CN112461806B - Fluorescence spectrum detection method based on smart phone - Google Patents
Fluorescence spectrum detection method based on smart phone Download PDFInfo
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- CN112461806B CN112461806B CN202011295032.0A CN202011295032A CN112461806B CN 112461806 B CN112461806 B CN 112461806B CN 202011295032 A CN202011295032 A CN 202011295032A CN 112461806 B CN112461806 B CN 112461806B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
- G01N2021/6421—Measuring at two or more wavelengths
Abstract
A fluorescence spectrum detection method based on a smart phone relates to spectrum detection and image processing. 1) And (3) fluorescent signal detection: detecting a fluorescence signal by using a camera of the smart phone as a detector; 2) image data processing: processing video data shot by the smart phone; 3) drawing a spectral curve: selecting a proper color value as a fluorescence intensity parameter of the sample according to the color channel information of the effective luminescent area of the sample fluorescence image read in the step 2), and drawing a characteristic fluorescence spectrum curve of the sample by taking the wavelength as a horizontal coordinate and the fluorescence intensity as a vertical coordinate; and generating a characteristic fluorescence spectrum curve of the sample, defining a name by user, storing the name in a designated folder, and generating a fluorescent substance spectrum database. The fluorescent spectrum of a substance can be rapidly and accurately scanned in a video form through an image sensor of a camera of the smart phone; the method is simple and convenient to operate, short in analysis time consumption, and capable of improving detection sensitivity by reducing interference of environmental noise.
Description
Technical Field
The invention relates to the technical field of spectrum detection and image processing, in particular to a fluorescence spectrum detection method based on a smart phone.
Background
The fluorescence analysis method has the advantages of high sensitivity, strong selectivity, various methods and the like, and is widely applied to the fields of chemistry, biology, medicine, health, agriculture, environmental protection and the like. The fluorescence spectrum analysis method is used as a characterization technology for researching physical and chemical properties and change conditions of a system, and can be used as a means for qualitative detection and quantitative analysis. The traditional fluorescence analysis method is complex in operation, not beneficial to popularization, large in instrument size, high in manufacturing cost and difficult to apply to field detection. Therefore, the research of the novel rapid fluorescence analysis technology and the portable fluorescence analysis instrument has great significance and wide development prospect.
The smart phone is used as a simple and portable functional mobile device, and provides a new development opportunity for a portable detection technology. With the continuous perfection of the camera shooting function of the smart phone, the wide use of the APP and the continuous improvement of the data processing capacity, the smart phone can be used as an image acquisition tool and a digital processing device to play a strong role. The principle of the method is that optical signals, electric signals or color characteristics of a substance to be detected or a reaction process of the substance to be detected are collected in a picture mode, then the collected picture is subjected to signal quantization through special picture processing software or an APP installed on a smart phone, and finally a data result is obtained. The intelligent mobile phone is combined with the traditional detection technology, so that the advantages of lightness, portability, low cost, simplicity in operation, rapidness in processing and the like of the intelligent mobile phone can be fully exerted, the principle and the applicability of the traditional detection method are utilized, the defect of the traditional large instrument in the aspect of detection effectiveness can be overcome, and a huge development platform is provided for the research of the on-site rapid detection method.
The conventional fluorescence analyzer mainly uses a PMT (photomultiplier tube) as a photodetector, and needs to provide a stable negative high voltage device, which makes it difficult to miniaturize the analyzer. At present, an optical element is often required to be additionally arranged in a device adopted by a portable spectrum detection method based on a smart phone, the light path design is complex, and the cost is high. Meanwhile, the existing spectrum detection method based on the smart phone obtains the spectrum by taking a single sample picture, and has the defects of small detection range, single detection object, complicated operation in the image processing process, long analysis time and the like. Therefore, the development of the fluorescence spectrum detection method based on the smart phone, which is simple in structure, convenient and fast to operate and wide in application range, has great application value.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, and provides the fluorescence spectrum detection method based on the smart phone, which can be widely applied to the fluorescence spectrum detection of fluorescent substances in a visible light range, can be adapted to various types of smart phones, and has the advantages of simple structure, convenience in operation and high detection accuracy.
The invention comprises the following steps:
1) and (3) fluorescent signal detection: detecting a fluorescence signal by using a camera of the smart phone as a detector;
2) image data processing: processing video data shot by the smart phone;
3) drawing a spectral curve: selecting a proper color value as a fluorescence intensity parameter of the sample according to the color channel information of the effective luminescent area of the sample fluorescence image read in the step 2), and drawing a characteristic fluorescence spectrum curve of the sample by taking the wavelength as a horizontal coordinate and the fluorescence intensity as a vertical coordinate; and generating a characteristic fluorescence spectrum curve of the sample, defining a name by user, storing the name in a designated folder, and generating a fluorescent substance spectrum database.
In step 1), the specific steps of detecting the fluorescence signal by using the camera of the smartphone as the detector may be: aligning a rear camera of the smart phone to a slit of a monochromator, opening a camera of the smart phone, detecting a fluorescent signal transmitted from the slit of a light outlet of the monochromator by using an image sensor of the smart phone, and recording the fluorescent signal transmitted from a sample irradiated by an excitation light source, which is subjected to light splitting by the monochromator and transmitted from the slit, in a video shooting mode by using a video recording function of the camera of the smart phone;
the intelligent mobile phone can adopt any type of mobile phone with a camera, and an image sensor of the camera of the intelligent mobile phone is used as a detection element; the rear camera of the smart phone is aligned to the slit of the monochromator, the smart phone can be fixedly placed on the mobile phone support firstly, the camera of the smart phone is aligned to the slit of the light outlet of the monochromator, and the detector of the smart phone, the monochromator and the sample are positioned on the same horizontal straight line and are arranged at a 90-degree right angle with the light source.
In step 2), the specific steps of processing the video data obtained by shooting with the smartphone may be: matching the duration of the video data with the wavelength range of the fluorescence emission of the sample, converting the number of fluorescence images according to the ratio of the wavelength range of the fluorescence emission of the sample to the spectrum scanning step length, and converting the video data into a series of sample fluorescence images corresponding to the wavelength; performing rapid batch processing on the series of sample fluorescence image data, including selecting an effective light emitting area of the sample fluorescence image, and reading different color channel information of the area;
in step 2), the specific method for performing rapid batch processing on the fluorescence image data may be: selecting the size of an effective light-emitting area of a fluorescence image of a sample according to actual light-emitting areas of different samples, storing the shape of a frame line of the selected area after the selection is finished, and automatically selecting the same light-emitting area of the same group of fluorescence image data in batches in a follow-up manner; when the fluorescence signal of the sample is weak or the interference signals such as environmental noise and the like are strong, image noise reduction treatment can be firstly carried out;
and reading the different color channel information of the area is to read the different color channel information of the effective luminescent area of the sample fluorescence image, and the obtained result of the color value of each channel is the average value of the color values of the channel in the area.
In step 3), the appropriate color value is selected according to the actual fluorescence emission wavelength region of different fluorescent substances so as to reflect the real fluorescence emission spectrum information of the fluorescent substances.
Compared with the prior art, the invention has the beneficial effects that:
the invention designs a novel fluorescence spectrum detection method based on a smart phone, which realizes that the fluorescence spectrum of a substance is rapidly and accurately scanned in a video form through an image sensor of a camera of the smart phone; the method has the advantages that video data shot by the smart phone are converted into sample fluorescence images, the image data are rapidly processed in batches, the operation is simple and convenient, the analysis time is short, and the detection sensitivity can be improved by reducing the interference of environmental noise. The obtained experimental result has higher matching degree compared with the substance fluorescence spectrum scanning result obtained by the conventional fluorescence spectrophotometer. The invention adopts the image sensor of the smart phone to replace a photomultiplier used by a conventional fluorescence spectrophotometer as a detection element of a fluorescence signal, simulates the spectrum scanning mode of a photoelectric detector, adopts a smart phone camera to continuously collect the fluorescence signal of a sample in a video shooting mode, converts the light signal of the sample into color channel information of the image, directly utilizes color values as parameters, can obtain the fluorescence spectrum of the sample, and can realize the fluorescence spectrum detection of different fluorescent substances in a larger wavelength range. The miniaturized monochromator and the excitation light sources with different wavelengths are matched, and an additional optical element and a complex light path arrangement are not needed, so that the cost is saved, and the volume of the instrument is reduced. Meanwhile, the adjustable and movable smart phone support is arranged on the fluorescence signal detection part of the system, smart phones of different models can be adapted to the system to serve as fluorescence signal detectors, and accurate and rapid fluorescence spectrum detection of different types of smart phones for different fluorescent substances can be guaranteed.
