Field of the invention

The invention relates to a method for biological analysis, utilizing equip¬ ment comprising at least a culture unit with controllable conditions, such as a perfusion unit, an incubation unit, or the like, with a substrate arrangement for culturing cells, cell samples and/or a population.

Background of the invention

At present, the monitoring of cells and possible other measures on them are always performed manually. Firstly, this has the result that a so-called individual monitoring and analysis of cells is not carried out in a larger population, unless a cell marker is used. However, the use of a cell marker involves additional steps, thereby increasing the quantity of manual work needed. On the other hand, homogeneous cell popula¬ tions are normally used, because cell markers affect not only the desired target cell but all cells in the cell population. Consequently, when heterogeneous cell populations are used, it is difficult to deter¬ mine what type of cells are marked with the cell marker. Furthermore, the cell markers used may be detrimental to the cells as such, or may cause distorted measurement results in another way.

Description of the invention

It is an aim of the method according to the present invention to provide a decisive improvement to the above-presented problems and thereby to substantially improve the state of art in the field.

To attain this purpose, the method according to the invention is primar¬ ily characterized in that in the method, the parameters to be monitored for the analysis are determined simultaneously for each single cell, cell sample and/or population, including for example the number of cells, the type of cells, the cell division, the time involved in the cell cycle, the death of cells (necrosis, apoptosis), the movement of cells, other mor- phological properties of cells, the cell pedigree and/or the like; and several parameters to be specified are analyzed simultaneously on the basis of the imaging data.

The most significant advantages of the method according to the inven¬ tion include the simplicity and efficiency of its principle and use, wherein a compact analyzing apparatus functioning according to the method makes it possible to work in a way which is considerably more efficient than conventional analyzing techniques. Thanks to the auto- mation and mechanization of the method, it is possible to reduce the work further to a fraction of that performed today. Furthermore, the number of materials used and chemicals required will be reduced. On the other hand, the method according to the invention also makes it possible to monitor heterogeneous populations by analyzing cells/cell samples and or a cell population/cell populations in substrate arrange¬ ments, such as culture sites on a well plate, by a simultaneous analysis of, for example, six predetermined parameters and/or processes to be monitored. This can be implemented by using an imaging arrangement in the culture unit, by recording the cells/cell samples and/or a cell population/cell populations in the substrate arrangements mechanically and systematically at regular intervals to detect morphological, dynamic, kinematic and/or corresponding changes and/or processes therein. The method according to the invention thus makes a fully automatic instrumentation possible, wherein all the functions can be controlled centrally by means of software in a data processing unit. One essential advantage of the method according to the invention also lies in avoiding extra work involved in conventional manual arrange¬ ments, particularly in marking. Another essential advantage is the fact that the parameters measured at different time points are measured from the same cells/cell samples and/or population/populations, which reduces fluctuations in the measuring results.

Advantageous embodiments of the method according to the invention will be presented in the respective dependent claims. The invention also relates to an apparatus functioning according to the method, as defined in more detail in the preamble of the independent claim relating to the apparatus. The primary characteristics of the appa¬ ratus will be presented in the characterizing part of the respective claim.

The most significant advantages of the apparatus according to the invention include the simplicity and reliability of its assembly and use, as well as the possibility to make the process of biological analysis considerably more efficient for a variety of uses. The efficiency of the apparatus according to the invention is based, first of all, on the imag¬ ing arrangement by which the morphological, dynamic, kinematic and/or corresponding changes in the single cells/cell samples making up the heterogeneous population can be analyzed by mechanical imaging at regular intervals. By means of the imaging, it is also possi¬ ble to monitor the development of the cells/cell samples as a function of time for the several parameters to be monitored by analyzing the successive imaging data recorded of the cells/cell samples by means of software in the data processing unit of the apparatus. Thus, by means of the apparatus according to the invention, it is possible to automatically perform all the functions relating to culturing, imaging and analysis in such a way that the risks and disadvantages related to con¬ ventional manual operations can be avoided. In this respect, the appa¬ ratus further comprises, in an advantageous embodiment, an actuator arrangement which automatically changes the culture medium at regular intervals or, if necessary, adds drugs, chemicals, markers or the like in the substrate arrangements, wherein thanks to mechanical operations, extra work is avoided when changing the culture medium.

Advantageous embodiments of the apparatus according to the inven¬ tion will be presented in the respective dependent claims.

The invention relates to a method for biological analysis, utilizing equip¬ ment comprising at least a culture unit with controllable conditions, such as a perfusion unit, an incubation unit, and/or the like, with a sub¬ strate arrangement for culturing cells, cell samples and/or a population. Cells, cell samples and/or a population/populations in a substrate arrangement are automatically imaged by means of an imaging unit in connection with the culture unit, after which they are automatically analyzed by a data processing unit in connection with the culture unit, to monitor predetermined parameters individually according to the imaging data.

The substrate arrangement comprises several culture sites which are imaged at regular intervals mechanically by means of an imaging arrangement belonging to the culture unit, to detect morphological, dynamic, kinematic, and/or corresponding changes and processes in cells/cell samples constituting a homogeneous or heterogeneous population.

In an advantageous embodiment, the cells are analyzed by monitoring the development of single cells/cell samples in the same population by analyzing successive imaging data on them to observe the develop¬ ment as a function of time with respect to the parameters to be moni¬ tored.

Furthermore, in another advantageous embodiment, the cells of a het¬ erogeneous or homogeneous population are analyzed by analyzing the type and phase of a single cell/cell sample, such as the the cell divi¬ sion, the time involved in the cell cycle, the death of cells (necrosis, apoptosis), the movement of cells, or the morphological properties of cells.

Furthermore, in another advantageous embodiment, the cells are ana¬ lyzed without a need to select the cells/cell samples manually before- hand.

Furthermore, in another advantageous embodiment, the cells are ana¬ lyzed without cell markers.

Furthermore, in another advantageous embodiment, the cells are ana¬ lyzed on the basis of the morphology and/or structures of the cells. Furthermore, in another advantageous embodiment, the cell morphol¬ ogy and/or structures are taught to the data processing unit.

Further, in another advantageous embodiment, the analysis is con¬ trolled by software in a data processing unit which is at least in a data transmission connection with the culture unit and in which the parameters to be monitored simultaneously for each single cell/cell sample are determined in advance, including the number of cells, the type of cells, the cell division, the time involved in the cell cycle, the death of cells (necrosis, apoptosis), the movement of cells, other mor¬ phological properties of cells, the cell pedigree and/or the like.

Furthermore, in an advantageous embodiment in practice, the culture medium used in perfusion or incubation is also changed at predeter¬ mined intervals automatically by an actuator arrangement in the appa¬ ratus.

Moreover, drugs, chemicals, cell markers and/or the like are advanta- geously added by the actuator arrangement into the substrate arrangement, such as the culture medium, after which the area of each cell/cell sample to be examined at a time is defined again for the imaging arrangement, if necessary.

Furthermore, in an advantageous embodiment, the substrate arrange¬ ment used is one or more well plates consisting of wells including sev¬ eral culture sites.

The invention also relates to an apparatus applying the method and comprising, first of all, a culture unit with controllable conditions, such as a perfusion unit, an incubation unit, or the like, with a substrate arrangement for culturing cells, cell samples and/or a population. The apparatus comprises, in connection with the culture unit, an imaging unit for imaging the cells, cell samples and/or a population/populations in the substrate arrangement at predetermined intervals by an imaging arrangement therein. In an advantageous embodiment, the apparatus comprises a data processing unit which is at least in a data transmission connection with the culture unit and in which the parameters to be monitored simulta- neously for each single cell, cell sample and/or cell population/cell populations have been determined in advance, including the number of cells, the type of cells, the cell division, the time involved in the cell cycle, the death of cells (necrosis, apoptosis), the movement of cells, other morphological properties of cells, the cell pedigree and/or the like.

Furthermore, in an advantageous embodiment, the apparatus com¬ prises an actuator arrangement for changing the culture medium used for incubation and/or perfusion and/or for adding drugs, chemicals, cell markers and/or the like into the substrate in the substrate arrangement, such as one or more well plates consisting of wells comprising several culture sites, automatically at predetermined intervals.

It is obvious that the invention is not limited to the applications pre- sented or explained above, but it can be modified within the scope of the basic idea of the invention according to each use. It is thus possible to use, firstly instead of well plates as the substrate arrangement, other types of constructions from which the cells/cell samples can be imaged in an unobstructed way. The machine vision of the imaging arrange- ment can also be implemented by various digital imaging/video arrangements by using a variety of techniques for generating the images as such.