CN105241866A - Method for eliminating base line interference in Raman spectrum by utilizing fluorescent bleaching effect - Google Patents
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Abstract
一种利用荧光褪色效应消除拉曼光谱中基线干扰的方法，涉及一种消除拉曼光谱中基线干扰的方法，该方法是通过测量样品的光谱信号S，计算系列光谱信号的差值ΔS及其平均值ΔS’，并对差值平均值ΔS’作降噪处理，经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs；从样品光谱信号中逐步扣减荧光的度量值Fs，判断扣减是否达到要求，如果是，则消除了拉曼光谱中基线干扰，得到纯净的拉曼光谱信号。本发明能保证数据处理的真实性和合理性，所用的设备简单、造价低，方法简单，易于普及。
A method for eliminating baseline interference in Raman spectra by using fluorescence fading effect, relates to a method for eliminating baseline interference in Raman spectra, the method is by measuring the spectral signal S of a sample, calculating the difference ΔS of a series of spectral signals and its The average value ΔS', and the noise reduction process is performed on the difference average value ΔS', and the difference value average value ΔS' after noise reduction is used as the measurement value Fs of the fluorescence contained in the sample spectral signal; the fluorescence is gradually subtracted from the sample spectral signal The metric value Fs of , judges whether the subtraction meets the requirements, if so, eliminates the baseline interference in the Raman spectrum, and obtains a pure Raman spectrum signal. The invention can ensure the authenticity and rationality of data processing, the equipment used is simple, the cost is low, the method is simple, and it is easy to popularize.
Description
技术领域 technical field
本发明涉及一种消除拉曼光谱中基线干扰的方法，特别是一种利用荧光褪色效应消除拉曼光谱中基线干扰的方法。 The invention relates to a method for eliminating baseline interference in Raman spectrum, in particular to a method for eliminating baseline interference in Raman spectrum by utilizing the fluorescence fading effect .
背景技术 Background technique
激光拉曼光谱，目前已成为物质快速定性的重要方法。拉曼光谱由单波长激光激发产生，但是同时激发的荧光，对拉曼光谱的可辨识性带来很大干扰，是影响拉曼信号质量最主要的因素。尤其是近年来得到普遍应用的微型分光拉曼光谱仪，由于结构简单、便携、无需复杂维护，在现场测量中得到广泛应用；但由于是直接分光采集光谱，受到荧光干扰后，难以通过硬件加以消除；虽然更长的激发波长，可以降低荧光产生几率，但目前受到造价和元器件性能的约束；同时，更长波长激发效率低，烧毁样品的可能性也提高了。 Laser Raman spectroscopy has become an important method for rapid characterization of substances. The Raman spectrum is generated by excitation of a single-wavelength laser, but the fluorescence excited at the same time greatly interferes with the identifiability of the Raman spectrum, which is the most important factor affecting the quality of the Raman signal. In particular, the miniature spectroscopic Raman spectrometer, which has been widely used in recent years, has been widely used in on-site measurement due to its simple structure, portability, and no need for complicated maintenance. However, due to the direct spectroscopic collection of spectra, it is difficult to eliminate through hardware after being interfered by fluorescence. ; Although the longer excitation wavelength can reduce the probability of fluorescence generation, it is currently constrained by the cost and performance of components; at the same time, the excitation efficiency of longer wavelength is low, and the possibility of burning the sample is also increased.
目前，便携式拉曼光谱仪消除荧光的主要手段是通过基线拟合进行扣减；虽然在直观上将被荧光抬高的光谱基线扣减至初始位置，但是这种扣减无法在机理上或实质上做出解释，难以保证数据处理的真实性和合理性。而其他符合机理的荧光扣减，对于设备硬件都提出了要求，例如，双波长激发，利用拉曼光的迁移性和荧光的相对稳定，消除荧光，但都存在增加设备造价和复杂性的缺点。 At present, the main method for portable Raman spectrometers to eliminate fluorescence is subtraction through baseline fitting; although it is intuitive to subtract the spectral baseline raised by fluorescence to the original position, this subtraction cannot be mechanistically or substantially It is difficult to guarantee the authenticity and rationality of data processing. Other fluorescence subtractions that conform to the mechanism have put forward requirements for equipment hardware, such as dual-wavelength excitation, using the mobility of Raman light and the relative stability of fluorescence to eliminate fluorescence, but all have the disadvantages of increasing equipment cost and complexity. .