Drawings
Fig. 1 is a general flow diagram of the present invention.
FIG. 2 is a schematic flow chart of a sample fluorescence image data processing method according to the present invention.
FIG. 3 is a diagram illustrating the effective light-emitting area of the fluorescence image selected during the fluorescence image data processing process of the sample according to the present invention.
FIG. 4 is a substance fluorescence spectrum characterized by different color values during the process of drawing the sample characteristic spectrum curve according to the present invention.
FIG. 5 shows fluorescence emission spectra of 1mg/L dansyl chloride solution detected using the present invention and its comparison with conventional fluorescence spectrophotometer detection results.
FIG. 6 is a fluorescence emission spectrum of 5mmol/L rhodamine 6G solution detected by the method and a comparison of the fluorescence emission spectrum and a detection result of a conventional fluorescence spectrophotometer.
Detailed Description
The following examples will further illustrate the present invention with reference to the accompanying drawings.
As shown in fig. 1, a fluorescence spectrum detection method based on a smart phone includes the following steps:
placing the smart phone on a mobile phone support, and enabling a rear camera of the smart phone to be aligned with a slit of a monochromator; and opening a camera of the smart phone, detecting a fluorescent signal transmitted from a slit of a light outlet of the monochromator by using an image sensor of the smart phone, clicking a video recording function of the camera of the smart phone, and recording the fluorescent signal transmitted by the sample within a certain wavelength range in a video shooting mode, wherein the fluorescent signal is emitted by the sample after the sample is irradiated by an excitation light source and passes through the monochromator.
As shown in fig. 2, the sample fluorescence image data processing specifically includes the following steps:
processing video data obtained by shooting of a smart phone, firstly matching the duration of the video data with the wavelength range of sample fluorescence emission, converting the number of fluorescence images according to the ratio of the wavelength range of the sample fluorescence emission to the spectrum scanning step length, and converting the video data into a series of sample fluorescence images corresponding to the wavelength; then, batch processing is carried out on the series of sample fluorescence images, wherein the batch processing comprises selecting an effective light emitting area of image data, as shown in FIG. 3; reading different color channel information of an effective light emitting area of the image; selecting a proper color reference value to represent the fluorescence intensity of the sample, and drawing a sample characteristic fluorescence spectrum curve by taking the wavelength as an abscissa and the fluorescence intensity as an ordinate, as shown in FIG. 4. And storing the obtained characteristic fluorescence spectrum curve of the sample in a designated folder, and finally generating a spectrum database.
The present invention was tested with dansyl chloride solution and rhodamine 6G solution as examples, as shown in fig. 5 and 6, with the abscissa being wavelength (nm) and the ordinate being normalized fluorescence intensity; the left panel shows the fluorescence emission spectra detected using a conventional spectrofluorometer and the right panel shows the fluorescence emission spectra detected using the present invention. As shown in FIG. 5, the position of the fluorescence emission peak of the dansyl chloride solution obtained by scanning with a conventional fluorescence spectrophotometer is 526nm, the position of the fluorescence emission peak of the dansyl chloride solution obtained by scanning with the present invention is 527nm, and the fluorescence emission spectra of the dansyl chloride solutions obtained by scanning with the two methods are substantially consistent; as shown in FIG. 6, the fluorescence emission peak position of the rhodamine 6G solution scanned by using a conventional fluorescence spectrophotometer is at 577nm, the fluorescence emission peak position of the rhodamine 6G solution scanned by using the method is at 577nm, and the fluorescence emission spectra of the rhodamine 6G solution scanned by the two methods are basically consistent. The experimental result shows that the method has reliability.
