Optical device for performing fluorescence and absorbance measurements

Abstract

The invention relates to an optical device configured for measuring fluorescence and absorbance in immunoassays and wet chemistry, comprising:-an illuminator channel (26) comprising at least one excitation source (28),-a detector channel (34) comprising at least one sensor (36),-a cuvette (130) comprising a sample (132) between the illuminator channel (26) and the detector channel (34), -an optical light mixer (38) between the illuminator channel (26) and the cuvette (130) and an optical splitter (40) between the cuvette (130) and the detector channel (34).

Description

Technical Field

[0001] This disclosure relates to the field of absorbance and fluorescence measurement using optical devices. Background Technology

[0002] This application relates to the field of absorbance and fluorescence measurement.

[0003] In fluorescence measurements, the sample is illuminated by an illumination source at a precise excitation wavelength (e.g., 370 nm). The sample then emits a different wavelength than the excitation wavelength, e.g., 450 nm.

[0004] In absorbance measurement, the attenuation of light due to the presence of a sample is measured. First, a cuvette without a sample is set up. An illumination source is turned on, and the received light is measured after passing through an empty or water-filled cuvette. Then, the sample is placed in the cuvette, and the illumination source is turned on again. The attenuation of light in the presence of the sample is measured. More precisely, the absorbance of the sample is obtained by dividing the decimal logarithm of the light measured with the empty or water-filled cuvette by the light measured with the sample.

[0005] In existing technologies, such as Figure 1 As shown, a motorized optical device 2 exists for measuring absorbance and fluorescence. This motorized optical device 2 consists of an emission channel 4 with a white light 6 and a first lens 8, a first motorized filter wheel 10 and a second motorized filter wheel 12, a sample 14 in a cuvette 16, and a detection channel 18. The first motorized filter wheel 10 is moved to select a wavelength from the white light 6 for illuminating the sample 14. Simultaneously, in the detection channel 18, the second motorized filter wheel 12 is used to select a wavelength for detection. In this case, a broadband illumination source and a broadband detector 20 with a second lens 22 are used. Wavelength selection is accomplished by these first motorized filters 10 and second motorized filters 12.

[0006] Therefore, there are two motor components that exert mechanical constraints on the device. Furthermore, the size of each motorized filter wheel is significant. This significant size of the first motorized filter wheel 10 and the second motorized filter wheel 12 results in a large footprint in the instrument in which these motorized filters are implemented. In fact, the arrangement in the prior art is not optimal because the cuvette is positioned between the first motorized filter wheel 10 and the second motorized filter wheel 12, thus the two motorized components are independent and separated from each other, which multiplies the number of components within the device and complicates the device. Reliability issues also arise due to the moving parts of the motorized filters. Furthermore, alignment problems occur during the rotation of the motorized filters. Summary of the Invention

[0007] This disclosure improves upon this situation.

[0008] An analytical system is proposed, comprising optical devices configured to measure fluorescence and absorbance in a biological sample contained in at least one cuvette, the analytical system including: - An illuminator channel, the illuminator channel comprising at least one light source for each desired spectrum, - Detector channel, which includes at least one sensor for measuring the fluorescence and absorbance in the biological sample. - An optical mixer between the illuminator channel and the cuvette, and a beam splitter between the cuvette and the detector channel. - At least one cuvette configured to contain the biological sample, the cuvette being arranged between the illuminator channel and the detector channel. Because of this arrangement, the optical devices are configured to evaluate fluorescence and absorbance measurements in parallel and preferably simultaneously and independently, but using the same equipment, which allows for complex and precise data analysis of the samples.

[0009] The detector channel is configured to filter light as needed without the need for moving parts. The optical device is more stable than existing optical devices.

[0010] The optical devices are configured to be miniaturized and can be integrated into measuring instruments.

[0011] Optical equipment is particularly relevant for immunoassays by measuring fluorescence and absorbance in parallel and preferably simultaneously.