发明内容 Contents of the invention
本发明要解决的技术问题是：提供一种能保证数据处理的真实性和合理性、且设备简单、造价低的利用荧光褪色效应消除拉曼光谱中基线干扰的方法。 The technical problem to be solved by the present invention is to provide a method for eliminating baseline interference in Raman spectrum by using fluorescence fading effect, which can ensure the authenticity and rationality of data processing, and has simple equipment and low cost.
解决上述技术问题的技术方案是：一种利用荧光褪色效应消除拉曼光谱中基线干扰的方法，该方法是利用不同激发时刻的光谱差异，找出荧光的度量值，从原始的混合了荧光和拉曼光的光谱中，扣除该度量值，得到纯净的拉曼光谱信号。 The technical solution to solve the above technical problems is: a method for eliminating baseline interference in Raman spectra by using the fluorescence fading effect. This method is to use the spectral difference at different excitation moments to find out the measurement value of the fluorescence, from the original mixture of fluorescence and In the spectrum of Raman light, the measured value is subtracted to obtain a pure Raman spectrum signal.
本发明的进一步技术方案是：该方法的主要内容包括：通过测量样品的光谱信号S，计算系列光谱信号的差值ΔS及其平均值ΔS’，并对差值平均值ΔS’作降噪处理，经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs；从样品光谱信号中逐步扣减荧光的度量值Fs，判断扣减是否达到要求，如果是，则消除了拉曼光谱中基线干扰，得到纯净的拉曼光谱信号。 The further technical solution of the present invention is: the main content of the method includes: by measuring the spectral signal S of the sample, calculating the difference ΔS and its average value ΔS' of the series of spectral signals, and performing noise reduction processing on the difference average value ΔS' , the difference average ΔS' after noise reduction is used as the measurement value Fs of the fluorescence contained in the sample spectral signal; the measurement value Fs of fluorescence is gradually deducted from the sample spectral signal to determine whether the deduction meets the requirements, and if so, eliminate The baseline interference in the Raman spectrum is eliminated, and a pure Raman spectrum signal is obtained.
本发明的再进一步技术方案是：所述的判断扣减是否达到要求是通过选择拉曼迁移2000cm-1或2500cm-1周围50-100个数据点的平坦性来判断扣减是否达到要求。 A further technical solution of the present invention is: the determination of whether the deduction meets the requirement is to judge whether the deduction meets the requirement by selecting the flatness of 50-100 data points around the Raman migration 2000cm −1 or 2500cm −1 .