The invention uses the camera of the smart phone to replace a photomultiplier tube used by the traditional fluorescence spectrophotometer to detect the fluorescence signal of the sample; and converting video data shot by the smart phone into a sample fluorescence image, and rapidly processing the image data in batches. Selecting an effective light emitting area of a sample fluorescence image, and reading channel information of different colors in the area; and selecting a proper color value as a fluorescence intensity parameter of the sample, and drawing a characteristic fluorescence spectrum curve of the sample by taking the wavelength as an abscissa and the fluorescence intensity as an ordinate. The invention is widely suitable for the fluorescence spectrum detection of the fluorescent substance in the visible light range, can be adapted to various types of smart phones, and has the advantages of simple structure, convenient operation and high detection accuracy.
Claims (6)
1. The fluorescence spectrum detection method based on the smart phone is characterized by comprising the following steps of:
1) and (3) fluorescent signal detection: detecting a fluorescence signal by using a camera of the smart phone as a detector;
2) image data processing: processing video data shot by the smart phone;
the specific steps of processing the video data shot by the smart phone are as follows: matching the duration of the video data with the wavelength range of the fluorescence emission of the sample, converting the number of fluorescence images according to the ratio of the wavelength range of the fluorescence emission of the sample to the spectrum scanning step length, and converting the video data into a series of sample fluorescence images corresponding to the wavelength; performing rapid batch processing on the series of sample fluorescence image data, including selecting an effective light emitting area of the sample fluorescence image, and reading different color channel information of the area;
3) drawing a spectral curve: selecting a proper color value as a fluorescence intensity parameter of the sample according to the color channel information of the effective luminescent area of the sample fluorescence image read in the step 2), and drawing a characteristic fluorescence spectrum curve of the sample by taking the wavelength as a horizontal coordinate and the fluorescence intensity as a vertical coordinate; and generating a characteristic fluorescence spectrum curve of the sample, defining a name by user, storing the name in a designated folder, and generating a fluorescent substance spectrum database.
2. The method for detecting fluorescence spectrum based on smart phone according to claim 1, wherein in step 1), the specific steps of using the camera of the smart phone as the detector to detect the fluorescence signal are as follows: the method comprises the steps of aligning a rear camera of the smart phone to a slit of a monochromator, opening a camera of the smart phone, detecting a fluorescent signal transmitted from the slit of a light outlet of the monochromator by using an image sensor of the smart phone, and recording the fluorescent signal transmitted from a sample after being irradiated by an excitation light source, which is split by the monochromator and transmitted from the slit, in a video shooting mode by using a video recording function of the camera of the smart phone.
3. The fluorescence spectrum detection method based on the smart phone as claimed in claim 2, wherein in step 1), the smart phone adopts any type of mobile phone with a camera, and an image sensor of the camera of the smart phone is used as a detection element; the rear camera of the smart phone is aligned to the slit of the monochromator, the smart phone is fixedly placed on the mobile phone support firstly, the camera of the smart phone is aligned to the slit of the light outlet of the monochromator, and the detector of the smart phone, the monochromator and the sample are positioned on the same horizontal straight line and are arranged at a 90-degree right angle with the light source.
4. The fluorescence spectrum detection method based on the smart phone as claimed in claim 1, wherein in step 2), the specific method for performing the rapid batch processing on the series of sample fluorescence image data is as follows: selecting the size of an effective light-emitting area of a fluorescence image of a sample according to actual light-emitting areas of different samples, storing the shape of a frame line of the selected area after the selection is finished, and automatically selecting the same light-emitting area of the same group of fluorescence image data in batches in a follow-up manner; and when the fluorescence signal of the sample is weaker or the environmental noise interference signal is stronger, image noise reduction processing is performed firstly.
5. The method according to claim 1, wherein in step 2), the reading of the different color channel information of the area is reading of the different color channel information of an effective light emitting area of the sample fluorescence image, and the obtained result of each channel color value is an average value of the channel color values in the area.
6. The method as claimed in claim 1, wherein in step 3), the appropriate color value is selected according to the actual fluorescence emission wavelength region of different fluorescent substances to reflect the real fluorescence emission spectrum information of the fluorescent substances.
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| CN113340858B (en) * | 2021-04-30 | 2023-02-03 | 南京理工大学 | Portable mixed fluorescence detector |
| CN117074321A (en) * | 2023-08-15 | 2023-11-17 | 浙江大学 | Method for detecting chemical components of extracting solution based on infrared light information smart phone |
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