JP5378187B2 - Scanning microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide the technology of forming an image in which image distortion and displacement are suppressed, without excessively complicating the structure and control of a device. <P>SOLUTION: A scanning type microscope 1 includes a sampling clock generating circuit 18 for generating a sampling clock signal that indicates the timing of sampling an image detection signal from a scanning synchronous signal that has a period equal to the period of the sinusoidal oscillation of a two-dimensional scanning section 12. Then, the sampling clock generating circuit 18 includes a temperature correction signal generating section 19 for generating a temperature correction signal that corrects a sampling clock signal according to a set temperature. Accordingly, without excessively complicating the structure and control of the device, the scanning type microscope 1 forms an image in which image distortion and displacement are suppressed. <P>COPYRIGHT: (C)2011,JPO&amp;INPIT

Description

  The present invention relates to a technique of a scanning microscope, and more particularly to a technique for sampling an image signal of a scanning microscope.

  2. Description of the Related Art Conventionally, there is known a scanning microscope that irradiates a sample with light collected in a spot shape and detects the transmitted light, reflected light, or fluorescence generated from the sample with a photodetector. Examples of such a scanning microscope include a confocal laser microscope using a confocal stop and a multiphoton excitation microscope for detecting fluorescence generated through a multiphoton process.

  In a scanning microscope in which light is focused on a sample in a spot shape, in order to generate an image of the sample, a scanning unit that scans the sample in two dimensions by moving the focusing position in two dimensions on the sample is provided. Necessary. As such scanning means, a galvano mirror or a resonant scanner is usually used in the X direction, and a servo galvano mirror is used in the Y direction.

  When a resonant galvanometer mirror or resonant scanner is used as the scanning means, the scanning speed changes in a sine wave shape, so if the image detection signal from the photodetector is sampled at regular intervals, an image with distortion or deviation is generated. End up. Therefore, the control circuit of the scanning unit generates a synchronization signal (hereinafter referred to as a sampling clock signal) indicating the timing for sampling the image detection signal in accordance with the scanning speed of the scanning unit. The image generation unit generates an image by sampling the image detection signal in synchronization with the input of the sampling clock signal.

  Hereinafter, the condensing position on the sample controlled by the control circuit of the scanning means is referred to as “optical scanning position”. The position of the image obtained by sampling the image detection signal in accordance with the sampling clock signal generated by the control circuit of the scanning unit is referred to as “electrical sampling position”.

  However, in practice, if the control circuit of the scanning means outputs the scanning means driving signal that determines the optical scanning position and the sampling clock signal that determines the electrical sampling position at the same timing, the operation delay of the scanning means and the electric circuit system Due to the time delay, the optical scanning position and the electrical sampling position may be misaligned. In this case, an image with distortion or shift is generated.

  As a technique for suppressing such distortion and shift of an image, the technique disclosed in Patent Document 1 can be used. In the technique disclosed in Patent Document 1, means for detecting the center of the optical scanning range is provided, and the center of the optical scanning range and the center of the electrical sampling range are compared for each scanning of one line. Based on the comparison result, the electrical sampling range is corrected so that the center of the optical scanning range and the center of the electrical sampling range coincide. Thereby, an image in which distortion and shift are suppressed can be generated.

JP 2000-39560 A

  In the technique disclosed in Patent Document 1, both the control for generating the sampling clock and the control for detecting the center of the optical scanning range are performed for each line. For this reason, an error caused by the control for generating the sampling clock and an error caused by the control for detecting the center of the optical scanning range are duplicated at the same timing. In this case, since both errors simultaneously affect the control for correcting the deviation between the optical scanning position and the electrical sampling position, the control becomes complicated.

  Further, since the technique disclosed in Patent Document 1 requires a means for detecting the center of the optical scanning range, the apparatus becomes large and the structure becomes complicated. In addition, the manufacturing cost tends to increase accordingly. Furthermore, since a part of the light to be irradiated onto the sample is used for detecting the center of the optical scanning range, the utilization efficiency of the light emitted from the light source is lowered.