本发明的再进一步技术方案是：该方法包括如下步骤： A further technical solution of the present invention is: the method comprises the steps of:
①测量样品信号： ① Measure the sample signal:
采用拉曼激光探头，固定焦点、焦距，设定积分时间，连续采取并记录5-10组样品的光谱信号S； Using Raman laser probe, fixed focus and focal length, set integration time, continuously collected and recorded spectral signals S of 5-10 groups of samples;
②计算系列光谱信号的差值： ②Calculate the difference of a series of spectral signals:
按照采集时间的先后，将前一个光谱信号减去后一个光谱信号，得到系列光谱信号的差值ΔS； According to the order of acquisition time, the previous spectral signal is subtracted from the latter spectral signal to obtain the difference ΔS of the series of spectral signals;
③求取ΔS’： ③ Find ΔS':
计算每个差值ΔS间的相关系数，将相关系数大于0.98的差值ΔS选出来并进行平均；得到差值平均值ΔS’； Calculate the correlation coefficient between each difference ΔS, select and average the difference ΔS with a correlation coefficient greater than 0.98; obtain the average value of the difference ΔS';
④对差值平均值ΔS’降噪： ④Denoising the average value of the difference ΔS':
对差值平均值ΔS’作降噪处理，经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs； Perform noise reduction processing on the difference average value ΔS', and the difference value average value ΔS' after noise reduction is used as the measurement value Fs of the fluorescence contained in the sample spectral signal;
⑤从样品光谱信号中逐步扣减荧光的度量值Fs： ⑤ Gradually subtract the fluorescence measurement value Fs from the sample spectral signal:
从样品光谱信号中逐步扣减荧光的度量值Fs，选择拉曼迁移2000cm-1或2500cm-1处作为扣减完全标志，选取该两处的任一处周围50-100个数据点范围内的平坦性判断扣减是否达到要求，如果是，则消除了拉曼光谱中基线干扰，得到纯净的拉曼光谱信号。 Gradually subtract the measurement value Fs of fluorescence from the sample spectral signal, select the Raman migration of 2000cm -1 or 2500cm -1 as the sign of complete subtraction, and select any of the two places within the range of 50-100 data points Flatness judges whether the subtraction meets the requirements, and if so, the baseline interference in the Raman spectrum is eliminated, and a pure Raman spectrum signal is obtained.
本发明的再进一步技术方案是：所述的步骤④对差值平均值ΔS’降噪中，采用滤波器进行降噪，选择2阶滤波，窗口宽度为整体信号长度的1/10。 A further technical solution of the present invention is: in the step 4. in the noise reduction of the difference average value ΔS', a filter is used for noise reduction, and a second-order filter is selected, and the window width is 1/10 of the overall signal length.
本发明的再进一步技术方案是：所述的步骤⑤中，所述的平坦性判断的方法是以拉曼迁移2000cm-1或2500cm-1处周围50-100个数据点范围内的数据点起伏最大值和数据点起伏最小值的差，作为基线平坦判断依据。 A further technical solution of the present invention is: in the step ⑤, the method for judging flatness is based on the fluctuation of data points within the range of 50-100 data points around the Raman migration 2000cm- 1 or 2500cm -1 The difference between the maximum value and the minimum value of the data point fluctuation is used as the basis for judging the flatness of the baseline.
荧光褪色（即漂白），指在光的照射下荧光物质所激发出来的荧光强度随着时间推移逐步减弱乃至消失的现象。而强光激发的拉曼信号，通常并不会随光照时间增加出现明显变化。荧光褪色效应，与光照时间相关，时间越长，荧光强度降低越多，但逐渐趋于稳定，多数情况下，不会完全消失。以往作为消除荧光的一种辅助手段，其缺点是需要长达小时级的光褪色时间，除此之外，样品暴露时间过长，会受到环境、光照的影响发生改变，最终反映到测量结果当中。 Fluorescence fading (that is, bleaching) refers to the phenomenon that the fluorescence intensity excited by fluorescent substances gradually weakens or even disappears over time under the irradiation of light. However, the Raman signal excited by strong light usually does not change significantly with the increase of illumination time. The fluorescence fading effect is related to the illumination time. The longer the time is, the more the fluorescence intensity will decrease, but it will gradually stabilize, and in most cases, it will not disappear completely. In the past, as an auxiliary means to eliminate fluorescence, its disadvantage is that it takes hours for light fading time. In addition, if the sample is exposed for too long, it will be affected by the environment and light and will be changed, which will eventually be reflected in the measurement results. .
