Method for controlling a thermal cycler, and thermal cycler

Abstract

The invention relates to a method for controlling a thermal cycler and a thermal cycler, in which the determination of at least one temperature change rate is carried out by an evaluation program using tempering schedule data and run time data, whereby in particular the tempering behavior of a slower thermal cycler can be simulated on a faster thermal cycler.

Claims

1. Method (200) for determining at least one temperature change rate for controlling the tempering device of a thermal cycler, in which the control tempers a samples-receiving thermal block of the thermal cycler for performing polymerase chain reactions in those samples according to a tempering schedule, during which the temperature is changed between temperature levels by changing the temperature with a temperature change rate, comprising the steps: Providing tempering schedule data that is determining the hold time and the temperature of at least one temperature step of the tempering schedule; (201) Providing run time data that is determining the run time required for the execution of the tempering schedule on a thermal cycler, (202) Determination of the at least one temperature change rate by means of an evaluation program using the tempering schedule data and the run time data; (203) Providing the at least one, previously determined, temperature change rate for controlling the tempering device of the thermal cycler in function of said at least one temperature change rate. (204)

2. Method according to claim 1, in which the tempering schedule data determines at least a first hold time and a first temperature of a first temperature step and at least a second hold time and a second temperature of a second temperature step of the tempering schedule, in which the first temperature is higher than the second temperature, in which the temperature is changed between temperature levels according to the tempering schedule by cooling at a first temperature change rate (cooling rate) starting from the first temperature and by heating at a second temperature change rate (heating rate) starting from the second temperature. and in which the at least one first temperature change rate, which is used as cooling rate for the adjustment of the second temperature level, and the at least one second temperature change rate, which is used as heating rate for the adjustment of the first temperature level, are determined by the evaluation program from the tempering schedule data and the run time data, in which these at least one cooling rate and at least one heating rate are provided for controlling the tempering device of the thermal cycler.

3. Method according to claim 1 or 2, in which there is at least one time interval in a cycle of the tempering schedule, during which at least one constant temperature change rate is applied and which comprises a period of the transient oscillation that can be identified as the time between the presence of the constant temperature change rate and the presence of a temperature level to be adjusted, in which the control of the tempering device of the thermal cycler performs a transient oscillation during this time interval, which is part of the temperature control of a thermal cycler, comprising the step: Providing transient oscillation data that comprises information on at least one period of the transient oscillation, in which also the transient oscillation data is used in the determination of that at least one temperature change rate by the evaluation program.

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

5. Method according to any one of the preceding claims, in which the run time includes a latency interval, during which, at the beginning of a tempering schedule, at first a heatable lid covering the tempering block of the thermal cycler that contains the samples during the performance of the polymerase chain reaction is adjusted to a set temperature, in which the run time data also includes the information on this latency interval.

6. Method (300) for controlling the tempering device of a thermal cycler, in which the method comprises that method according to any one of claims 1 to 5, which determines the at least one temperature change rate, in which the thermal cycler comprises the tempering device for tempering a sample-receiving thermal block for performing polymerase chain reactions in these samples according to the tempering schedule defined by the method according to any one of the claims 1 to 5, and comprises an electronic control device that is configured for controlling the tempering 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, temperature change rate for the determination of control parameters, which comprise said at least one temperature change rate, and which determine a tempering control schedule corresponding to the tempering schedule; (301) Controlling the tempering device by means of the control parameter and the electronic control device in order to execute the tempering control schedule using said at least one temperature change rate. (302)

7. Method for controlling the tempering device of a first thermal cycler by simulating the tempering behavior of a second thermal cycler, in which the method includes that method according to any one of claims 1 to 5, which determines at least one temperature change rate that characterizes the tempering behavior of the second thermal cycler, in which the first thermal cycler can be operated with a first maximum temperature change rate, which is a cooling rate or a heating rate, and in which the second thermal cycler can be operated with a second maximum temperature change rate, which is a cooling rate or a heating rate, in which the first maximum temperature change rate is greater than or equal to the second maximum temperature change rate, in which the first thermal cycler comprises the tempering device for tempering a sample-receiving thermal block for performing polymerase chain reactions in these samples according to the tempering schedule defined by the method according to any one of claims 1 to 5, and comprises an electronic control device that is configured for controlling the tempering 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, temperature change rate for the determination of control parameters, which comprise said at least one temperature change rate, and which determine a tempering control schedule corresponding to the tempering schedule; Controlling the tempering device by means of the control parameters and the electronic control device in order to execute the tempering control schedule using said at least one temperature change rate.

8. Method according to claim 7, in which the at least one temperature change rate is smaller than the first maximum temperature change rate.

9. Thermal cycler (100), in particular for performing polymerase chain reaction in laboratory samples, comprising: a tempering device for tempering a sample-receiving thermal block according to a tempering schedule, during which the temperature is changed between temperature levels by changing the temperature at the thermal block with a temperature change rate; an electronic control device that comprises a data processing device and that is configured for controlling the tempering device in order to execute the following steps: Using tempering schedule data determining the tempering schedule, and run time data determining the run time of the tempering 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, for the determination of control parameters, which comprise said at least one temperature change rate, and which determine a tempering control schedule corresponding to the tempering schedule; Controlling the tempering device by means of the control parameter and the electronic control device in order to execute the tempering control schedule using said at least one temperature change rate.

10. Thermal cycler according to claim 9, in which the electronic control device is configured in particular for the execution of the method according to any one of claims 1 to 5, in which the electronic control device is configured to execute an evaluation program using the data processing device of the control device, and in which the electronic control device is configured to execute the following steps: Acquiring of tempering schedule data that is determining the hold time and the temperature of at least one temperature step of the tempering schedule; Acquiring of run time data that is determining the run time required for the execution of the tempering schedule on a thermal cycler, Determination of the at least one temperature change rate by means of the evaluation program using the tempering schedule data and the run time data; Using the at least one temperature change rate determined by the evaluation program for controlling the tempering device of the thermal cycler in function of said at least one temperature change rate.

11. Thermal cycler according to claim 9, in which the data processing device of the electronic control device comprises an interface device, by means of which a data connection with an external data processing device can be established, in which the method according to any one of claims 1 to 5 is executed in particular on this external data processing device, in order to provide at least one temperature change rate, in which the data processing device of the electronic control device is configured to receive this at least one temperature change rate, in particular also the tempering schedule data and/or the run time data, via the data connection.

12. Thermal cycler according to claim 9 comprising a user interface device, in which the electronic control device is configured to acquire the tempering schedule data entered by a user via the user interface device, and to acquire the run time data entered by a user via the user interface device.

13. Program code that executes the following steps according to any one of claims 1 to 8 if 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: Acquiring tempering schedule data that is in particular entered by the user via a user interface device connected to a data processing device, which in particular part of the thermal cycler, and that is determining the hold time and the temperature of at least one temperature step of the tempering schedule; Acquiring run time data that is in particular entered by the user via a user interface device connected to a data processing device, which in particular is part of the thermal cycler, and that is determining the run time required for the execution of the tempering schedule on a thermal cycler, Determination of said at least one temperature change rate by means of the evaluation program using the tempering schedule data and the run time data, and being executed in particular by the data processing device; Providing the at least one temperature change rate determined by the evaluation program for controlling the tempering device of the thermal cycler in function of said at least one temperature change rate; Using the at least one, previously determined, temperature change rate for the determination of control parameters, which comprise said at least one temperature change rate, and which determine the tempering control schedule; Controlling the tempering device by means of the control parameter and the electronic control device in order to execute the tempering control schedule using said at least one temperature change rate.

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

Description

[0082] Preferred embodiments of the thermal cycler according to the present invention can be inferred in particular from the description of one of the methods according to the invention. Preferred embodiments of the method according to the present invention can be inferred in particular from the description of the thermal cycler according to the present invention. Other preferred embodiments of the method and the thermal cycler according to the present invention can be inferred from the description of the embodiment examples according to the figures.

[0083] In the figures:

[0084] FIG. 1a depicts a perspective front view of a thermal cycler according to the present invention in an embodiment example.

[0085] FIG. 1b depicts a perspective back view of the thermal cycler from FIG. 1a.

[0086] FIG. 2a to 2e each depict screen contents, which can be displayed on the screen of the thermal cycler from FIGS. 1a and 1b.

[0087] FIG. 2e depicts a screen input dialog, in which the user can enter the known run time of the tempering schedule after having entered the tempering schedule. The thermal cycler autonomously calculates the temperature change rates from the run time.

[0088] FIG. 3a depicts an example of a tempering schedule defined by the user, which is defined in the thermal cycler or the method according to the present invention in particular by the tempering schedule data.

[0089] FIG. 3b schematically depicts a tempering control schedule that is calculated by the thermal cycler or the method according to the present invention from the run time and the tempering schedule data from FIG. 3a.

[0090] FIG. 3c schematically depicts the temperature profile when changing between two temperature levels, indicating the effective cooling rate as difference quotient and the maximum cooling rate as maximum differential.

[0091] FIG. 4 schematically depicts the process sequence of an example of the method according to the present invention for determining at least one temperature change rate from the tempering schedule data and the run time data.

[0092] FIG. 5 schematically depicts the process sequence of an example of the method according to the present invention for controlling a thermal cycler using the steps of the method for determining at least one temperature change rate from the tempering schedule data and the run time data from FIG. 3.

[0093] FIG. 1a depicts a perspective front view of a thermal cycler 100 according to the present invention in an embodiment example. On the outside, the thermal cycler 100 is characterized by a lid handle 1 for closing and opening the heating lid, the heating lid 2, the heating plate 3 that is located in the heating lid and that can be heated to ca. 105° C. for avoiding condensation on the insides of the sample containers, the aluminum thermal block 4 with (here) 385 intakes for taking in PCR containers, in particular a 384 microwell plate, which is contacted from underneath (not visible) with six Peltier elements that constitute the tempering elements of the tempering device of the thermal cycler for heating and cooling the thermal black and that are contacted at their undersides (not visible) by a heat sink to dissipate the excess heat of the heat pumps to the surroundings, a mains connection socket with mains switch 5, a connection socket for Ethernet 6, a connection socket for the data exchange with another thermal cycler 7, a cover 8 for covering a USB socket, a touchscreen 9 operating as user interface device, a name plate 10. Alternatively, a 96 Aluminum or Silver block can be used as exchangeable thermal block.

[0094] The thermal cycler 100 comprises a control device with a program-controlled microprocessor (not depicted), that is configured for executing the steps of the methods 200 and 300 according to the present invention in that the control program of the thermal cycler 100 is programmed to be able to execute these steps.

[0095] FIG. 3a depicts a typical tempering schedule that could have been defined by the user via the touch screen 9 (for example, see FIG. 2a). It comprises the desired (here: three) temperature steps 95° C., 65° C., 72° C. and their hold times Δt1, Δt2, Δt3 of one cycle that is to be repeated successively 30 times (“×30”).

[0096] FIG. 3b depicts a typical tempering control schedule, which, here, has been calculated by the control device of the thermal cycler from the tempering schedule data according to FIG. 3a and the run time data entered by the user. The tempering control schedule comprises the time intervals Δt.sub.s_cool, Δt.sub.s_heat1 and Δt.sub.s_heat2. In the time interval Δt.sub.s_cool the average temperature measured at the tempering block by the temperature sensors of the tempering device is cooled from 95° C. to 65° C., for example with a cooling rate of 1.0° C./sec, which corresponds to the maximum cooling rate of an older, previously used thermal cycler TC.sub.x that, for executing, required the run time now entered by the user. Correspondingly, in the time interval Δt.sub.s_heat1 the temperature is raised from 65° C. to 72° C., e.g at a heating rate of 2.0° C./sec, which corresponds to the maximum heating rate of the older, previously used thermal cycler TC.sub.x that, for executing, required the run time now entered by the user. In the time interval Δt.sub.s_heat2 the temperature is raised from 72° C. to 95° C., e.g. at a heating rate of 2,0° C./sec, so that the cycle can start over. The thermal cycler according to the present invention exhibits a greater maximum heating and cooling rate, namely 10° C./sec and 5° C./sec, so that it can easily execute the calculated heating and heating and cooling rates of the older devices. The duration of the transient oscillation according to a standard transient control is also included in the time interval in each case. As a result, the thermal cycler according to the present invention simulates the tempering behavior of the older apparatus, so that the user can reproduce the previously performed reaction protocols without any problems. In this way, migration from an older apparatus to a thermal cycler according to the present invention is facilitated.

[0097] FIG. 4 shows an example of the method 200 according to the present invention for determining at least one temperature change rate for controlling the tempering device of a thermal cycler by calculating at least one temperature change rate from the know runtime of a known tempering schedule. The method 200 comprises the following steps: [0098] Providing tempering schedule data that is determining the hold time and the temperature of at least one temperature step of the tempering schedule; (201) [0099] Providing run time data that is determining the run time required for the execution of the tempering schedule on a thermal cycler, (202) [0100] Determination of the at least one temperature change rate by means of an evaluation program using the tempering schedule data and the run time data; (203) [0101] Providing the at least one, previously determined, temperature change rate for controlling the tempering device of the thermal cycler in function of said at least one temperature change rate. (204)

[0102] FIG. 5 shows an example of the method 300 according to the present invention for controlling a tempering device of a thermal cycler, in which the thermal cycler comprises the tempering device for tempering a sample-receiving thermal block in order to perform polymerase chain reactions in these samples according to the tempering schedule defined by the method according to any one of claims 1 to 5, and comprises an electronic control device that is configured for controlling the tempering device by means of control parameters. Here, the method 300 comprises the following steps: [0103] Providing tempering schedule data that is determining the hold time and the temperature of at least one temperature step of the tempering schedule; (201) [0104] Providing run time data that is determining the run time required for the execution of the tempering schedule on a thermal cycler, (202) [0105] Determination of the at least one temperature change rate by means of an evaluation program using the tempering schedule data and the run time data; (203) [0106] Providing the at least one, previously determined, temperature change rate for controlling the tempering device of the thermal cycler in function of said at least one temperature change rate. (204) [0107] Using the at least one, previously determined, temperature change rate for the determination of control parameters, which comprise said at least one temperature change rate, and which determine a tempering control schedule corresponding to the tempering schedule; (301) [0108] Controlling the tempering device by means of the control parameter and the electronic control device in order to execute the tempering control schedule using said at least one temperature change rate. (302)