JPH0781946B2 - Fluorometer - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an improvement in a fluorometer, and more particularly to the structure of a sample cell section for supplying a sample for measurement.

[Prior Art] A conventional sample cell and sample supply device of a fluorometer have a structure as shown in FIG. 12, for example. The sample is introduced into the channel 2 in the sample cell 1 from the upper tube 9 through the channel 4 in the connecting portion 3 and the tapered portion 5, and is discharged from the lower tube.

FIG. 13 shows a sample chamber 300 provided in the middle of the sample cell 1 described above.
FIG. FIG. 13A shows the structure disclosed in Japanese Patent Laid-Open No. 52-136675, in which the sample portion 20 in the sample chamber 300 is surrounded by the transparent member 21 and the opaque member 22. The sample was irradiated with the excitation light 31 through the transparent member, and the fluorescence 32 emitted from the sample was taken out from the direction perpendicular to the incident angle of the excitation light.

FIG. 13 (b) shows the structure disclosed in Japanese Patent Laid-Open No. Sho 60-78334, in which the width of the transparent member on the incident side of excitation light is increased to increase the irradiation amount of excitation light to improve the sensitivity. The transparent member on the side opposite to the fluorescence emission side is replaced with an opaque member to remove scattered light from that portion and reduce the noise level.

[Problems to be Solved by the Invention] In the case of the sample supply device of FIG. 12 according to the above-described conventional technique, the taper portion 5 in the connecting portion 3 and the sample volume accommodated in the flow path 2 in the sample cell 1 are Since the sample volume in the channel 4 is too large, there is a problem that the band of fluorescence intensity spreads.
In the above-mentioned fluorescence measurement, a plurality of components in the sample solution are previously separated by a separation column, introduced sequentially into the sample cell, and the fluorescence intensity thereof is sequentially measured. At this time, the flow path 2 in the sample cell
If the volume in the connecting portion 3 is larger than the volume of the above, it takes time to replace the samples having different components, and the components spread in the connecting portion, so that the band expands.

Further, in the case of the sample cell of FIG. 13 (a), the irradiation amount of the excitation light 31 is limited by the width of the transparent member 21, and the scattered light from the transparent member on the side opposite to the fluorescence emission side is fluorescence 32. However, there is a problem that it is mixed in the inside and limits the sensitivity of the fluorescence measurement.

In the case of FIG. 13 (b), although the drawbacks of the structure of FIG. 13 (a) are improved, the mechanical strength is weakened because the adhesive area of the transparent member 211 on the fluorescence emission side is reduced, and the sample Department
If the cross-sectional area of 20 is made constant, the emission area of the fluorescence is reduced, so that there is a problem that the fluorescence emission amount is reduced. Usually, non-fluorescent transparent quartz is used for the transparent member, and black quartz is used for the opaque member. Of these, black quartz is fragile, and the sample chamber 300 has a minute structure, so a sufficient mechanical property is obtained. It was difficult to secure the physical strength.

It is an object of the present invention to provide a fluorometer capable of suppressing the band broadening, the increase of noise due to scattered light, etc., and further increasing the signal intensity, and at the same time obtaining a sufficient mechanical intensity.

[Means for Solving the Problems] In order to achieve the above-mentioned object, the present invention directly connects a tube 9 for introducing a sample to a flow path 2 of a sample cell 1 to eliminate unnecessary samples accumulated in a connecting portion 3. Try to eliminate it.

Further, the cross-sectional shape of the sample chamber in the sample cell 1 is made square to enhance the mechanical strength, and the opposite surface of the fluorescence emission surface is made of an opaque member to remove scattered light.

Further, the thickness of the opaque member provided in order to prevent the excitation light incident on the sample chamber from being mixed into the output fluorescence is reduced, so that the incident amount of the excitation light is increased.

Further, the sample chamber is configured by using four rectangular cross-section bodies in which an opaque member and a transparent member are bonded together, and the productivity, strength, etc. of the sample chamber are increased.

Further, the trapezoidal cross-section body is used to configure the sample chamber, three of the trapezoidal cross-section bodies are provided with an opaque body for covering the slope portion, and the other one of the constituent members is provided with the slope surface. An opaque body is provided to cover the portion and the top, so that the productivity and strength of the sample chamber are increased.

[Operation] The fluorescence photometer of the present invention configured as described above removes the sample pool at the connecting portion between the sample cell and the tube.

Further, the mechanical strength of the sample chamber in the sample cell is strengthened.

Furthermore, the amount of excitation light incident on the sample chamber, the amount of excitation light emitted therefrom, the amount of fluorescence emitted, and the like are increased, and at the same time unnecessary scattered light is excluded.

[Embodiment] An embodiment of the present invention will be described below with reference to the drawings. First
FIG. 2 and FIG. 2 are sectional views of the sample cell portion according to the present invention.

In FIG. 1, the connecting portion 3 of the conventional device shown in FIG.
The shape of the taper part 5 and the flow path 4 inside is changed, and the tube 9
Are directly connected to the flow path 2 of the sample cell 1. As a result, the sample accumulated in the connecting portion 3 can be eliminated, so that the band spread can be reduced. The ferrule 6 is fitted to the tube 9, and the ferrule 9 is pushed by the push screw 8 to push the tube 9 against the flow path 2 of the sample cell.

In FIG. 2 (a), a countersink 21 is provided at the end of the flow path 2 of the sample cell 1 in FIG. 1, and a tapered portion is provided at the end of the tube 9 to facilitate the mating of the two. It is the one. Further, FIG. 2B shows that the countersink portion is provided on the tube 9 side so that the tube 9 and the flow path 2 can be connected smoothly even if there is some positional deviation. In any case, it has an effect of eliminating the amount of the sample accumulated in the connecting portion 3.

FIG. 3 shows that the tube 9 having an inner diameter of 0.15φ of the sample supply device of FIG.
It is a measurement result of the band spread width in the case of μl. However, the sample is a 10ppm rhodamine B aqueous solution 0.1μ
1, the excitation light wavelength is 300 nm, and the fluorescence wavelength is 570 nm. From this, it can be seen that the band spread value decreases as the sample cell volume decreases.

On the other hand, in the same measurement by the conventional apparatus shown in FIG. 4, no significant difference was observed in the band spread with respect to the sample cell volume.

Therefore, according to the present invention, it is possible to reduce the sample cell volume and perform the measurement with a small band spread.

Next, an embodiment of the present invention relating to the sample chamber provided in the sample cell 1 will be described.

FIG. 5 is a sectional view of the sample chamber according to the present invention. Fifth
In the figure, since the opposite side of the emission surface of the fluorescence 32 of the conventional device shown in FIG. 13 (a) is an opaque member, there is no scattered light generated from this portion, and the transparent member on the fluorescence emission surface is thick. Therefore, sufficient mechanical strength can be obtained.

FIG. 6 is an assembly diagram of the sample chamber of the present invention. As shown in FIG. 7A, two transparent members 212 are placed on an opaque member 221, and opaque members 222 and 223 sandwiching the transparent member 213 are arranged on the transparent member 212. Bonding is performed by bonding as shown in FIG. 3B, and the sample portion 20 is formed at the center thereof. For example, non-fluorescent transparent quartz is used for each transparent member, and black quartz is used for the opaque member.

FIG. 7 shows the result of measurement of the amount of Raman scattering of water in the sample chamber of the present invention as compared with that of the conventional one. Excitation wavelength is 350nm, Raman peak wavelength is detected at 395nm,
The sample cell volumes were both 0.5 μl. 11A shows the case where the sample chamber of the present invention shown in FIGS. 5 and 6 is used, and FIG. 13B shows the case of the conventional apparatus shown in FIG. 13A.
Although the fluorescence intensity average values of the two are the same, the fluctuation (noise level) of the present invention is half or less, and it can be seen that the measurement sensitivity is more than doubled. This is because scattered light disappeared from the side opposite to the fluorescence emission surface.

FIG. 8 is a sectional view showing another sample chamber structure according to the present invention. In FIG. 8, consideration is given to increasing the pumping light 31 and the amount of the pumping light emitted from the pumping light 31, and at the same time obtaining sufficient mechanical strength. That is, comparing FIG. 8 with FIG. 5,
First, the opaque member 221 in FIG. 5 is thinned as in 224, transparent members 211 and 212 are extended in the upper and lower portions as in 214 and 215, and the opaque member 223 is thinned in as 225. A transparent member 216 is provided thereon. As a result, the incident area and the exit area of the excitation light increase, and
Since the bonding area between each member is the same as in the case of FIG. 5, equivalent mechanical strength can be obtained.

FIG. 9 shows the results of fluorescence measurement of the 0.1 ppm quinine sulfate aqueous solution in the sample chamber of FIG. 8 in comparison with the case of FIG. 13 (a). However, the sample cell capacity is 0.5μ
1, the excitation wavelength was 250 nm, and the measurement time constant was 300 ms. From this, in the case of FIG. 8, since the incident amount of the excitation light is increased,
As shown in A, it can be seen that the fluorescence intensity is significantly increased as compared with B in the case of FIG. 13 (a).

10 and 11 are views showing another structure of the sample chamber according to the present invention.

In Fig. 10, as shown in Fig. 10 (a), a rod body having a rectangular cross section in which an opaque member and a transparent member are pasted together is cut out, and four sticks are stuck together as shown in Fig. 10 (b). . C1 ~ C4
Etc. correspond to FIG. The excitation light 31 is incident from the transparent member portion of C1, the fluorescence 32 is emitted from the transparent member portion of C2, and the opposite surface is shielded by the opaque member of C4.
Further, the excitation light for monitoring is taken out from the transparent member portion of C3. The structure shown in FIG. 10 is characterized in that it is easy to manufacture and there is little variation in the finished sample chamber.