本发明的原理为： Principle of the present invention is:
光谱仪直接测量的光谱信号S可定义为拉曼信号R和荧光信号F， The spectral signal S directly measured by the spectrometer can be defined as a Raman signal R and a fluorescent signal F,
S=R+F， S=R+F,
实验证实，随光照时间延长，荧光子逐渐减少，即，各点的荧光强度随时间以初始荧光强度F0等比下降， Experiments have confirmed that as the illumination time prolongs, the fluorophores gradually decrease, that is, the fluorescence intensity of each point decreases proportionally with the initial fluorescence intensity F 0 over time,
F=A(t)*F0，其中A(t)为随时间变化的值。 F=A(t)*F 0 , where A(t) is a value varying with time.
某时刻i与后续时刻i+n测量的光谱信号差值ΔS，在相同激光强度下，拉曼光强不变，所以： The spectral signal difference ΔS measured at a certain time i and the subsequent time i+n, under the same laser intensity, the Raman light intensity remains unchanged, so:
ΔS=S(i)-S(i+n)=[A(i)–A(i+n)]*F0，(n=1、2、3…)。 ΔS=S(i)-S(i+n)=[A(i)-A(i+n)]*F 0 , (n=1, 2, 3...).
ΔS与F0成比例关系，也就是说，从光谱信号S中逐步扣减差值ΔS，根据判定原则，就可以消除荧光的影响，得到拉曼信号R。 ΔS is proportional to F0, that is to say, the difference ΔS is gradually deducted from the spectral signal S, and according to the judgment principle, the influence of fluorescence can be eliminated to obtain the Raman signal R.
由于采用上述结构，本发明之利用荧光褪色效应消除拉曼光谱中基线干扰的方法与现有技术相比，具有以下有益效果： Due to the adoption of the above structure, the method of the present invention utilizing the fluorescence fading effect to eliminate baseline interference in the Raman spectrum has the following beneficial effects compared with the prior art:
1.能保证数据处理的真实性和合理性： 1. Can guarantee the authenticity and rationality of data processing:
本发明利用了荧光和拉曼光在强激发光下的不同响应特性，即：在强激发光照射下，荧光强度会随时间下降，而拉曼光强度保持相对稳定。本发明提出，利用不同激发时刻的光谱差异，找出荧光的度量值，从原始的混合了荧光和拉曼光的光谱中，扣除该度量值，得到纯净的拉曼光谱信号。因此，本发明采用强光持续照射导致荧光发射强度下降的褪色效应，结合化学计量学计算，从受到荧光干扰的激发光谱中消除荧光，从而获得更真实的拉曼光谱信号。与既有方法相比，本发明从原理上解决拉曼光谱中荧光干扰消除这一关键问题，提供了一套更合理可靠、更准确高效的解决方法，能保证数据处理的真实性和合理性。 The invention utilizes the different response characteristics of fluorescence and Raman light under strong excitation light, that is, under the irradiation of strong excitation light, the fluorescence intensity will decrease with time, while the Raman light intensity remains relatively stable. The present invention proposes to use the spectrum difference at different excitation times to find out the measurement value of fluorescence, and subtract the measurement value from the original spectrum mixed with fluorescence and Raman light to obtain a pure Raman spectrum signal. Therefore, the present invention uses the fading effect that the intensity of fluorescence emission decreases due to continuous irradiation of strong light, combined with stoichiometric calculations, to eliminate fluorescence from the excitation spectrum interfered by fluorescence, thereby obtaining a more realistic Raman spectrum signal. Compared with the existing methods, the present invention solves the key problem of fluorescence interference elimination in Raman spectroscopy in principle, provides a set of more reasonable, reliable, more accurate and efficient solutions, and can ensure the authenticity and rationality of data processing .
2.所用的设备简单、造价低： 2. The equipment used is simple and low in cost:
由于本发明提出了采用强光持续照射导致荧光发射强度下降的褪色效应，结合化学计量学计算，从受到荧光干扰的激发光谱中消除荧光，从而获得更真实的拉曼光谱信号。本发明无需提高设备的复杂程度，节约仪器造价，未增加测试和仪器成本，符合便携式拉曼光谱仪高效、便捷的仪器特性。 Since the present invention proposes the fading effect of using strong light continuous irradiation to reduce the intensity of fluorescence emission, combined with stoichiometric calculations, fluorescence is eliminated from the excitation spectrum interfered by fluorescence, thereby obtaining a more realistic Raman spectrum signal. The invention does not need to increase the complexity of the equipment, saves the cost of the instrument, does not increase the cost of the test and the instrument, and conforms to the high-efficiency and convenient instrument characteristics of the portable Raman spectrometer.
3.方法简单，易于普及。 3. The method is simple and easy to popularize.
下面，结合附图和实施例对本发明之利用荧光褪色效应消除拉曼光谱中基线干扰的方法的技术特征作进一步的说明。 In the following, the technical features of the method for eliminating baseline interference in Raman spectra by utilizing the fluorescence fading effect of the present invention will be further described in conjunction with the accompanying drawings and examples.
附图说明 Description of drawings
图1：步骤①中测量的样品光谱信号S的示意图， Figure 1: Schematic diagram of the sample spectral signal S measured in step ①,
图2：步骤②中系列光谱信号的差值ΔS的示意图， Figure 2: Schematic diagram of the difference ΔS of the series of spectral signals in step ②,
图3：步骤③中求取差值平均值ΔS’的示意图， Figure 3: Schematic diagram of calculating the average value of the difference ΔS’ in step ③,
图4：步骤④中降噪后的差值平均值ΔS’的示意图， Figure 4: Schematic diagram of the difference average value ΔS’ after noise reduction in step ④,
图5：步骤⑤中从样品光谱信号中扣减荧光的度量值Fs后的示意图。 Fig. 5: Schematic diagram after deducting the fluorescence measurement value Fs from the sample spectral signal in step ⑤.
具体实施方式 detailed description
一种利用荧光褪色效应消除拉曼光谱中基线干扰的方法，该方法是利用了荧光和拉曼光在强激发光下的不同响应特性，即：在强激发光照射下，荧光强度会随时间下降，而拉曼光强度保持相对稳定。本发明提出，利用不同激发时刻的光谱差异，找出荧光的度量值，从原始的混合了荧光和拉曼光的光谱中，扣除该度量值，得到纯净的拉曼光谱信号。该方法的主要内容包括：通过测量样品的光谱信号S，计算系列光谱信号的差值ΔS及其平均值，并对差值平均值ΔS’作降噪处理，经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs；从样品光谱信号中逐步扣减荧光的度量值Fs，选择拉曼迁移2000cm-1或2500cm-1周围50-100个数据点的平坦性判断扣减是否达到要求，如果是，则消除了拉曼光谱中基线干扰，得到纯净的拉曼光谱信号。该方法包括如下步骤： A method for eliminating baseline interference in Raman spectra by using the fluorescence fading effect, which utilizes the different response characteristics of fluorescence and Raman light under strong excitation light, that is, under the irradiation of strong excitation light, the fluorescence intensity will change with time decreased, while the Raman light intensity remained relatively constant. The present invention proposes to use the spectrum difference at different excitation times to find out the measurement value of fluorescence, and subtract the measurement value from the original spectrum mixed with fluorescence and Raman light to obtain a pure Raman spectrum signal. The main content of this method includes: by measuring the spectral signal S of the sample, calculating the difference ΔS and its average value of the series of spectral signals, and performing noise reduction processing on the average value ΔS' of the difference, and the average value of the difference after noise reduction ΔS' is used as the measurement value Fs of the fluorescence contained in the sample spectral signal; the measurement value Fs of the fluorescence is gradually deducted from the sample spectral signal, and the flatness judgment of 50-100 data points around the Raman migration 2000cm -1 or 2500cm -1 is selected Whether the deduction meets the requirements, if so, the baseline interference in the Raman spectrum is eliminated, and a pure Raman spectrum signal is obtained. The method comprises the steps of:
①测量样品信号： ① Measure the sample signal:
采用拉曼激光探头，固定焦点、焦距，设定积分时间，连续采取并记录5-10组样品的光谱信号S； Using Raman laser probe, fixed focus and focal length, set integration time, continuously collected and recorded spectral signals S of 5-10 groups of samples;
②计算系列光谱信号的差值： ②Calculate the difference of a series of spectral signals:
按照采集时间的先后，将前一个光谱信号减去后一个光谱信号，得到系列的差值ΔS；③求取差值平均值ΔS’： According to the order of acquisition time, the previous spectral signal is subtracted from the latter spectral signal to obtain a series of difference ΔS; ③ Find the average value of the difference ΔS’:
计算每个差值ΔS间的相关系数，将相关系数大于0.98的差值ΔS选出来并进行平均；得到差值平均值ΔS’； Calculate the correlation coefficient between each difference ΔS, select and average the difference ΔS with a correlation coefficient greater than 0.98; obtain the average value of the difference ΔS';
④对差值平均值ΔS’降噪： ④Denoising the average value of the difference ΔS':
对差值平均值ΔS’作降噪处理，采用滤波器进行降噪，选择2阶滤波，窗口宽度为整体信号长度的1/10；经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs； Perform noise reduction processing on the difference average value ΔS', use a filter for noise reduction, select a second-order filter, and the window width is 1/10 of the overall signal length; the difference value average value ΔS' after noise reduction is used as the sample spectral signal A measure of the fluorescence involved, Fs;
⑤从样品光谱信号中逐步扣减荧光的度量值Fs： ⑤ Gradually subtract the fluorescence measurement value Fs from the sample spectral signal:
从样品光谱信号中逐步扣减荧光的度量值Fs，选择拉曼迁移2000cm-1或2500cm-1处作为扣减完全标志，选取该两处的任一处周围50-100个数据点范围内的平坦性判断扣减是否达到要求。随着扣减步长增加，该范围内的数据点起伏逐渐下降，如果荧光被过量扣除，起伏又将逐渐增加；因此，以该范围内的数据点起伏最大值和数据点起伏最小值的差，作为基线平坦判断依据。如果已达到要求，则消除了拉曼光谱中基线干扰，得到纯净的拉曼光谱信号。 Gradually subtract the measurement value Fs of fluorescence from the sample spectral signal, select the Raman migration of 2000cm -1 or 2500cm -1 as the sign of complete subtraction, and select any of the two places within the range of 50-100 data points Flatness judges whether the deduction meets the requirements. As the subtraction step increases, the fluctuation of data points in this range gradually decreases, and if the fluorescence is excessively subtracted, the fluctuation will gradually increase again; therefore, the difference between the maximum value of data point fluctuation and the minimum value of data point fluctuation in this range , as the basis for judging the flatness of the baseline. If the requirements have been met, the baseline interference in the Raman spectrum is eliminated, and a pure Raman spectrum signal is obtained.
以下是本发明的具体实施案例： The following are specific implementation cases of the present invention:
实施例一：Embodiment one:
一种利用荧光褪色效应消除蔗糖拉曼光谱中基线干扰的方法，包括以下步骤： A method utilizing fluorescence fading effect to eliminate baseline interference in sucrose Raman spectrum, comprising the following steps:
①测量样品信号： ① Measure the sample signal:
采用拉曼激光探头，采用785nm激光作为激发，测量蔗糖晶体的拉曼光谱，设定积分时间为10s，连续测量10次，并记录所得到的光谱信号S。从图1中可以看出，光谱信号中存在荧光褪色现象，信号强度呈现依次下降； Using a Raman laser probe, using a 785nm laser as excitation, measure the Raman spectrum of sucrose crystals, set the integration time to 10s, measure 10 times continuously, and record the obtained spectral signal S. It can be seen from Figure 1 that there is a phenomenon of fluorescence fading in the spectral signal, and the signal intensity decreases sequentially;
②计算系列光谱信号的差值： ②Calculate the difference of a series of spectral signals:
按照采集时间的先后，将前一个光谱信号减去后一个光谱信号，得到系列的差值ΔS，图2中只列出了其中几个； According to the order of acquisition time, the previous spectral signal is subtracted from the latter spectral signal to obtain a series of difference values ΔS, and only a few of them are listed in Figure 2;
③求取差值平均值ΔS’： ③ Calculate the average value of the difference ΔS':
计算每个差值ΔS间的相关系数，从差值ΔS系列中将不一致（相关系数<0.98）的挑选掉，将相关系数大于或等于0.98的差值ΔS选出来并进行平均，以消除一部分噪声；得到差值平均值ΔS’，作为新的差值。 Calculate the correlation coefficient between each difference ΔS, select the inconsistent (correlation coefficient <0.98) from the difference ΔS series, and select and average the difference ΔS with a correlation coefficient greater than or equal to 0.98 to eliminate part of the noise ; Obtain the difference average ΔS' as the new difference.
④差值平均值ΔS’降噪： ④ Difference average value ΔS’ noise reduction:
对差值平均值ΔS’作降噪处理，采用Savitzky-Golay滤波器进行降噪，选择2阶滤波，窗口宽度为整体信号长度的1/10；经过降噪后的差值平均值ΔS’作为样品光谱信号所包含荧光的度量值Fs； The average value of the difference ΔS' is denoised, and the Savitzky-Golay filter is used for noise reduction. The second-order filter is selected, and the window width is 1/10 of the overall signal length; the average value of the difference ΔS' after denoising is used as The measurement value Fs of the fluorescence contained in the sample spectral signal;
⑤从样品光谱信号中逐步扣减荧光的度量值Fs： ⑤ Gradually subtract the fluorescence measurement value Fs from the sample spectral signal:
从样品光谱信号中逐步扣减荧光的度量值Fs，选择拉曼迁移2000cm-1处作为扣减完全标志，选取该两处的任一处周围100个数据点范围内的平坦性判断扣减是否达到要求，如果是，则消除了拉曼光谱中基线干扰；如果否，则重复步骤⑤。其中基线平坦判断依据是该范围内的数据点起伏最大值和数据点起伏最小值的差。 Gradually subtract the measurement value Fs of fluorescence from the sample spectral signal, select the Raman migration 2000cm -1 as the complete sign of the subtraction, and select the flatness within 100 data points around any of the two places to judge whether the subtraction is Meet the requirements, if yes, eliminate the baseline interference in the Raman spectrum; if not, repeat step ⑤. The baseline flatness is judged based on the difference between the maximum value of the data point fluctuation and the minimum value of the data point fluctuation within the range.
图5中R0为原始测量的光谱图，R1为去荧光后的光谱图，从该图5中可看出，原始信号中所含的荧光已经得到了很好的消除，而拉曼位移和峰强度特征表达更为清晰、合理。本发明在在不增加设备造价和测量附件的情况下，可以快速得到符合实际机理的拉曼真实信号，并消除了荧光所造成的干扰。 In Figure 5, R0 is the original measured spectrum, and R1 is the spectrum after defluorescence. It can be seen from Figure 5 that the fluorescence contained in the original signal has been well eliminated, while the Raman shift and peak The intensity feature expression is clearer and more reasonable. The invention can quickly obtain the real Raman signal conforming to the actual mechanism without increasing equipment cost and measuring accessories, and eliminates the interference caused by fluorescence.
作为本实施例一的一种变换，所述的步骤⑤中也可以是选择拉曼迁移2000cm-1或2500cm-1处作为扣减完全标志，选取该两处的任一处周围50-100个数据点范围内的平坦性判断扣减是否达到要求。 As a transformation of the first embodiment, in the step ⑤, it is also possible to select the Raman migration of 2000cm- 1 or 2500cm -1 as the mark of complete deduction, and select 50-100 around any of the two places The flatness within the range of data points judges whether the deduction meets the requirements.
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