device of a thermal cycler, the control tempering a sample-receiving thermoblock of the thermal cycler for carrying out polymerase chain reactions in these samples according to a temperature control schedule, during which the temperature is changed by changing the temperature with a Temperature change rate is changed, comprising the steps:

• Providing temperature control plan data which determine the holding time and the temperature of at least one temperature level of the temperature control plan;
• Provision of runtime data, which determine the runtime required to execute the temperature control plan on a thermal cycler,
• Determining this at least one rate of temperature change by means of an evaluation program using the temperature plan data and the runtime data;
• Provision of 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.

The temperature plan data preferably determine at least a first hold time and a first temperature of a first temperature level and at least a second hold time and a second temperature of a second temperature level of the temperature plan. Typically, a cycle of the temperature control plan also has three temperature levels, if it is a PCR, so that the temperature control plan data also determine a third holding time and a third temperature of the third temperature level. The first temperature is assumed to be higher than the second temperature, and according to the temperature control plan, switching between temperature levels is carried out by cooling from the first temperature with a first temperature change rate (cooling rate) and heating starting from the second temperature with a second temperature change rate (heating rate) becomes.

The evaluation program preferably determines the 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 rate of change which is used as the heating rate for setting the first temperature level, is determined from the temperature control plan data and the runtime data. These at least one cooling rate and at least one heating rate are preferably provided for the control of the temperature control device of the thermal cycler.

• Bereitstellen von Einschwingdaten, die Informationen über mindestens eine Einschwingzeitperiode enthalten,

wobei bei der Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit durch das Auswertungsprogramms insbesondere auch die Einschwingdaten verwendet werden.During a cycle of the temperature control schedule, in particular there is at least one time interval during which at least one constant rate of temperature change is used, and that also one Settling time period, which can be determined by the period between the presence of the constant rate of temperature change and the presence of a temperature level to be set, during which 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, which The method comprises the step:

• Providing settling data that contains information about at least one settling period,

the transient data in particular also being used in the determination of this at least one temperature change rate by the evaluation program.

• Bereitstellen dieser Einschwingdaten durch Eingabe eines Benutzers an einer Benutzerschnittstelleneinrichtung einer Datenverarbeitungseinrichung, mittels der das Auswertungsprogramm ausgeführt wird.

The method preferably includes the step:

• Providing this transient data by inputting a user at a user interface device of a data processing device, by means of which the evaluation program is executed.

The runtime can also include a latency interval, during which, at the beginning of a temperature control plan, a heatable cover, which covers the temperature control block of the thermal cycler containing the samples during the implementation of the polymerase chain reaction, is set to a target temperature, the runtime data also providing information about this latency interval include.

• Verwenden der mindestens einen, zuvor ermittelten, mindestens einen Temperaturänderungsgeschwindigkeit zur Bestimmung von Steuerungsparametern, welche diese mindestens eine Temperaturänderungsgeschwindigkeit beinhalten und welche einen dem Temperierplan entsprechenden Temperiersteuerungsplan bestimmen;
• Steuern der Temperiervorrichtung mittels der Steuerungsparameter und der elektronischen Steuerungseinrichtung, um diesen Temperiersteuerungsplan unter Verwendung dieser mindestens einen Temperaturänderungsgeschwindigkeit auszuführen.

The method according to the invention is preferably used to control the temperature control device of a thermal cycler, the method of controlling the temperature control device preferably containing a method for determining at least one temperature change rate from runtime data and temperature control plan data. The thermal cycler has the temperature control device for temperature control of a thermoblock holding the samples for carrying out Polymerase chain fractions in these samples in accordance with the temperature control plan mentioned in the method according to the invention, and has an electronic control device which is set up to control the temperature control device by means of control parameters. The method of controlling the temperature control device of a thermal cycler has the steps of the method for determining at least one temperature change rate from runtime data and temperature control plan data and the following steps:

• Using the at least one, previously determined, at least one temperature change speed for determining control parameters which contain these at least one temperature change speed 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 execute this temperature control plan using this at least one temperature change rate.

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, the method for controlling the temperature control device including the method for determining at least one temperature change rate from temperature control plan data and runtime data, which control the temperature control behavior of the second thermal cycler characterize. In particular, the first thermal cycler can be operated at a first maximum temperature change rate, which is a cooling rate or a heating rate, and the second thermal cycler can in particular be operated at a second maximum temperature change rate, which is a cooling rate or a heating rate, the first maximum temperature rate of change being greater or is equal to the second maximum rate of temperature change. In short, the first thermal cycler preferably tempers faster than the second thermal cycler. Example of such a thermal cycler, whose maximum effective heating and cooling rate is greater than most The maximum effective heating and cooling rates of other commercially available thermocyclers 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.

• Verwenden der mindestens einen, zuvor ermittelten, mindestens einen Temperaturänderungsgeschwindigkeit zur Bestimmung von Steuerungsparametern, welche diese mindestens eine Temperaturänderungsgeschwindigkeit beinhalten und welche einen dem Temperierplan entsprechenden Temperiersteuerungsplan bestimmen;
• Steuern der Temperiervorrichtung mittels der Steuerungsparameter und der elektronischen Steuerungseinrichtung, um diesen Temperiersteuerungsplan un-ter Verwendung dieser mindestens einen Temperaturänderungsgeschwindigkeit auszuführen.

The first thermal cycler has the temperature control device for temperature control of a thermoblock holding the samples for carrying out polymerase chain reactions in these samples in accordance with the temperature control plan defined in the method for determining at least one temperature change rate, and an electronic control device which is set up to control the temperature control device. The method for controlling the temperature control device has, 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, at least one temperature change speed for determining control parameters which contain these at least one temperature change speed 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 execute this temperature control plan using this at least one temperature change rate.

The at least one rate of temperature change is in particular smaller than the first maximum rate of temperature change.

• eine Temperiervorrichtung zum Temperieren eines die Proben aufnehmenden Thermoblocks gemäß eines Temperierplans, während dem zwischen Temperaturstufen gewechselt wird, indem die Temperatur am Thermoblock mit einer Temperaturänderungsgeschwindigkeit geändert wird;
• eine elektronische Steuerungseinrichtung, die eine Datenverarbeitungseinrichtung aufweist und die zur Steuerung der Temperiervorrichtung eingerichtet ist, um die folgenden Schritte durchzuführen:
  • Verwenden von Temperierplandaten, die den Temperierplan bestimmen, und von Laufzeitdaten, die die Laufzeit des Temperierplans bestimmen, und Verwenden der mindestens einen, zuvor, insbesondere gemäß des Verfahrens zur Ermittlung mindestens einer Temperaturänderungsgeschwindigkeit aus Temperierplandaten und Laufzeitdaten, ermittelten, mindestens einen Temperaturänderungsgeschwindigkeit zur Bestimmung von Steuerungsparametern, welche diese mindestens eine Temperaturänderungsgeschwindigkeit beinhalten und welche einen dem Temperierplan entsprechenden Temperiersteuerungsplan bestimmen;
  • Steuern der Temperiervorrichtung mittels der Steuerungsparameter und der elektronischen Steuerungseinrichtung, um diesen Temperiersteuerungsplan unter Verwendung dieser mindestens einen Temperaturänderungsgeschwindigkeit auszuführen.

The invention relates to a thermal cycler, in particular for carrying out polymerase chain fractions in laboratory samples, comprising:

• a tempering device for tempering a thermoblock receiving the samples according to a tempering plan during which switching between temperature levels by changing the temperature at the thermoblock with a temperature change rate;
• an electronic control device which has 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 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 speed for determining at least one temperature previously determined, in particular according to the method for determining at least one temperature change speed from temperature control plan data and runtime data Control parameters which contain 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 execute this temperature control plan using this at least one temperature change rate.

• Erfassen von Temperierplandaten, welche die die Haltezeit und die Temperatur mindestens einer Temperaturstufe des Temperierplans bestimmen;
• Erfassen von Laufzeitdaten, welche die Laufzeit bestimmen, die zur Ausführung des Temperierplans auf einem Thermocycler benötigt wird,
• Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit mittels des die Temperieplandaten und die Laufzeitdaten verwendenden Auswertungsprogramms;
• Verwenden der mindestens einen, durch das Auswertungsprogramm ermittelten Temperaturänderungsgeschwindigkeit für die Steuerung der Temperiervorrichtung des Thermocyclers in Abhängigkeit von dieser mindestens einen Temperaturänderungsgeschwindigkeit.

The electronic control device of the thermal cycler is set up, in particular, to carry out the method for determining at least one temperature change rate from temperature control plan data and runtime data, the electronic control device being 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:

• Acquisition of tempering plan data which determine the holding time and the temperature of at least one temperature level of the tempering plan;
• Acquisition of runtime data that determine the runtime required to execute the temperature control plan on a thermal cycler,
• Determining this at least one rate of temperature change 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.

The data processing device of the electronic control device preferably has an interface device via which a data connection to an external data processing device can be established, the method for determining at least one temperature change rate from temperature control plan data and runtime data being carried out in particular 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 set up to receive this at least one temperature change rate, in particular also the temperature control plan data and / or the runtime data, via the data connection.

The thermal cycler preferably has a user interface device, the electronic control device being set up 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.

• Erfassen von Temperierplandaten, die insbesondere von einem Benutzer über eine mit der Datenverarbeitungseinrichtung verbundenen Benutzerschnittstelleneinrichtung, die insbesondere ein Teil des Thermocyclers ist, eingegeben werden und welche die die Haltezeit und die Temperatur mindestens einer Temperaturstufe des Temperierplans bestimmen;
• Erfassen von Laufzeitdaten, die insbesondere von einem Benutzer über eine mit der Datenverarbeitungseinrichtung verbundenen Benutzerschnittstelleneinrichtung, die insbesondere ein Teil des Thermocyclers ist, eingegeben werden und welche die Laufzeit bestimmen, die zur Ausführung des Temperierplans auf einem Thermocycler verwendet wird,
• Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit mittels des die Temperierplandaten und die Laufzeitdaten verwendenden Auswertungsprogramms, das insbesondere von der Datenverarbeitungseinrichtung ausgeführt wird;
• Bereitstellen der mindestens einen, durch das Auswertungsprogramm ermittelten Temperaturänderungsgeschwindigkeit für die Steuerung der Temperiervorrichtung des Thermocyclers in Abhängigkeit von dieser mindestens einen Temperaturänderungsgeschwindigkeit;
• Verwenden der mindestens einen, zuvor ermittelten, mindestens einen Temperaturänderungsgeschwindigkeit zur Bestimmung von Steuerungsparametern, welche diese mindestens eine Temperaturänderungsgeschwindigkeit beinhalten und welche einen Temperiersteuerungsplan bestimmen;
• Steuern der Temperiervorrichtung mittels der Steuerungsparameter und der elektronischen Steuerungseinrichtung, um diesen Temperiersteuerungsplan unter Verwendung dieser mindestens einen Temperaturänderungsgeschwindigkeit auszuführen.

The invention also relates to a program code which carries out the following steps when it is executed by means of a data processing device, in particular the data processing device of an electronic control device of a thermal cycler:

• Acquisition of temperature control plan data which are entered in particular by a user via a user interface device which is connected to the data processing device and which is in particular part of the thermal cycler and which determine the holding time and the temperature of at least one temperature level of the temperature control plan;
• Acquisition of runtime data, which are entered in particular by a user via a user interface device which is connected to the data processing device and is in particular part of the thermal cycler and which determine the runtime which is used to execute the temperature control plan on a thermal cycler.
• Determining this at least one temperature change rate by means of the evaluation program using the temperature control plan data and the runtime data, which is executed in particular 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, at least one temperature change rate for determining control parameters which contain these at least one temperature change rate and which 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.

The invention also surpasses 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 helps the user migrate from the older, less powerful second thermal cycler to the more powerful first thermal cycler.

A thermal cycler is a device that is able to set the temperature of at least one sample in succession to a predetermined temperature and to keep it at this temperature level for a predetermined holding time. The sequence of this temperature control is cyclical. That is, a predetermined temperature cycle, that is a sequence of at least two temperature levels is carried out repeatedly. This method is used in particular to carry out a polymerase chain reaction (PCR).

A thermal cycler, in particular the treatment device of the thermal cycler, preferably has a thermal block. A thermoblock is a sample holder made of a heat-conducting material, usually a metal-containing material or a metal, especially aluminum or silver. The sample holder has a contacting 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.

The thermal cycler, in particular the treatment device of the thermal cycler, has a control device with at least one control circuit, to which the at least one heating / cooling device is assigned as an actuator and at least one temperature measuring device as a measuring element. The temperature of a temperature level is regulated by means of the control device. A heat sink of the thermal cycler is used to cool sections of the thermal cycler, in particular the cooling of the Peltier elements.

The thermal cycler, in particular the treatment device of the thermal cycler, can have further heating and / or cooling elements. The thermal cycler, in particular the treatment device of the thermal cycler, preferably has a timer device with which time parameters for setting the temperature cycle can be controlled. In the case of 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 Process parameter that influences or defines the process, in particular the sequence, of a temperature control program consisting of several steps.

The thermal cycler has, in particular, an electronic control device. Within the scope of the present invention, a control device generally has, in particular, a data processing device, in particular a computing unit (CPU) for processing data and / or a microprocessor, or is a data processing device. The control device or a computing unit of the control device of a thermal cycler is preferably also set up for program-based control of the temperature control of the thermoblock.

The data processing device preferably has a computing unit, in particular a CPU, further preferably at least one data storage device, in particular for the 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.

The use of a thermal cycler for the cyclical temperature control of laboratory samples, in particular for carrying out a PCR in these laboratory samples, is a device-controlled treatment, that is to say in particular an at least partially automated treatment. In the case of a partially automated treatment, it is in particular possible for the treatment to be carried out in such a way that at least one user input is made after the start of the treatment and before the end of the treatment, with which the user can influence the ongoing treatment, in particular, for example, by using one User interface device of the thermal cycler answers automatic query, in particular confirms or denies an entry or makes other entries. In the case of the partially automated treatment, it is possible in particular for the treatment to have a plurality of treatment steps, which are in particular carried out automatically one after the other in time, and to have at least one treatment step which requires user input, in particular via a user interface device. examples for Such user inputs on a thermal cycler are here the input of temperature control data, the input of the runtime, and / or optionally the input or selection of a thermal cycler TC _X assigned to the input runtime.

The device-controlled treatment is preferably a program-controlled treatment, that is to say a treatment controlled by a program. A program-controlled treatment of a sample is understood to mean that the treatment process essentially takes place by processing a plurality or a plurality of program steps. The program-controlled treatment is preferably 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 for a thermal cycler determine the temperature control plan, in particular the height and holding time of the temperature levels of a temperature control cycle, the total number of cycles, and in the context of the present invention also the runtime of the temperature control plan, which is known to the user from previous thermal cycler applications of the same temperature control plan. The program-controlled treatment is preferably carried out with the aid of a digital data processing device, which can in particular be part of the control device of the laboratory device. The data processing device can have at least one processor, i.e. have a CPU and / or have at least one microprocessor. The program-controlled treatment is preferably controlled and / or carried out in accordance with the specifications of a program, in particular a control program. In particular, in the case of a program-controlled treatment, at least after the program parameters required by the user have been recorded, essentially no user activity is required.

A program parameter is understood to be a variable that can be set in a predetermined manner within a program or subroutine, valid for at least one execution (call) of the program or subroutine. The program parameter is determined, for example by the user, and controls the program or subroutine and effects data output depending on this program parameter. In particular, the program parameter influences and / or controls and / or the data output by the program control the control of the device, in particular the control of the treatment by means of the at least one treatment device.

A program parameter can be a program parameter required by the user. A program parameter required by the user is characterized in that it is required for the execution of 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 otherwise available, in particular optionally set by the user. A program parameter is set by a user in particular by displaying a selection of possible predefined values from a list of predefined values stored in the laboratory device, the user selecting and thus setting the desired parameter from this list. This applies, for example, to the selection of a TCx thermal cycler, which the user assigns to a known runtime of a temperature control plan. It is also possible that this program parameter is set by the user entering the value, e.g. Use a numeric keypad to enter a number that corresponds to the desired value or by the user increasing or decreasing a value continuously or in increments until it corresponds to the desired value and thus setting the value. Other forms of input, e.g. voice control and / or gesture control are conceivable.

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 available as software that is used with a digital data processing system. The program can be present in particular as firmware, in the case of the present invention in particular as firmware of the control device of the laboratory device. The program is usually on a data carrier as an executable program file, often in the so-called machine code, which is loaded into the working memory of the computer of the digital data processing system for execution. The program is processed and executed by the processor (s) of the computer as a sequence of machine, ie processor instructions. 'Computer program' is also understood to mean, in particular, the source text of the program, from which the executable code can be generated in the course of the control of the laboratory device.

A control program is understood to be an executable computer program which preferably controls and / or carries out the desired treatment of the at least one sample, in particular as a function of at least one program parameter. This program parameter can be a program parameter influenced and / or set by the user. The treatment can in particular be controlled by the control device generating one or more control parameters as a function of the program parameters, by means of which the at least one treatment device is controlled. The laboratory device preferably 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 determinable by control parameters which can be derived from the control device from program parameters or user parameters.

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 in the control device of the laboratory device or can be formed separately from this control device. The control device of the user interface device can be integrated in the control of the laboratory device, can be controllable by the control program and / or can in particular be integrated in the control program. The control program can control further functions of the laboratory device that are preferably provided, 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 part of the laboratory device.

The thermal block of the thermal cycler has in particular a large number of receptacles for sample containers. The control device of the thermal cycler can be set up to acquire information in the form of sample container data which are associated with the running time and the temperature control schedule. It is possible, for example, for the user to carry out a thermocyclically controlled reaction, in particular PCR, on an older thermocycler TCx, while a certain number of sample containers of a certain type with laboratory samples of a certain number and a certain volume were arranged in the thermoblock of the thermocycler TCx. The evaluation program can be set up to take such sample container data, in particular the number of sample containers, the type of sample container (s), number / volume of laboratory samples, into account when determining the at least one rate of temperature change from the runtime data and the temperature control plan data.

A sample container can be a single container in which only a single sample is contained, or can be a multiple container in which a plurality of individual containers are arranged connected to one another.

A single container can be an open container or a closable container. In the case of a closable container, a cover element, in particular a closure cap, can be provided. The lid element can be firmly connected to the container, e.g. as a hinged lid or hinged closure cap, or can be used as a separate component.

In the case of a multiple container, the plurality of individual containers are preferably arranged in fixed positions relative to one another, in particular arranged according to the crossing points of a grid pattern. This simplifies the automated control of the positions and in particular the individual addressing of samples. A multiple relationship can be designed as a plate element in which the individual containers are connected in such a way that they form a plate-shaped arrangement. The individual containers can or can be designed as depressions in a plate 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, that is to say integral connections and / or connections produced by a common injection molding process, or can be produced with a force fit (English "force fit") and / or form fit (English "form fit"). The plate element can in particular be a microtiter plate.

Multiple containers can have a plurality (from 2 to 10) of individual containers. They can also have a large number (greater than 10), typically 12, 16, 24, 32, 48, 64, 96, 384, 1536 individual containers. The multiple 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, ANSI / SBS 4-2004.

The maximum sample volume that can be taken up by a sample ratio 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 container selected.

The sample container is preferably partially or completely made of plastic. It is preferably a consumable that is typically used only for one treatment or a small number of treatment steps of the sample. However, the sample container can also consist partially or completely of another material.

Preferred embodiments of the thermal cycler according to the invention can be found in particular in the description of one of the methods according to the invention. Preferred configurations of the processes according to the invention can be found in particular in the description of the thermal cyclers according to the invention.

Further preferred configurations of the method according to the invention and of the thermal cycler can be found in the description of the exemplary embodiments according to the figures.

• Fig. 1a zeigt die perspektivische Vorderansicht eines Thermocyclers gemäß der Erfindung in einem Ausführungsbeispiel.
• Fig. 1b zeigt die perspektivische Rückansicht des Thermocyclers der Fig. 1a.
• Fig. 2a bis 2e zeigen jeweils Bildschirminhalte, die auf dem Bildschirm des Thermocyclers der Fig. 1a und 1b angezeigt werden können.
• Fig. 2e zeigt eine Bildschirmeingabemaske, bei der der Benutzer nach Eingabe des Temperierplans die ihm bekannte Laufzeit (runtime) des Temperierplans eingeben kann, aus der der Thermocycler selbständig die Temperaturänderungsgeschwindigkeiten berechnet.
• Fig. 3a zeigt exemplarisch einen vom Benutzer definierten Temperierplan, der beim erfindungsgemäßen Thermocycler und Verfahren insbesondere durch Temperierplandaten definiert ist.
• Fig. 3b zeigt schematisch einen Temperiersteuerungsplan, der vom erfindungsgemäßen Thermocycler und Verfahren aus der Laufzeit und den Temperierplandaten der Fig. 3a berechnet wird.
• Fig . 3c zeigt schematisch den Temperaturverlauf beim Wechsel zwischen zwei Temperaturstufen mit der Kennzeichnung der effektiven Kühlrate als Differenzenquotienten und der maximalen Kühlrate als maximales Differential.
• Fig. 4 zeigt schematisch den Ablauf eines beispielhaften erfindungsgemäßen Verfahrens zur Ermittlung mindestens einer Temperaturänderungsgeschwindigkeit aus Temperierplandaten und Laufzeitdaten
• Fig. 5 zeigt schematisch den Ablauf eines beispielhaften erfindungsgemäßen Verfahrens zur Steuerung eines Thermocyclers unter Verwendung der Schritte des Verfahrens zur Ermittlung mindestens einer Temperaturänderungsgeschwindigkeit aus Temperierplandaten und Laufzeitdaten aus Fig. 3.

Show it:

• Fig. 1a shows the perspective front view of a thermal cycler according to the invention in one embodiment.
• Fig. 1b shows the rear perspective view of the thermal cycler of Fig. 1a .
• 2a to 2e each show the screen contents on the screen of the thermal cycler of the 1a and 1b can be displayed.
• Fig. 2e shows a screen input mask in which the user can enter the known runtime of the temperature control plan after entering the temperature control plan, from which the thermal cycler independently calculates the temperature change rates.
• Fig. 3a shows an example of a temperature control plan defined by the user, which is defined in the thermal cycler and method according to the invention in particular by temperature control data.
• Fig. 3b shows schematically a temperature control plan, the thermal cycler and method according to the invention from the runtime and the temperature plan data of Fig. 3a is calculated.
• Fig. 3c shows schematically the temperature curve when changing between two temperature levels with the identification of the effective cooling rate as the difference quotient and the maximum cooling rate as the maximum differential.
• Fig. 4 schematically shows 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 schematically shows 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 plan data and runtime data Fig. 3 .

Fig. 1a shows the perspective front view of a thermal cycler 100 according to the invention in one embodiment. Externally, the thermal cycler 100 is characterized by a lid handle 1 for closing and opening the heated lid, the heated lid 2, the heated plate 3 located in the heated lid to prevent condensation on the inside of the sample container with a heating plate 3 that can be heated to about 105 ° C., the aluminum thermoblock 4 with here 384 recordings for holding PCR tubes, in particular a 384 microtiter plate, which is contacted on its underside (not visible) here with six Peltier elements, which form the temperature control elements of the temperature control device of the thermal cycler for heating and cooling the thermoblock, and those on the underside ( are not visible) are contacted by a heat sink in order to dissipate the waste heat from the heat pumps to the environment, a mains connection socket with mains switch 5, a connection socket for Ethernet 6, a connection socket for data exchange with another thermal cycler 7, a flap 8 for covering a USB Connection, one as a user interface elleneinrichtung serving touchscreen 9, a nameplate 10. Alternatively, a 96-series aluminum or silver block can be used as an interchangeable thermoblock.

The thermal cycler 100 has a control device with a program-controlled microprocessor (not shown), which is set up 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.

The Fig. 3a shows a typical temperature control plan, which a user on the touch screen 9 (see, for example Fig. 2a ) may have been defined. It includes the desired (here: three) temperature levels 95 ° C, 65 ° C, 72 ° C and their holding times Δt1, Δt2, Δt3 of a cycle that is to be repeated successively 30 times ("x30").

The Fig. 3b shows a typical temperature control plan, which according to the control device of the thermal cycler from the temperature plan data Fig. 3a and was calculated based on the runtime data entered by the user. The temperature control _schedule has the time intervals At _{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 with the cooling rate 1.0 ° C / sec, which corresponds to the maximum cooling rate of an older thermocycler TCx used previously , which had required the runtime now entered by the user. Correspondingly, the temperature is increased from 65 ° C to 72 ° C in the time interval Δt _{S_heat1} , for example with a heating rate of 2.0 ° C / sec, which corresponds to the maximum heating rate of the older thermocycler TCx used previously, which is carried out by the user had now entered entered runtime. In the time interval Δt _{S_heat2} the temperature is increased from 72 ° C to 95 ° C, for example 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 larger maximum heating and cooling rate, namely 10 ° C./sec and 5 ° C./sec, so that it can easily carry out the calculated heating and cooling rates of the older devices. A settling time according to a standard settling control is also included in the time interval. As a result, the thermal cycler according to the invention simulates the temperature control behavior of the older device, so that the user can easily reproduce his previously performed reaction protocols. In this way, the migration from an older device to the thermal cycler according to the invention is facilitated.

