Method for calibrating four-quadrant detector in real time
Technical Field
The invention relates to a calibration method of optical equipment, in particular to a method for calibrating a four-quadrant detector in real time
Background
A four-quadrant photodiode detector (QD) is a photodetector device in which four photodiodes of identical performance are arranged according to a rectangular coordinate requirement, and is commonly used for laser guidance or laser collimation. The method has the characteristics of high sensitivity, high response speed (up to the order of sub-millisecond), high measurement precision, digital storage of acquired signals, easiness in subsequent processing and the like, so that the method is valued and applied by people in the displacement measurement field. The basic structure of a four-quadrant photodiode detector is shown in fig. 1, and it is composed of four photodiodes with the same parameters, which are respectively located in four quadrants of a circular surface. When light shines on the detector, each diode will output a current signal proportional to its surface light power due to photovoltaic effect, I1、I2、I3And I4. When the detected object is microscopicSmall lateral or longitudinal movements, as indicated by the grey circles in fig. 1, will cause a change in the difference in the optical power projected onto the left and right parts of the four-quadrant photodiode detector or the difference in the optical power of the upper and lower parts, and thus a corresponding change in the photocurrent. Difference value delta I of optical power of left and right parts and upper and lower partsxAnd Δ IyCan be expressed as follows:
ΔIx(I1+I4)-(I2+I3) (1)
ΔIy=(I1+I2)-(I3+I4) (2)
delta I in the above formulaxAnd Δ IyRespectively proportional to the amount of lateral movement and the amount of longitudinal movement of the image of the detected object. The photocurrent caused by the object image is converted into voltage through a four-way current-voltage conversion circuit, and then the voltage is calculated through a summing circuit and a difference circuit, and finally an output voltage signal representing the position (x, y) of the object on a plane rectangular coordinate can be obtained.
To accurately measure the displacement of the object, the QD must be calibrated to obtain the conversion relationship between the displacement and the output voltage signal, and the calibration of the conversion relationship between the displacement and the output voltage signal is directly related to the accuracy of the displacement measurement of the object. A conventional calibration method is disclosed in reference 1 "HL Guo, CX Liu, ZL Li, JFDuan, XH Han, BY Cheng and DZ Zhang, Displacement and force measurement sensitivity calibration on optical detectors, in chinese, phys, lett, vol.20, No.6, pp 950-. Fig. 2 shows a variation curve of the output voltage when the four-quadrant photodiode detector and the small ball captured by the optical trap move relatively in the above conventional calibration method.
The conventional calibration method requires a screw adjusting knob for manually adjusting the QD to change the displacement of the QD point by point, thereby generating the movement of an image relative to the QD. The method is inconvenient to operate and long in time consumption, and measurement errors of the final conversion coefficient can be caused by instability of the system, inaccurate QD displacement adjustment and the like, so that the experimental result is further influenced. In addition, the displacement-to-voltage conversion coefficients obtained in the conventional calibration method can only be applied to the same objects as in the QD calibration. If the objects are irregular in shape or different in size, different transformation factors are used. However, in the experimental process, the size and the shape of each real object have certain differences, the conversion coefficient corresponding to each real object in the experiment obtained by using the traditional calibration method has no operability, and the coefficient obtained by adopting the previous calibration inevitably brings errors to the experiment. Finally, the traditional calibration method is not suitable for repeatedly calibrating objects which are easy to damage, such as biological cells and the like, and the application range of the method is limited.
Disclosure of Invention
The invention aims to overcome the defects that the traditional calibration method for the four-quadrant photodiode detector is inconvenient to operate, long in time consumption and easy to cause measurement errors, and thus the four-quadrant photodiode detector is simple to operate and high in measurement precision.
In order to achieve the above object, the present invention provides an optical measurement system, which includes a four-quadrant photodiode detector, and a detected object image generating device; the stepping motor is provided with a glass slide and can drive the glass slide to vibrate; wherein,
the stepping motor is positioned between the four-quadrant photodiode detector and the detected object image generating device, and light rays emitted to the four-quadrant photodiode detector by the detected object image generating device just penetrate through a glass slide glass on the stepping motor.
In the technical scheme, the stepping motor drives the glass slide to vibrate in a square wave mode.
In the technical scheme, the square wave vibration angle of the glass slide driven by the stepping motor is +/-7.5 degrees.
In the above technical solution, the vibration frequency of the stepping motor is 2 Hz.
