FI81211C - Method and apparatus for quickly determining the shape of cells or correspondingly in a stream - Google Patents

Description

1 81211

Method and apparatus for the rapid determination of the shape of cells or the like from a flow - Förfarande och anordning för snabb bestämning av formen pä Celler eller motsvarande i en Ström.

One very interesting area of biotechnology is to inherit plants, fungi, bacteria, etc., to give them certain desired properties. For example, a microbe is made to produce a large amount of desired antibiotics. One widely used technique is to expose the cell to radiation or chemicals that cause mutations. The methods yield, with probabilities of 10 "^ - 10"? The desired mutants, which can then be produced in large quantities and further processed.

In the case of microbes, the aim is to separate the individual stem cells by diluting the solution containing them until there is a high probability that there is only one microbe in a given volume. In this case, in practice, a situation is reached in which about half of the standard volumes taken at the start of cultivation do not contain a microbes at all and the yield efficiency of the cultures decreases correspondingly. The dilution procedure is not suitable for plant cells, but healthy desired individual cells must be selected from the cell mass using a microscope for separate growth. If these cells are selected from the microscope plate with a micropipette for separate growth at a specific location on the medium, one may reach a rate of 1 cell per minute. According to such a typical probability (10 "5 - 10-6), the working time of one person would be from ten to ten years. Based on its visual observations, the computer selects and diverts the flow with a tap to the desired tube, from where the desired cell is automatically collected for single growth. wavelength used, because it is made to the eye at a wavelength that is only part of the electromagnetic radiation that complies with the laws of optics. the monitoring and evaluation criteria in the various fermentation processes of biotechnology are the monitoring of the nature and quality of the cells and other particles contained in the production solution. Thus, e.g. the viability of the organisms used in the process and the presence of various unwanted particles. This is done for small objects from microscopically taken samples at certain intervals. The hardware described here would allow for continuous monitoring, making control crucial.

The object of the invention is achieved by a method and a device, the features of which are set out in the claims.

The main advantages of the present invention are the low cost of the optics optics compared to the microscope-camera combination, and several of these can be placed in parallel very advantageously, since the light sources and the data processing can be combined. Also, the wavelength used is not limited to the visible range, but suitable wavelengths from the IR and UV ranges can also be used, making sorting easier. Also in UV areas, sharper optical boundaries are obtained because the wavelength decreases.

In the following, the invention is presented in more detail by means of the principle diagram 1. The image is greatly enlarged because the diameters of the cells or microbes are generally in the range of 0.1 to 0.001 mm and the length / width ratio of the tube is increased so that the length is proportionally increased less than the width.

The fluid containing the cells enters the container by pressure from the right side of the image into a tube or chamber (2), which is constant in thickness in the depth direction of the image and can always expand in the width direction between the 3 31211 imaging sites to enhance turbulence and reduce flow resistance. Light is introduced into the tube by optical cables (11), (12), (13), etc., the light is measured on the opposite side in a very linear manner, i.e. about 1/10 of the minimum diameter of the cells to be measured, for example by photodiode (21), (22), (23), etc. enters the first shooting location (4) and bypasses it, a signal according to Fig. (31) is obtained from the light intensity. Correspondingly, as the cell passes the next imaging point (5), (6), etc., shadow signals (32), (33), etc. are obtained from different projections of the cells as the cell rotates statistically. When signals are taken at about 10 shooting locations and the signals are digitized, a cell outline can be drawn on a computer, and its shape can be identified. The flow rate can be determined from the outgoing-outflow in many different ways, e.g. by calculation when measuring the flow mass in a unit of time. Once the desired cell or microbe is identified, it is directed to the individual culture after the analysis channel by means of a fast-acting tap or the like.

The method is not limited to the cellular and microbial applications disclosed, but may vary within the scope of the claims.

Claims (7)

A method for rapidly determining the shape of cells or the like, such as microbes in a tube or channel, through which they are set to roam kindly by water or some other medium, characterized in that on one side of the channel there are light transmitters from which approximately parallel light, several in succession (11), (12), (13), etc. and on the opposite side in the transverse direction of the channel intensity meter for line-shaped light which measures a projection from statistically different directions by means of the shadow of the passing body, and with the aid of these projections obtained from different directions, the size and shape of a computer are formed, where the flow velocity at the opening is known. 2. A method according to claim 1, characterized in that the channel between the measuring points is always extended to some men and made such that it is in the direction of the depth of the image an engraving, and a lid on top of the engraving can be opened for washing, if the channel becomes clogged. . 3. A method according to claim 1, characterized in that the outer regions of the visible light (the UV and IF regions) are used, with the aim of choosing a path length, where the absorption difference between the cell and the surrounding medium is possible. Method according to claims 1-3, characterized in that the flow rate is measured statistically from the delay of the arithmetically different measuring points in the channel and calibrated using known bodies. Device for rapidly determining the shape of cells or the like, such as microbes in a tube or channel (1), through which the cells flow in a medium such as water, characterized in that on one side of the channel (1) there is light - transmitters (11-13), from which coirans essentially parallel light, several in succession (), (12), (13), etc. and on the opposite side in the transverse direction of the channel intensity meter for line-shaped light (21-23), which measures a projection from statistically different directions by means of the shadow of the passing body, and with the aid of these projections obtained from different directions, is formed with a computer body size and shape, where the flow rate at the orifice is known. Device according to claim 5, characterized in that the channel (1) extends between the measuring points. Device according to Claim 5 or 6, characterized in that the channel (1) is provided with a lid which can be opened to facilitate cleaning.