JPS59179073A - Bacteria counter - Google Patents

Abstract

PURPOSE:A device capable of measuring an amount of bacteria in a short time, by irradiating a measurement solution of a reaction product of a specific reagent and bacteria in water with exciting light rays to emit fluorescence, calculating the fluorescence in terms of fluorescent output per bacterium. CONSTITUTION:Acridine Orange (derivative) 4 is added to measruement water which is fed from the extremely pure water line 1 through the branched pipe 3, and is reacted with DNA of bacteria in the reaction chamber 3 to give a substance to produce fluorescence by exciting light irradiation. The reaction solution for measurement is introduced to the measuring chamber 5. The solution for measurement in the measurement chamber 5 is irradiated with the exciting light from the light source 7 through the interference filter 8, the condenser 9 and the chopper 10. Flouorescence is produced depending upon an amount of the reaction product, output of the photomultiplier 12 having received the fluorescence is inputted through the lock-in amplifier 13 to the calculator 14, and an output value corresponding to the amount of bacteria is displayed on the indicator 15. Measurement of the amount of bacteria requiring a long time conventionally can be done in an extermely short time.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a bacteria counter, and particularly to a bacteria counter for measuring the amount of bacteria in ultrapure water used in semiconductor manufacturing and the like.

In water quality testing, in addition to testing the types of bacteria contained in the sample water, it is important to test how many bacteria are present per unit amount of water. This inspection of the amount of bacteria is used, for example, when only an extremely small amount of bacteria is allowed in the production of medical products, or in the production of semiconductor (particularly VLSI semiconductor) elements.
1. It can be said that this test is indispensable in fields that require extremely clean water, such as cases where only an amount of bacteria of about 100 or less per cc is allowed.

Since Cr, the method of testing the amount of bacteria per unit amount of water is to collect the produced ultrapure water, double the bacteria contained therein, multiply it, and then pass it through a filter. A common method is to stain the bacteria, then observe and count the stained bacteria using a microscope.

However, when using this type of testing method, it takes about one hour from sampling ultrapure water to counting the amount of bacteria, so even if the water quality is found to be unsuitable by this test, this testing period will be There is a great possibility that the semiconductor elements and the like manufactured therein will be defective, resulting in an increase in production costs.

Therefore, the present invention aims to provide a bacteria counter that can detect the amount of bacteria in measurement water in an extremely short period of time, and that can reduce the production cost of semiconductors and the like by constantly monitoring the amount of bacteria. This is the purpose.

In order to achieve this purpose, the present invention has been developed by introducing a translucent measuring chamber into which the measuring water containing acrinon orange or its derivatives is added, which produces a reaction product with the bacteria it contains. , an excitation light source for irradiating the measurement water in the 7tl11 window chamber with excitation light to generate fluorescence from the reaction product; a converter for receiving the fluorescence into light T; and a converter for receiving the fluorescence;
1) A bacteria counter characterized in that it has an arithmetic unit for outputting the output of the converter as a fluorescence output per unit bacterium.

Hereinafter, the present invention will be explained in more detail with reference to the drawings. FIG. 1 shows an embodiment of the present invention, where l is a line for ultrapure water led to, for example, semiconductor manufacturing equipment, and 3
Acridine orange or a derivative thereof (for example, 3-amino-6-medoxyacridine, etc.) is supplied from the stain chamber 4 to the measurement water supplied from the ultrapure water line 1 through the branch channel 2. There is a reaction chamber in which acrinon orange or its derivatives are added and reacted. Acridine orange or its derivatives are the DNA of bacteria in the measurement water.
It reacts with A (dioxybo niclear acid, which is the core of the protein) to produce a reaction product, and this product becomes a substance that generates fluorescence when irradiated with excitation light.

For example, as excitation light, approximately 440 to 500 nm (BQ16 approximately 485 nm) is used, and approximately 500 nm to approximately 500 nm
Fluorescence of 80 nm (peak value approximately 530 nm) is generated. 5 is a valve for controlling the supply of ultrapure water and acridine orange or its derivatives to the reaction chamber.

The measurement water in which reaction products have been produced (produced in about 1 to 2 minutes) is then led to the measurement tube 5 and then discharged. The measurement chamber 5 is for measuring the amount of fluorescence produced by irradiating excitation light onto the measurement water produced during the reaction described above to emit fluorescence. It must be at least partially transparent to light in the wavelength range of manual excitation light and fluorescent light, and for example, a quartz tube is used.

7 is an excitation light source, for example, a mercury run fN is used;
Excitation light is irradiated to the measurement water in the measurement chamber 5 through the interference filter 8, the condensing lens 9, and the chopper 10 for irradiating only the excitation light from the fluorescence lifetime of the reaction product. 12 is a photomultiglayer for receiving the fluorescent light generated by irradiating the reaction product in the measurement water with excitation light and generating an output based on the intensity of this fluorescent light. This is an interference filter placed on the incident side of the photomultiplier 12. In FIG. In order to prevent light from entering, the direction perpendicular to the optical axis of the excitation light (
It would be better if they were placed in the straight direction (1) on the paper.

In this way, the output of the photomartin layer 12 which receives the fluorescent light corresponding to the reaction product of the reaction product 5 is inputted to the arithmetic unit 14 via the lock-in amplifier 13. This calculator 14 is used to determine the 'id' of bacteria in the measurement water.
This is to obtain information about bacteria by interpolating the output of the photomultiglare 12 corresponding to The amplitude value of the output from <
The amount of bacteria contained in the constant oil water can be calculated by converting it into the amplitude value per cutiaria.

Here, we will refer to the output value for the 115th position that can be preset in the operator 14. The aforementioned acrinon orange or its derivatives are substances that react uniformly to almost all bacteria contained in water. Furthermore, the reaction product is produced in a specific wavelength range (for example, 530 nm for acridine orange), regardless of the type of bacteria.
It has the property of emitting fluorescent light whose peak wavelength is at rn. Furthermore, although there are over 20 types of bacteria, it is extremely rare for two or more types of bacteria to survive under certain environmental conditions. Therefore, it is easy to set the fluorescence intensity per unit of bacteria in the sample water in advance, and furthermore, it is possible to perform highly accurate calculations just by setting this value once.

15 is a display device for displaying an output value corresponding to bacteria bj obtained by the calculator 14;

16 is a power supply, and 17 is a controller for operating the chopper 101, the photomultislier 12, and the lock-in amplifier 13 in synchronization.

In the above embodiment, for example, 1 ec per 1 ec of the measuring water
In order to accurately detect the amount of bacteria in ultrapure water that contains less than 0.00 bacteria, a chopper that is driven synchronously is used to eliminate the influence of stray light from the outside and detect only fluorescent light. Although lO and a lock-in amplifier 13 are provided, these are not necessarily necessary, and for example, the light amount 7, the measurement chamber 5, and the photomultiplier 12 may be housed in a dark box. Furthermore, if the stray light output such as 31% is a constant flame, the arithmetic unit 14 may remove this stray light output. In cases where the chopper 1o and the lock-in amplifier 13 are not used, the computing unit 14 may be set to convert the integrated output from the photo multiplier 12 into integrated time and output per unit bacteria. .

FIG. 2 shows another embodiment of the present invention, in which a mechanism for detecting the zero level is provided in order to further improve the accuracy of bacterial path detection. In Figure 2, the first
Parts that are the same as those in the drawings are marked with the same sign and their explanation will be omitted. In FIG. 2, a similar measurement chamber 18, photomultiplier 19, and interference filter 20 are provided in parallel with the measurement chamber 5, photomultiplier 12, and interference filter 11 shown in FIG. Measurement water to which acridine orange or its derivatives are not added is introduced into the measurement chamber 18, and the amount of light corresponding to the zero level is detected. 21 and 22 are half mirrors and mirrors for irradiating excitation light from the excitation light source 7 to the measurement chambers 5 and 18 for measurement and zero level detection, respectively. 23 is 2
It is a differential amplifier that receives outputs from two photomultipliers 12 and 19 as inputs. The output from which the zero level has been removed by the differential amplifier 23 is displayed on the display 15 via the lock-in amplifier 13 and the arithmetic unit 14, as in the previous embodiment.

Next, based on the present invention, the number of bacteria in the measurement water is determined by the absorbance when 490 nm peak excitation light is applied to the reaction product obtained by adding a certain amount of acridine orange to the measurement water. Graphs showing an example of the fluorescence intensity and fluorescence intensity are shown in FIGS. 3 and 4. As shown in Figures 3 and 4, the fluorescence intensity varies depending on the amount of bacteria in the measurement water, and the amount of bacteria in the measurement water can be detected by the output based on this fluorescence intensity! '11ro.

As explained and explained in detail above, according to the present invention, the conventional length 'J
Bacteria in the measurement water, which was previously calculated by multiplying I'lj by counting the amount of bacteria, can be carried out in a very short period of time, which can improve the yield of products in semiconductor manufacturing, for example, and reduce raw Pr costs. can contribute to the development of

[Brief explanation of drawings]

1 is a graph showing fluorescent properties used in bright light. In the drawing, 1 is the ultrapure water line, 2 is a branch pipe, 3 is a reaction chamber; 4 is the dye room, 5.18 is the measurement room, 7 is an excitation light source, 8.11.20 is an interference filter, 10 is Jotsupa, 12.19 is the multiplier, 13 is a lock-in amplifier, 14 is a computing unit. Patent u-1 applicant Fuji Photo Film Co., Ltd. agent Patent Attorney Hikari Ishibe (and others) Figure 1 Figure 2 Intersect at 1ogN (high 1)r Figure 4 10, of Day ·liver Sword 2 1oqN (bacteria)

Claims (1)

[Claims] A translucent measurement chamber into which the measurement water containing acridine orange or its derivatives was added and produced a reaction product with the bacteria contained therein, and the measurement water in this measurement chamber was irradiated with excitation light to cause the reaction. An excitation light source for generating fluorescent light from the product, a photoelectric converter for receiving the fluorescent light, and an arithmetic unit for converting the output of the photoelectric converter into a fluorescent output per unit bacterium and outputting the result. A bacteria counter comprising: