FI81211B - Method and device for rapid determination of the form of cells or the like in a flow - Google Patents

Abstract

In current work on the development of mutations of cells and microbes these are subjected to much radiation and chemicals in order to cause the mutation. After such treatment the required cells are selected for individual study usually using microscopes and semi-automatic devices of some kind. In this invention a method has been developed whereby microphotography can be replaced by representing statistically in a rapid way the profile of the cell's or corresponding body's shadow from different directions and constructing a three-dimensional image by computer on the basis of this. This image technique can also be implemented directly with a microcircuit technique and several parallel image-taking channels can be produced for the same sensor.

Description

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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., so that they have 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), one person's working time would be between ten and ten years. Partial automation now achieves speeds of about 5 cells per minute. In this case, humans are still involved in semi-automated cell selection. A technology is being developed that changes 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, from a tube or duct through which they are generally passed by water or some other medium, characterized in that on one side of the duct there are several successive light transmitters (11). ), (12), (13), etc., and on the opposite side in the transverse direction of the channel, linear light intensity meters that measure the projection from different directions statistically by the shadow of the passing body, and these projections from different directions are used to form the size and shape of the body when the flow rate at the orifice is known. A method according to claim 1, characterized in that the channel between the measuring points always widens somewhat and is made so that it is a constant depth engraving in the depth direction of the image and the lid on the engraving can be opened if the channel is blocked for washing. Method according to Claim 1, characterized in that regions outside the visible light (UV and IR regions) are used, the aim being to select a wavelength whose absorption difference between the cell and the surrounding medium is as large as possible. Method according to Claims 1 to 3, characterized in that the flow rate is measured from the channel computationally from the delay of the various measuring points statistically and is calibrated using known bodies. 4,81211 5. Apparatus for rapidly determining the shape of cells or the like, such as microbes, from a tube or channel (1) through which cells flow in a medium, e.g. water, characterized in that light transmitters (11-) are provided on one side of the channel (1). 13), which become mainly parallel light, a plurality of successive (11), (12), (13), etc., and on the opposite side in the transverse direction of the channel, linear light intensity meters (21-23) which measure the projection from different directions statistically by the shadow of the passing body. , and these projections from different directions are used by the computer to form the size and shape of the body when the flow rate at the orifice is known. Device according to Claim 5, characterized in that the channel (1) widens between the measuring points. Device according to Claim 5 or 6, characterized in that the channel (1) is provided with an openable lid for easy cleaning. 6 81211