The invention also provides a method for calibrating the four-quadrant photodiode detector on the optical measurement system in real time, which comprises the following steps:
step 1), starting the stepping motor, and driving the glass slide to perform square wave vibration by the stepping motor;
step 2), the light with the detected object image emitted by the detected object image generating device passes through the glass slide and irradiates on the four-quadrant photodiode detector, and when the glass slide is subjected to square wave vibration, the detected object image moves relative to the four-quadrant photodiode detector to obtain an output voltage signal of the four-quadrant photodiode detector;
and 3) calculating a conversion coefficient between the displacement of the four-quadrant photodiode detector and the output voltage according to the output voltage signal obtained in the step 2) and the displacement of the image of the detected object relative to the four-quadrant photodiode detector when the glass slide is subjected to square wave vibration, so as to realize the calibration of the four-quadrant photodiode detector.
In the above technical solution, in the step 3), the displacement of the detected object image relative to the four-quadrant photodiode detector is obtained by calibration when the glass slide performs square wave vibration, and the calibration includes:
step 3-1), collecting detected object images in a series of optical traps obtained when the glass slide vibrates by using a video charge-coupled device;
and 3-2) obtaining the position of the detected object image by adopting a gray scale gravity center method, thereby obtaining the displacement of the detected object image relative to the four-quadrant photodiode detector.
The invention has the advantages that:
1. the invention directly calibrates the conversion coefficient between the output voltage of the four-quadrant photodiode and the image displacement of the detected object through the relation between the translation light distance of the glass slide and the output signal of the four-quadrant photodiode detector, and does not need to adjust the original optical path system, thereby having the advantages of simple and rapid operation and real-time and accurate acquisition of experimental parameters.
2. The invention can calibrate a specific object in real time, thereby overcoming the defect of one-time calibration and repeated use of the traditional method caused by the problem of the size consistency of the detected object.
3. The invention does not carry out operations such as movement and the like on the detected object, does not influence the detected object and has wide application range.
Drawings
FIG. 1 is a schematic diagram of a four-quadrant photodiode detector;
FIG. 2 is a graph showing the variation of output voltage when a small ball captured by a four-quadrant photodiode detector and an optical trap moves relative to each other, which is obtained by a conventional calibration method;
FIG. 3 is a schematic diagram of the optical path structure of an optical tweezers system employing the method of the present invention in one embodiment;
FIG. 4 is a graph of voltage signals output by the four-quadrant photodiode detector when the glass sheet is vibrated with a square wave.
Description of the drawings
1 halogen lamp 2 first lens 3 first reflector
4 first filter 5 condenser 6 object plane
7 mercury lamp 8 second filter 9 first dichroic mirror
10 objective 11 auxiliary objective 12 second dichroic mirror
13 second lens, 14 ocular and 15 fourth dichroic mirror
16CCD 17 bottom outlet 18 laser
19 first expander lens 20 second expander lens 21 second reflector
22 third expander lens 23 third dichroic mirror 24 fourth expander lens
25 four-quadrant photodiode detector 26 glass slide
Detailed Description
The invention is described below with reference to the accompanying drawings and the detailed description.
In one embodiment of the present invention, a conventional optical tweezers system is taken as an example to illustrate how the method of the present invention can be used in the system.
In fig. 3, a schematic diagram of an optical tweezers system is shown, and it can be seen that the optical tweezers system comprises an inverted research type optical microscope, a Charge Coupled Device (CCD) and a four-quadrant photodiode detector (QD), and further comprises other necessary optical elements, such as lenses, mirrors, etc. Wherein, the inverted research type optical microscope is represented by a dotted frame part in the figure, and comprises a Koehler illumination system which is composed of a Halogen Lamp (Halogen Lamp)1, a first lens 2, a first reflector 3, a first filter 4 and a condenser 5, so as to form uniform bright field illumination on an object plane 6; the mercury lamp 7, the second filter 8 and the first dichroic mirror 9 form a fluorescent lighting system; an objective lens 10, an auxiliary objective lens 11, a second dichroic mirror 12, a second lens 13 and an ocular lens 14 form an imaging system with an infinite optical cylinder length, so that a sample on an object plane 6 forms an amplified virtual image at the ocular lens 14, and meanwhile, a fourth dichroic mirror 15 images the sample on a CCD 16; the bottom exit 17 is used for the introduction of the laser and for the exit of the additional imaging beam path. The 1064nm laser 18 is expanded into parallel light by a first beam expanding lens 19 and a second beam expanding lens 20, and is reflected into an inverted microscope by a second reflector 21, and a third beam expanding lens 22 is coupled with a Tube lens of the microscope, so that the laser is changed into the parallel light. The expanded beam must fill the back pupil of the objective lens to ensure that a sufficient light intensity gradient is generated to form a stable light trap. The objective lens 10, the auxiliary objective lens 11, the third dichroic mirror 23, and the fourth beam expanding lens 24 constitute an imaging system, which images the object plane 6 onto a photosensitive surface of the four-quadrant photodiode detector 25 for dynamic monitoring and recording of the experimental process. In this embodiment, the inverted research optical microscope may employ Leica DMIRB, the CCD may employ CoolSNAP-fx, and the four-quadrant photodiode detector may employ HAMAMATSU 1557-03.
After obtaining the optical tweezers system configured as above, polystyrene beads with a diameter of 1 micrometer can be measured. During this measurement, QDs were used to monitor the displacement information of the polystyrene pellet from the center of the optical trap. As described in the background, before the displacement of polystyrene beads is monitored by QD, calibration of QD is required to obtain the conversion relationship between displacement and output voltage signal. In the method of the present invention, a conventional glass slide 26 having a thickness of 1.0mm is mounted on a stepper motor, and then the glass slide 26 is placed in front of the QDs 25 so that light can pass vertically through and to the QDs 25 when the glass slide 26 is at rest. The stepper motor is adapted to control the glass slide 26 to vibrate in a square wave, so as to ensure that the signal generated by the QD during the square wave vibration is within the linear region (indicated by the thick line segment in fig. 2) of fig. 2, and therefore, in this embodiment, the square wave vibration angle of the glass slide is within ± 7.5 °, and the vibration frequency of the stepper motor is 2 Hz. However, it should be understood by those skilled in the art that the square wave vibration angle of the glass slide, the vibration frequency of the stepping motor, and the thickness of the glass slide are not limited to the above mentioned data, and the skilled person can adjust the vibration angle, the vibration frequency, and the thickness of the glass slide according to the actual requirement.
And a glass slide is placed in a light path in front of the QD, and when the glass slide is driven by a stepping motor to perform square wave vibration of +/-7.5 degrees, according to the principle that light rays are translated after passing through plate glass, emergent light rays move when the glass slide deflects to two different angles, so that images of the polystyrene spheres to be detected are located at two different positions of the QD. According to the operating principle of the QD itself, as shown in fig. 4, the QD generates a series of square wave output voltage signals with a frequency of 2Hz, and the peak-to-peak values of the square wave signals can be obtained according to the output voltage signals. By this operation, data relating to the output voltage is known, and it is obvious that data relating to the displacement is also required to obtain the conversion coefficient of the displacement of the QD with the output voltage.
When the distance of the small ball image moving relative to the QD is calibrated when the glass slide is vibrated by square waves, a video CCD can be used for replacing the QD, then polystyrene small ball images in a series of optical traps obtained when the glass slide is vibrated are collected, and the positions of the polystyrene small ball images can be obtained by processing the polystyrene small ball images by a gray scale gravity center method. The conditions such as the size of the small ball are changed for measuring for a plurality of times, the repeatability of the system is found to be good, and the image position is irrelevant to the factors such as the size of the small ball. And (3) statistically averaging the multiple measurement data to obtain: the polystyrene bead image was moved relative to the QDs by a distance of 91.6 + -0.9 μm. The information of the displacement of the polystyrene spheres from the center of the optical trap is obtained through the operation. In the present embodiment, the measurement of the data related to the displacement is implemented by the above method, but it should be understood by those skilled in the art that the measurement of the displacement is not limited to the above method, and other methods in the prior art can be used as well. In addition, it should be noted that, since the repeatability of the stepping motor is very good, as long as the stepping motor swings at the same angle every time, the displacement is a fixed value, so that the calibration of the moving distance of the bead image relative to the QD when the glass slide is subjected to square wave vibration only needs to be performed once, and is not required to be repeated.
After the peak value of the square wave signal and the moving distance of the polystyrene bead image relative to the QD are obtained, the conversion coefficient of the displacement of the QD and the output voltage can be rapidly obtained. As shown in fig. 4, the peak-to-peak value of the square wave signal obtained in this example is about 250mV, and the moving distance of the polystyrene bead image relative to the QD is 91.6 ± 0.9 μm, so that the displacement-voltage conversion coefficient of the polystyrene bead image relative to the QD is calculated to be 5.47 ± 0.05mV/μm. Whereas, if a calibration curve as shown in FIG. 2 is obtained by a conventional method of adjusting the position of QDs, and a linear portion thereof is fitted, the shift-to-voltage conversion coefficient of the image of polystyrene beads with respect to QDs is 5.46 mV/. mu.m. The difference of the results obtained by the two methods is only 0.01 mV/mum and less than 0.2 percent, which proves that the method is accurate and reliable, the relative error is less than 1 percent, the method can be used for rapidly obtaining accurate experimental parameters in real time, the experimental precision is improved, and the simplicity and convenience of operation are greatly improved.
In the above embodiments, the implementation and effect of the method of the present invention are described by taking a common optical tweezers system as an example, and it should be understood by those skilled in the art that the method of the present invention is not limited to be used in the optical tweezers system, and the method of the present invention can also be used in other measurement systems, such as a measurement system for cantilever movement of an atomic force microscope, a precision measurement system for workpiece movement in industrial machining, and the like.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and are not limited. Although the present invention has been described in detail with reference to the embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims.