JPH0535377B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of the Invention The present invention relates to a sample component quantification device for quantifying samples such as proteins.

(b) Prior art and its problems Conventionally, in quantifying sample components, a sample is applied to a support, the sample components are separated by electrophoresis or chromatography, and then dyed. Then, the optical density signal of the band or spot of the stained sample component is detected with a photomultiplier tube or imaged with an imaging device such as a TV camera, and the image information is analyzed with a computer to determine the sample component. I was trying to quantify it.

However, with this method, separated images of separated sample components range from a small amount to a large amount, and their shapes range from simple to complex. Therefore, when quantifying this separated image,
The problem with the photomultiplier tube mentioned above is that it takes a long time to scan mechanically.
Furthermore, measurement may be difficult depending on the shape.

On the other hand, in the imaging device such as the TV camera mentioned above,
Since the optical density range that can be quantified with a certain sensitivity is narrow, it was necessary to adjust the sensitivity to fall within the proportional response range (dynamic range) in order to quantify a large amount of sample components. Therefore, in this case, a calibration curve must be determined and quantified, and there is a problem that a multi-component sample cannot be easily quantified.

(c) Purpose of the Invention The present invention has been made in view of the above-mentioned problems, and it is possible to improve imaging means such as TV cameras by expanding the optical density range that can be quantified with a constant sensitivity using a transmission means such as an optical filter. The object of the present invention is to provide a sample component quantification device that can simultaneously analyze multiple components of a sample.

(d) Structure and effects of the invention In order to achieve the above-mentioned objects, the present invention has the following features:
A sample, a support on which the sample is coated and separated into each component and dyed with the sample components, an irradiation means for irradiating the support with light, and multiple types with different transmittances depending on the wavelength of the light. a transmitting means, an imaging means for capturing an image of light that is irradiated onto the support and transmitted through the transmitting means, and a calculation means for calculating an image output of the imaging means based on a transmittance of the transmitting means; and display means for displaying the output of the calculation means, and the imaging means switches to a transmission means with an appropriate transmittance according to the optical density of the light from the support, so that the imaging means transmits the transmitted light at a constant sensitivity. The apparatus is configured to take an image of each component of the sample using an imaging means and to quantify each component of the sample.

Therefore, according to the sample component quantification device of the present invention, the transmitting means has an appropriate transmittance depending on the optical density of the light from the support, and the imaging means images an image of the transmitted light with a constant sensitivity. Therefore, sample components larger or smaller than a predetermined optical density range can be converted into a predetermined optical density range by a transmission means such as an optical filter, and imaged by an imaging means, and can be imaged by a TV camera. It is possible to quantify sample components in a wide optical density range within the dynamic range of imaging means such as.

Moreover, since there is no need to adjust the sensitivity of the imaging means depending on the sample component, quantification can be performed quickly and easily.Furthermore, multiple components can be analyzed simultaneously, making it even faster and more versatile. Automatic quantification becomes possible. Therefore, it exhibits remarkable effects in research fields or applied fields such as testing.

(E) Description of Embodiments Hereinafter, embodiments of the present invention will be described in detail based on the drawings.

As shown in FIG. 1, reference numeral 1 denotes a sample component quantification device, which quantifies biological sample components such as proteins.

This sample component quantification device 1 is composed of an optical system 2 and a signal processing system 3, and this optical system 2 includes a support 4.
The light L is irradiated to the area.

Although not shown, this support 4 is coated with a sample, and each component is separated and stained by electrophoresis or chromatography.

Light L from the optical system 2 is irradiated onto the support 4 from a light source 5 (irradiation means) via a filter 6, and is reflected upward. A rotary filter 7 is provided above the support 4. This rotary filter 7 includes a first optical filter 8 on a rotary plate 7a.
a, a second optical filter 8b (transmission means) is attached, and is connected to a pulse motor 9 via a shaft 7b. Each of these optical filters 8a,
The transmittance of the filter 8b differs depending on the wavelength λ of the light L, and both optical filters 8a and 8b are configured so that the wavelength λ of the light L transmitted therethrough is different. The light L transmitted through each of the optical filters 8a and 8b is transmitted through a lens 10 to an imaging device 11 (imaging means) of the signal processing system 3.
It is designed to be incident on .

This imaging device 11 is a single TV camera, and is configured to capture an image of the light L of the support 4 on a light-receiving surface, and to output an image signal. There is. This image signal is transmitted to amplifier 1
2, the analog quantity is converted into a digital quantity by the A/D converter 13, and then input to the switch 14.

The output signal of the control circuit 15 that controls the pulse motor 9 is inputted to this switch 14, and the pulse motor 9 is driven, that is, each optical filter 8a,
8b, the image signals of the imaging device 11 are selected and output to and stored in the plurality of memories M1, M2, . . . Mn. This memory M1, M2,...
The output signal of Mn is input to the calculation device 16 (calculation means) and is calculated. This arithmetic unit 16 is configured to perform correction calculations on the image signal according to the transmittance of each optical filter 8a, 8b, and the corrected image signal is converted from a digital quantity into an analog quantity by a D/A converter 17. After that, it is displayed as a composite screen on the monitor TV 18 (display means).

Further, the output signal of the arithmetic unit 16 is transmitted to the printer 19.
(Display means) is also configured to be input and displayed.

Note that 20 is an input device of the arithmetic unit 16.

Next, the quantitative operation of this sample component quantitative determination device 1 will be explained.

First, the support 4 is coated with a sample, each component is separated by electrophoresis or chromatography, and then dyed, and the support 4 is installed.

Subsequently, light L is emitted from the light source 5, and this light L is irradiated onto the surface of the support body 4 through the filter 6, and the reflected light L is transmitted through the optical filter 8a or 8b and the lens 10 to the imaging device 11. to form an image.
That is, images of each sample component on the support body 4 are captured by the imaging device 11 .

Regarding both optical filters 8a and 8b, the light L
Since the transmittance T differs depending on the wavelength λ, the pulse motor 9 is controlled and driven by the control circuit 15 using the reflected light L to switch.

For example, a protein sample is electrophoresed in Support 4 and Comassie Brilliant Blue R-250
(CBBR-250), this protein sample is specifically stained. And this CBBR−
The maximum absorption wavelength (λmax) of 250 is 552nm and 585nm.
Alternatively, since the wavelength is 590 nm, if it is within the dynamic range of the imaging device 11, the first optical filter 8a having a good transmittance T of the light L around this wavelength (λmax) is used. That is, as shown in FIG. 2, using the first optical filter 8a having a high transmittance T for light L having a wavelength of 500 nm to 600 nm, the imaging device 11 images the optical density signal transmitted through the first optical filter 8a. do.

Furthermore, in the case of light L that is out of the dynamic range of the imaging device 11, as shown in FIG.
Using the second optical filter 8b having a low transmittance T of light L in the range of 500 nm to 600 nm, the imaging device 11 images the optical density signal within the dynamic range transmitted through the second optical filter 8b with the same sensitivity.

In addition, when quantifying, a calibration curve showing the correlation between protein concentration and optical density is used, or a certain protein is used as an internal standard, which has been measured in advance, and quantification is carried out based on these.

The image signal captured by the imaging device 11 is amplified by the amplifier 12, converted to a digital quantity by the A/D converter 13, and then transferred to the control circuit 1 by the switch 14.
5 is selected corresponding to each optical filter 8a, 8b and stored in the memories M1...Mn.

Subsequently, the image signals stored in the memories M1...Mn are corrected and calculated in the arithmetic unit 16 based on the transmittance T of each optical filter 8a, 8b.
It is output to the printer 19 and displayed. Also,
After being converted into an analog quantity in the D/A converter 17, it is displayed on the monitor TV 18 as a composite screen.

This allows the sample components to be quantified.

Figures 4a and 4b show other rotating filters 21 and 22, in which three types of optical filters 8a, 8b, 8c or four types of optical filters 8a, 8b, 8c, 8d are attached to rotating plates 21a and 22a. It is composed of Furthermore, five or more types of optical filters 8 may be attached, and multiple types of ND filters having different amounts of transmitted light may be used.

In addition, in the embodiment, the light L from the light source 5 was reflected by the support 4, but the support 4 was placed on a light box or the like, and the light L transmitted through the support 4 was used to determine the optical density. A signal may also be obtained. Alternatively, sample components may be fluorescently labeled and an optical density signal may be obtained using this fluorescent light.

Further, the rotating filter 7 may be arranged between the light source 5 and the support 4.

[Brief explanation of the drawing]

The drawings illustrate embodiments of the invention,
Fig. 1 is a schematic block diagram of a sample component quantification device showing an embodiment, Figs. 2 and 3 are diagrams showing the relationship between wavelength λ and transmittance T of different optical filters, and Figs. 4 a and b. 2A and 2B are schematic plan views showing other rotary filters, respectively. 1: Sample component quantitative device, 2: Optical system, 3: Signal processing system, 4: Support, 5: Light source, 7, 21, 2
2: Rotating filter, 8a, 8b, 8c, 8d: Optical filter, 11: Imaging device, 14: Switching device, 1
6: Computing device, 18: Monitor TV, 19: Printer.

Claims (1)

[Claims] 1. A sample, a support on which the sample is applied and separated into each component, and the sample components are dyed, an irradiation means for irradiating the support with light, and a support according to the wavelength of the light. a plurality of types of transmitting means having different transmittances; an imaging means for capturing an image of the light that is irradiated onto the support and transmitted through the transmitting means; and an image output of the imaging means based on the transmittance of the transmitting means. and a display means for displaying the output of the arithmetic means, and the transmitting means has an appropriate transmittance depending on the optical density of the light from the support. What is claimed is: 1. A sample component quantification device, wherein an image of transmitted light is captured with a constant sensitivity, and each component of the sample is imaged and quantified by an imaging means. 2. The sample component quantification device according to claim 1, wherein the imaging means is a single imaging device, and takes images of light in a time-divisional manner in response to switching of the transmission means. .