• Bereitstellen von Temperierplandaten, die die Haltezeit und die Temperatur mindestens einer Temperaturstufe des Temperierplans bestimmen; (201)
• Bereitstellen von Laufzeitdaten, welche die Laufzeit bestimmen, die zur Ausführung des Temperierplans auf einem Thermocycler benötigt wird, (202)
• Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit durch ein die Temperieplandaten und die Laufzeitdaten verwendendes Auswertungsprogramm; (203)
• Bereitstellen der mindestens einen, zuvor ermittelten, Temperaturänderungsgeschwindigkeit für die Steuerung der Temperiervorrichtung des Thermocyclers in Abhängigkeit von dieser mindestens einen Temperaturänderungsgeschwindigkeit. (204)

Figure 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 Velocity of temperature change from the known running time of a known temperature control plan. The method 200 here comprises the following steps:

• Providing temperature control plan data which determine the holding time and the temperature of at least one temperature level of the temperature control plan; (201)
• Provision of runtime data which determine the runtime required to execute the temperature control plan on a thermal cycler, (202)
• Determining this at least one rate of temperature change by means of an evaluation program using the temperature plan data and the runtime data; (203)
• Provision of 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)

• Bereitstellen von Temperierplandaten, die die Haltezeit und die Temperatur mindestens einer Temperaturstufe des Temperierplans bestimmen; (201)
• Bereitstellen von Laufzeitdaten, welche die Laufzeit bestimmen, die zur Ausführung des Temperierplans auf einem Thermocycler benötigt wird, (202)
• Ermittlung dieser mindestens einen Temperaturänderungsgeschwindigkeit durch ein die Temperieplandaten und die Laufzeitdaten verwendendes Auswertungsprogramm; (203)
• Bereitstellen der mindestens einen, zuvor ermittelten, Temperaturänderungsgeschwindigkeit für die Steuerung der Temperiervorrichtung des Thermocyclers in Abhängigkeit von dieser mindestens einen Temperaturänderungsgeschwindigkeit. (204)
• Verwenden der mindestens einen, zuvor ermittelten, mindestens einen Temperaturänderungsgeschwindigkeit zur Bestimmung von Steuerungsparametern, welche diese mindestens eine Temperaturänderungsgeschwindigkeit beinhalten und welche einen dem Temperierplan entsprechenden Temperiersteuerungsplan bestimmen; (301)
• Steuern der Temperiervorrichtung mittels der Steuerungsparameter und der elektronischen Steuerungseinrichtung, um diesen Temperiersteuerungsplan unter Verwendung dieser mindestens einen Temperaturänderungsgeschwindigkeit auszuführen. (302)

Figure 5 shows the exemplary method 300 according to the invention for controlling the temperature control device of a thermal cycler, wherein the thermal cycler has the temperature control device for temperature control of a thermoblock receiving 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 one Has control device which is set up to control the temperature control device by means of control parameters. The method 300 here comprises the following steps:

• Providing temperature control plan data which determine the holding time and the temperature of at least one temperature level of the temperature control plan; (201)
• Provision of runtime data which determine the runtime required to execute the temperature control plan on a thermal cycler, (202)
• Determining this at least one rate of temperature change by means of an evaluation program using the temperature plan data and the runtime data; (203)
• Providing the at least one, previously determined, rate of temperature change for controlling the temperature control device of the thermal cycler as a function of this at least one temperature change rate. (204)
• Using the at least one previously determined at least one temperature change rate for determining control parameters which contain these at least one temperature change rate and which 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 (14)

Method (200) for determining at least one rate of temperature change for the control of the temperature control device of a thermal cycler, wherein the control temperature-controls a sample-receiving thermoblock of the thermal cycler for carrying out polymerase chain reactions in these samples according to a temperature control schedule, during which a change is made between temperature levels by changing the temperature with a Temperature change rate is changed, comprising the steps: - Providing temperature control plan data which determine the holding time and the temperature of at least one temperature level of the temperature control plan; (201) - Provision of runtime data that determine the runtime that is required to execute the temperature control plan on a thermal cycler, (202) - Determining this at least one rate of temperature change by means of an evaluation program using the temperature 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) The method of claim 1, wherein the tempering plan data determine at least a first holding time and a first temperature of a first temperature level and at least a second holding time and a second temperature of a second temperature level of the temperature control plan, the first temperature being higher than the second temperature, according to the temperature control plan is switched between temperature levels by starting from the first temperature with a first temperature change rate (cooling rate) and starting from the second temperature with a second temperature change rate (heating rate).
and wherein the evaluation program determines from the temperature control plan data and the runtime data the at least one first temperature change rate that is used as the cooling rate for setting the second temperature level and at least one second temperature change rate that is used as the heating rate for setting the first temperature level,
wherein these at least one cooling speed and at least one heating speed are provided for the control of the temperature control device of the thermal cycler. The method of claim 1 or 2, wherein during a cycle of the tempering schedule there is at least one time interval during which at least one constant temperature change rate is applied and that contains a settling time period which is determined by the period between the presence of the constant temperature change rate and the presence of a temperature step to be set , during which 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, comprising the step: Provision of settling data which contains information about at least one settling time period, the transient data also being used in the determination of this at least one temperature change rate by the evaluation program. The method of claim 3 including the step of: - Providing this transient data by inputting a user on a user interface device of a data processing device by means of which the evaluation program is executed. Method according to one of the preceding claims, wherein the running time includes a latency interval during which, at the beginning of a temperature control plan, a heatable cover, which covers the temperature control block of the thermal cycler containing the samples during the implementation of the polymerase chain reaction, is set to a target temperature, the runtime data also contain the information about this latency interval. Method (300) for controlling the temperature control device of a thermal cycler, the method including that method according to one of claims 1 to 5, which determines the at least one rate of temperature change,
wherein the thermal cycler has the temperature control device for temperature control of a thermoblock holding 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 has an electronic control device which is set up to control the temperature control device by means of control parameters,
and the method comprises the steps of the method according to one of claims 1 to 5 and the following steps: - Using the at least one, previously determined, at least one temperature change rate for determining control parameters which contain these at least one temperature change rate and which 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 carry out this temperature control control plan using this at least one temperature change rate. (302) A method for controlling the temperature control device of a first thermal cycler by simulating the temperature control behavior of a second thermal cycler, the method including that method according to one of claims 1 to 5, which determines the at least one temperature change rate that characterizes the temperature control behavior of the second thermal cycler,
wherein the first thermal cycler is operable at a first maximum temperature rate of change that is a cooling rate or a heating rate, and wherein the second thermal cycler is operable at a second maximum temperature rate of change that is a cooling rate or a heating rate, the first maximum temperature rate of change is greater than or equal to is the second maximum rate of temperature change,
wherein the first thermal cycler has the temperature control device for temperature control of a thermoblock receiving 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 has an electronic control device which is set up to control the temperature control device,
and the method comprises the steps of the method according to one of claims 1 to 5 and the following steps: - Using the at least one, previously determined, at least one temperature change rate for determining control parameters which contain these 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 control this temperature control plan using this at least one temperature change rate to execute. The method of claim 7, wherein the at least one rate of temperature change is less than the first maximum rate of temperature change. Thermal cycler (100), in particular for carrying out polymerase chain fractions in laboratory samples, comprising: a tempering device for tempering a thermoblock receiving the samples according to a tempering plan, during which switching between temperature levels is carried out by changing the temperature at the thermoblock with a temperature change rate; an electronic control device which has 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 plan data that determine the temperature control plan and runtime data that determine the runtime of the temperature control plan, and using the at least one, previously determined, in particular according to the method according to one of claims 1 to 5, at least one temperature change rate for determining control parameters which contain 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 execute this temperature control plan using this at least one temperature change rate. Thermal cycler according to claim 9, wherein the electronic control device is set up in particular to carry out the method according to one of claims 1 to 5, wherein the electronic control device is set up to 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: - Acquisition of tempering plan data which determine the holding time and the temperature of at least one temperature level of the tempering plan; - Collection of runtime data which determine the runtime which is required to execute the temperature control plan on a thermal cycler, - Determination of this at least one rate of temperature change 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. 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, the method according to one of claims 1 to 5 being carried out in particular 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 set up to receive this at least one temperature change rate, in particular also the temperature control plan data and / or the runtime data, via the data connection. The thermal cycler according to claim 9, which has a user interface device, wherein the electronic control device is set up to acquire 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. Program code which carries out the following steps according to the method according to one of claims 1 to 8 when it is carried out by means of a data processing device, in particular the data processing device of an electronic control device of a thermal cycler: - Acquisition of temperature control plan data which are entered in particular by a user via a user interface device which is connected to the data processing device and which is in particular part of the thermal cycler and which determine the holding time and the temperature of at least one temperature level of the temperature control plan; Acquisition of runtime data, which are entered in particular by a user via a user interface device which is connected to the data processing device and is in particular part of the thermal cycler and which determine the runtime which is used to execute the temperature control plan on a thermal cycler, - Determination of this at least one temperature change rate by means of the evaluation program using the temperature control plan data and the runtime data, which is executed in particular 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, at least one temperature change rate for determining control parameters which contain these at least one temperature change rate and which determine a temperature control plan; - Controlling the temperature control device by means of the control parameters and the electronic control device in order to carry out this temperature control control plan using this at least one temperature change rate. Use of the method according to 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.

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