[0012] According to the present invention, "parallelism" means using the same equipment in the same analysis cycle.

[0013] An optical mixer can be an integrated combination of lenses and glass, configured to transmit a collimated single output beam from the illuminator channel to the cuvette.

[0014] This optical mixer consists of a small, stable component that does not move and is positioned within a molded housing without special alignment. The optical mixer allows simultaneous operation at different wavelengths and the use of different source driving and detection techniques. The glass of the optical mixer may include filters and at least one beam mixer.

[0015] A beam splitter can be an integrated combination of lenses and glass, configured to separate a single output beam that has passed through a cuvette into at least two detection beams.

[0016] The beam splitter consists of a small, stable component that does not move and is positioned within a molded housing without special alignment. This beam splitter allows simultaneous operation at different wavelengths and the use of different source driving and detection techniques. The glass of the beam splitter may include filters and at least one beam splitter.

[0017] The optical light mixer may include a first tricolor prism configured to transmit a collimated single output beam from the illuminator channel to the cuvette.

[0018] The first trichromatic prism is configured to avoid energy loss and obtain a collimated single output beam by selecting a specific wavelength.

[0019] The illuminator channel may include a first light source, a second light source, and a third light source for the spectrum.

[0020] The first light source used for the spectrum can be a white LED.

[0021] The second light source for the spectrum can be a UV LED configured to emit at 340 nm.

[0022] The third light source for the spectrum can be a UV LED configured to emit at 370 nm.

[0023] The beam splitter may include a second trichromatic prism configured to separate a single output beam that has passed through a cuvette into a first detection beam, a second detection beam, and a third detection beam.

[0024] The detector channel may include a first, second, and third sensor.

[0025] The first sensor may be a broadband sensor configured to detect wavelengths between 340 nm and 700 nm.

[0026] The second sensor can be a sensitivity sensor configured to detect a wavelength of 450 nm.

[0027] The second sensor is configured to detect very low signals.

[0028] The third sensor can be a sensor configured to detect a wavelength of 450 nm.

[0029] The analytical system includes analytical consumables, which include at least one cuvette configured to hold a biological sample.

[0030] The analytical consumables include a disposable body configured to cover and protect a radially shaped strip, wherein at least one cuvette is arranged within the radially shaped strip.

[0031] Advantageously, the radially shaped strips comprise multiple cuvettes.

[0032] The radially shaped strip is rotatably mounted in the optical device.

[0033] Advantageously, at least one other cuvette is pre-filled with reagent.

[0034] The analysis system includes an analysis unit configured to analyze signals from optical devices.

[0035] The analysis system includes at least one alarm device, which is configured to at least communicate the results of the analysis to the user. Attached Figure Description

[0036] Other features, details, and advantages will be shown in the following detailed description and accompanying drawings, wherein: Figure 1 A schematic diagram showing existing optical devices.

[0037] Figure 2 A three-dimensional view of a portion of an optical device according to the invention is shown, comprising radially shaped stripes that open in “part A” and close in “part B”.

[0038] Figure 3 A schematic diagram illustrating the optical device according to the invention integrated in a radially shaped strip.

[0039] Figure 4 express Figure 3 A schematic diagram of the illuminator channel and detector channel of the optical device shown.

[0040] Figure 5 express Figure 3 A schematic diagram of an optical device, with detailed views of the illuminator channel and detector channel in conjunction with... Figure 4 The opposite side compared to.

[0041] Figure 6 This indicates that according to the invention, it includes an optical guide. Figure 4 A schematic diagram of the optical device shown.

[0042] Figure 7 This diagram illustrates an optical mixer and beam splitter implemented using a tricolor prism according to the present invention.

[0043] Figure 8 A three-dimensional view of an optical device.

[0044] Figure 9 A schematic diagram showing a miniature spectrometer that functions as a sensor in an optical device.

[0045] Figure 10 A three-dimensional view of the cuvette of the present invention having a collimated single output beam.

[0046] Figure 11 This is a graph showing a preliminary comparison between the results obtained using the current fluorescence reading device and the results obtained using the optical device of the present invention.

[0047] Figure 12 It is a graph representing the absorbance test.

[0048] Figure 13 This is a perspective view of an embodiment of the present invention. Detailed Implementation

[0049] This invention relates to an optical device configured to measure the fluorescence and absorbance of small-volume samples in immunoassays and / or wet chemistry.

[0050] The present invention relates to an analytical system comprising an analytical consumable 100 having at least one cuvette 130 and an optical device cooperating with said at least one cuvette 130.

[0051] refer to Figures 2 to 6 The optical device of the present invention is configured to cooperate with analytical consumable 100, which includes at least one cuvette 130 configured to contain a biological sample 132. Figure 2 As can be seen, the analytical consumable 100 includes a disposable body 110 configured to cover and protect a radially shaped strip 120 in which at least one cuvette 130 is disposed. Furthermore, the radially shaped strip 120 includes a plurality of cuvettes 130. The radially shaped strip 130 is rotatably mounted in the optical device of the present invention.

[0052] A rotating four-channel end 140 is disposed on top of the disposable body 110: for example, three channels can be dedicated to sample pretreatment, and one channel can be docked with a pump. A determination label 150 is disposed on the rotating four-channel end 140, and a strip label 134 is disposed on a radially shaped strip 120.

[0053] The optical apparatus also includes an illuminator channel 26, a cuvette 130, and a detector channel 34. The illuminator channel 26 includes at least one light source 28 for each desired spectrum, and the cuvette 130 contains the sample 132 to be analyzed. The illuminator channel 26 generates multi-wavelength light. The detector channel 34 is configured to detect the multi-wavelength light. The detector channel 34 includes at least one sensor 36. The sensor 36 is configured to receive multi-wavelength light that has passed through the cuvette 130 or is emitted by the sample 132 within the cuvette 130. The optical apparatus is configured to evaluate fluorescence and absorbance measurements in parallel.

[0054] like Figure 4 , 5 As shown in Figure 6, illuminator channel 26 and detector channel 34 are interchangeable. Illuminator channel 26 and detector channel 34 do not have specific positions. This interchangeability provides flexibility in the positioning of illuminator channel 26 and detector channel 34.

[0055] according to Figure 7 An optical mixer 38 is positioned between the illuminator channel 26 and the cuvette 130. A spectrometer 40 is positioned between the cuvette 130 and the detector channel 34. At least one light source 28 for the spectrum is modulated by a suitable firmware (FW) and hardware (HW) system connected to at least one light source and receiver. These modulator and demodulator blocks are configured to modulate the light and eliminate all noise or electronic noise caused by ambient light.

[0056] The optical mixer 38 includes at least a first trichromatic prism 42. The first trichromatic prism 42 is configured to avoid energy loss and obtain a collimated single output beam 44 by selecting a specific wavelength. (Reference) Figure 6 and Figure 7 The first tricolor prism 42 allows the use of at least one light guide 46.

[0057] refer to Figure 7 and Figure 8 The illuminator channel 26 may include three light sources for spectroscopy. The first excitation source 48 is a white light-emitting diode (LED) with a broadband excitation source for spectroscopy. The term "broadband" means including wavelengths, for example, between 340 nm and 700 nm. This broadband excitation source for spectroscopy can be used for absorbance testing. The second excitation source 50 for spectroscopy is, for example, a UV LED configured to emit at 340 nm, and the third excitation source 52 for spectroscopy is, for example, an ultraviolet (UV) LED configured to emit at 370 nm. Each excitation source for spectroscopy emits a beam in its own wavelength band. The first tricolor prism 42 combines the three different wavelengths into a single output beam 44, which illuminates one side of the cuvette 130 containing the sample 132 for analysis. The first tricolor prism 42 is configured to obtain a collimated beam. The optical apparatus also includes a housing 54 for the cuvette 130, positioned between the illuminator channel 26 and the detector channel 34.

[0058] like Figure 7 As shown, on the detector channel 34 side, the wavelength to be detected is selected. The beam splitter 40 may include a second trichromatic prism 43. This second trichromatic prism separates the single output beam 44 into a first detection beam 56, a second detection beam 58, and a third detection beam 60. Due to the three sensors, the first detection beam 56, the second detection beam 58, and the third detection beam 60 are detected. For example, the first sensor 62 may be a broadband sensor that detects wavelengths including those between 340 nm and 700 nm. Figure 9 As shown, the first sensor 62 can be a miniature spectrometer. The selection of the wavelength to be measured is done within the miniature spectrometer itself. Inside the miniature spectrometer, there is an array of pixels 64 capable of receiving each specific wavelength 66 (see [link to image]). Figure 9Each wavelength of the detection beam illuminates a single pixel within the miniature spectrometer. The second sensor 68 can be a sensitivity sensor for detecting a wavelength of 450 nm. This second sensor 68 is configured to detect very low signals. A cuvette 130 containing a small amount of sample 130 requires detection of very low signals by the second sensor 68, especially for fluorescence measurements. The third sensor 70 can be a sensor for detecting a wavelength of 450 nm. The detector channel 34 is configured to filter light as needed without the need for moving parts. This optical device is more stable than existing optical devices.

[0059] The optical equipment uses the same cuvette 130 to perform fluorescence and / or absorbance measurements. Fluorescence and absorbance measurements can be performed simultaneously and / or in parallel, therefore, there is no crosstalk between the effects of the two tests. The cuvette 130 is designed with specific materials made from different types of plastics. For example... Figure 10 As shown, the shape of this cuvette 130 is also special because the sample volume is very small, typically ranging from 130 μL to 200 μL. The illumination volume is maximized, and the dead volume is minimized.

[0060] The optical device is configured for miniaturization and integration into measuring instruments. It has no moving parts and is therefore stable. The optical device complies with all mechanical constraints.

[0061] Optical equipment is particularly relevant for immunoassays and / or wet chemistry by allowing parallel measurements of fluorescence and absorbance. For example, procalcitonin can be measured by fluorescence, and creatinine can be measured by absorbance in parallel using the same equipment during the same cycle. These measurements are useful for assessing organ failure of the liver during sepsis.

[0062] Experimental Test refer to Figure 8Fluorescence and absorbance measurements have been performed. Illuminator channel 26 includes three excitation sources for spectroscopy. The first excitation source 48 for spectroscopy is a white LED with a broadband excitation source. This broadband excitation source can be used for absorbance measurement. The second excitation source 50 for spectroscopy is a UV LED configured to emit at 340 nm, and the third excitation source 52 for spectroscopy is a UV LED configured to emit at 370 nm. The optical apparatus also includes a housing 54 for the cuvette, positioned between illuminator channel 26 and detector channel 34. Detector channel 34 includes a microspectrometer corresponding to a broadband sensor detecting wavelengths between 340 and 700 nm. An optical mixer 38 is positioned between illuminator channel 26 and cuvette 130. This optical mixer 38 is a first tricolor prism 42. A spectrometer 40 is positioned between cuvette 130 and detector channel 34. A single broadband sensor, such as a microspectrometer, is positioned between cuvette 130 and detector channel 34.

[0063] A preliminary comparison between the results obtained using current fluorescence reading devices and the results obtained using the optical device of the present invention is given below. Figure 11 The graph in the figure represents fluorescence as pure counts on the vertical axis 72 and fluorophore concentration as "nM" on the horizontal axis 74. The solid line 76 represents the results obtained using the optical equipment of this invention, and the dashed line 78 represents the results obtained using current fluorescence measurement instruments. The small differences observed may be due to different settings of the excitation source for the spectrum and can be easily superimposed using a linear conversion factor.

[0064] It also achieved absorbance testing. Figure 12 The graph shows absorbance 80 on the vertical axis (82) and sample concentration in nM on the horizontal axis (84). Solid line 86 represents the result obtained using the optical device of this invention, and dashed line 88 represents the result obtained using a spectrophotometer. The same sample was used. The results obtained using the optical device of this invention are comparable to those obtained using a practical absorbance instrument.

[0065] exist Figure 13In the illustrated embodiment, illuminator channel 26 includes two sources: a UV (ultraviolet) LED configured to emit at 370 nm and a white LED configured to emit between 400 nm and 700 nm. UV LED 82 is configured to measure fluorescence and white LED 84 is configured to measure absorbance. The two LEDs can be used simultaneously and each modulated at a different frequency. Detector channel 34 includes two sensors: a multi-pixel photon counter (MPPC) sensor 86 configured to measure low fluorescence signals; and a spectrometer 88 configured to measure high-to-medium fluorescence signals and absorbance. The two sensors can be read simultaneously. Due to the different frequencies used to modulate the two excitation sources (UV LED and white LED), the spectrometer can distinguish between fluorescence and absorption light.

[0066] The optical light mixer 38 is an integrated combination of lenses and glass configured to transmit a collimated single output beam 44 from the illuminator channel 26 to the cuvette 130. The optical light mixer 38 includes a first lens 90 and a second lens 92, a first reflector 94 configured to refract light emitted by a UV LED, a first beam mixer 96, and a third lens 98. Between the optical light mixer 38 and the cuvette 130, the collimated single output beam 44 passes through a second reflector 204, a lens 206, a filter 208, a subsequent lens 210, and a third reflector 212.

[0067] Beam splitter 40 is an integrated combination of lenses and glass, configured to separate a single output beam 44 that has passed through a cuvette into at least two detection beams 80. Beam splitter 40 includes at least two lenses 200, two filters, and beam splitter 202.

Claims

1. Optical devices configured to measure fluorescence and absorbance in immunoassay and / or wet chemical samples, including: - Illuminator channel (26), which includes at least one light source (28) for each desired spectrum. - Detector channel (34), which includes at least one sensor (36) for measuring the fluorescence and absorbance in the immunoassay and / or wet chemical sample. - A cuvette (130) configured to contain the immunoassay or wet chemical sample (132) between an illuminator channel (26) and a detector channel (34). - An optical mixer (38) between the illuminator channel (26) and the cuvette (130) and a spectrometer (40) between the cuvette (130) and the detector channel (34).

2. The optical device according to claim 1, wherein the optical mixer (38) is an integrated combination of lens and glass configured to transmit a collimated single output beam (44) from the illuminator channel to the cuvette.

3. The optical device according to any one of claims 3 to 7, wherein the beam splitter (40) is an integrated combination of lens and glass configured to split the single output beam (44) that has passed through the cuvette into at least two detection beams 80.

4. The optical device according to claim 1, wherein the optical light mixer (38) includes a first tricolor prism (42) configured to transmit a collimated single output beam (44) from the illuminator channel to the cuvette.

5. The optical device according to claim 4, wherein the illuminator channel (26) comprises a first light source, a second light source and a third light source for the spectrum.

6. The optical device according to claim 5, wherein the first excitation source (48) for the spectrum is a white LED.

7. The optical device according to claim 5 or 6, wherein the second excitation source (50) for the spectrum is a UV LED configured to emit at 340 nm.

8. The optical device according to any one of claims 5 to 7, wherein the third excitation source (52) for the spectrum is a UV LED configured to emit at 370 nm.

9. The optical device according to any one of claims 4 to 8, wherein the beam splitter (40) comprises a second tricolor prism (43) configured to split the single output beam (44) that has passed through the cuvette into a first detection beam (56), a second detection beam (58) and a third detection beam (60).

10. The optical device according to claim 7, wherein the detector channel (34) comprises a first sensor (62), a second sensor (68) and a third sensor (70).

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