In FIG. 11, a bar body in which an opaque member is attached to the slope of the trapezoidal transparent member shown in FIG. 11A, and an opaque member on the slope and the top of the trapezoidal transparent member shown in FIG. Prepare stick-shaped and stick-shaped sticks, cut out each stick and stick them together as shown in FIG. D1 to D3 are those shown in FIG. 9A, and D4 is a member cut out from FIG. Excitation light
31 is incident from the transparent member portion of D1, fluorescence 32 is emitted from the transparent member portion of D2, and the opposite surface is shielded by the opaque member portion of D4. Further, the excitation light for monitoring is taken out from the transparent member portion of D3. The sample chamber in Figure 11 is easy to manufacture,
There is little variation in the finished product, and because the bonding surface of each member is long, strong adhesion can be obtained, and since the excitation light and fluorescence can have a wide incident angle and emission angle range, there is little loss of light. It has features such as high detection efficiency.

Further, in the structure of FIG. 11 (c), for example, D1 and D3 which do not have an opaque body on their slopes may be used.

[Advantages of the Invention] When the present invention is applied as described in detail above, the sample pool at the connecting portion between the sample cell and the tube can be removed,
The band spread of the fluorescence output can be narrowed, which enables ultrafast analysis of the sample.

Further, the bonding area between the transparent body and the opaque body used in the sample chamber in the sample cell can be increased to enhance its mechanical strength.

Furthermore, the excitation light incident amount to the sample chamber, the same emission amount, and the fluorescence emission amount, etc. are increased, and at the same time, the opposite side of the fluorescence emission surface is made opaque to remove unnecessary scattered light and reduce noise,
The sensitivity of the fluorometer can be increased.

[Brief description of drawings]

FIGS. 1, 2 (a) and 2 (b) are cross-sectional views showing the connection between the sample cell and the sample supply tube of the fluorometer according to the present invention, and FIG. 3 is the fluorescence band according to the device of the present invention. FIG. 4 shows the measurement result of the spread, FIG. 4 shows the measurement result of the band spread of the conventional device, FIG. 5 shows the sectional view of the sample chamber according to the present invention, and FIG. 6 (a) shows the sample chamber according to the present invention. Arrangement of transparent and opaque members used, FIG. 6 (b) is a perspective view of the sample chamber according to the present invention, FIG. 7 (a) is an output waveform diagram of the fluorometer according to the present invention, and FIG. 7 (b). FIG. 8 is an output waveform diagram of a conventional fluorometer, FIG. 8 is a cross-sectional view of another sample chamber according to the present invention, and FIG. 9 compares the fluorescence intensity measurement results of the fluorometer of the present invention and the conventional fluorometer. FIG. 10 (a) is a sectional view of a member used in another sample chamber of the present invention, FIG. 10 (b)
Is a cross-sectional view of the sample chamber of the present invention using the member of FIG.
11 (a) and 11 (b) are cross-sectional views of each member used in another sample chamber of the present invention, and FIG. 11 (c) uses the members shown in FIGS. 11 (a) and 11 (b). FIG. 12 is a sectional view of the sample chamber of the present invention, FIG. 12 is a sectional view showing the connection between the sample cell and the sample supply tube of the conventional fluorometer, and FIGS. 13 (a) and 13 (b) are conventional samples. It is sectional drawing of a chamber. 1 ... Sample cell, 2 ... Flow path, 3 ... Connection part, 5 ... Tapered part, 6 ... Ferrule, 7 ... Mounting screw, 8 ...
Push screw, 9 ... Tube, 11 ... Pedestal, 20 ... Sample part, 31 ... Excitation light, 32 ... Fluorescence, 21, 211 etc .... Transparent member, 22, 221 etc .... Opaque member, 300 ... … Sample chamber, C1 ……
Sample chamber components, D1 ... Sample chamber components.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-60-207038 (JP, A) JP-A-57-153248 (JP, A) JP-A- 64-91037 (JP, A) JP-A-1- 109245 (JP, A) JP 60-78334 (JP, A) JP 62-6142 (JP, A) Actually opened 53-142378 (JP, U) Actually opened 52-136675 (JP, U) Actual Kaihei 1-136449 (JP, U)

Claims (1)

[Claims] 1. A side surface of a sample chamber having a square cross section provided in a sample cell for introducing and discharging a sample by a tubular body is used as an excitation light incident surface, and is orthogonal to the excitation light incident surface of the sample chamber. In a fluorescence photometer for measuring the spectrum of fluorescence using one of the two side faces as a fluorescence emission surface, a transparent member having a width equal to that of the fluorescence emission surface is provided on the fluorescence emission surface. Each of the two side surfaces parallel to the excitation light incident surface is covered with an opaque member, the thickness of the opaque member on the excitation light incident surface side is made thinner than the other, and the opposite side of the fluorescence emission surface of the sample chamber. A fluorescent photometer, characterized in that another opaque member having the same width as the fluorescent emission surface is provided on the surface.