  By the way, the difference between the optical scanning position and the electrical sampling position caused by a delay in the scanning means or the electric circuit system does not depend on the difference in individual characteristics between apparatuses, that is, on the environment where the individual difference of apparatuses is a main factor. Deviations (hereinafter referred to as individual difference deviations) and environmentally dependent deviations (hereinafter referred to as temperature deviations), which are mainly caused by temperature changes, can be broadly classified. Considering that the individual difference deviation based on the temperature assumed in the usage environment (hereinafter referred to as the reference temperature) is corrected before shipment, the main factor of the deviation in the usage environment is the standard. This is considered to depend on the difference between the temperature and the temperature of the actual usage environment (hereinafter referred to as the usage temperature).

  Based on the above situation, the present invention corrects the deviation between the temperature-dependent optical scanning position and the electrical sampling position, thereby preventing image distortion without overcomplicating the structure and control of the apparatus. It is an object of the present invention to provide a technique for generating an image in which deviation is suppressed.

According to a first aspect of the present invention, a scanning means for moving a condensing position of illumination light irradiated on a sample by sine wave vibration, and observation light obtained by irradiating the sample with illumination light are detected. And a sampling clock generating means for generating a sampling clock signal indicating a timing for sampling the image detection signal from a scanning synchronization signal having a period equal to the period of the sine wave vibration. Sampling the image detection signal based on the sampling clock signal, and generating an image of the sample, and the sampling clock generating means generates a temperature correction signal for correcting the sampling clock signal according to the set temperature. a temperature correction means for generating for the temperature correction information storage means for correcting information is stored corresponding to the set temperature The sine wave signal converted from the scanning synchronization signal is inputted, the frequency variable oscillation means for outputting the sampling clock signal, the phase shift means for changing the phase of the sine wave signal, and the amplitude of the sine wave signal. seen including a gain adjustment means for varying the said temperature correction means, from the temperature correction information storing means, are inputted to the correction information corresponding to the set temperature, in response to the set temperature, the gain adjusting means Provided is a scanning microscope for inputting the temperature correction signal for changing the amplitude of the sine wave signal .

According to a second aspect of the present invention, scanning means for moving a condensing position of illumination light applied to a sample by sine wave vibration, and observation light obtained by irradiating the sample with the illumination light are detected, Light detection means for converting the observation light into an image detection signal, which is an electrical signal, and a sampling clock signal indicating timing for sampling the image detection signal from a scanning synchronization signal having a period equal to the period of the sine wave vibration Sampling clock generation means; and image generation means for sampling the image detection signal based on the sampling clock signal and generating an image of the sample, the sampling clock generation means according to a set temperature, Temperature correction means for generating a temperature correction signal for correcting the sampling clock signal, and corresponding to the set temperature Temperature correction information storage means for storing positive information, and frequency variable oscillation means for inputting the sine wave signal converted from the scanning synchronization signal and outputting the sampling clock signal, and the temperature correction means, A scanning microscope that receives the correction information corresponding to the set temperature from the temperature correction information storage unit and changes the amplitude of the sine wave signal input to the variable frequency oscillation unit according to the set temperature. provide.

According to a third aspect of the present invention, in the scanning microscope according to the first aspect, the temperature correction unit causes the phase shift unit to change the phase of the sine wave signal according to the set temperature. A scanning microscope for inputting a temperature correction signal is provided.

According to a fourth aspect of the present invention, in the scanning microscope according to the second aspect, the temperature correction unit changes the phase of the sine wave signal input to the frequency variable oscillation unit according to the set temperature. A scanning microscope to be changed is provided.

According to a fifth aspect of the present invention, in the scanning microscope according to the second aspect, the sampling clock generation means further includes a phase shift means for changing a phase of the sine wave signal, and an amplitude of the sine wave signal. And a gain adjusting means for changing the scanning microscope.

According to a sixth aspect of the present invention, in the scanning microscope according to the fifth aspect, the temperature correction unit causes the phase shift unit to change the phase of the sine wave signal according to the set temperature. A scanning microscope for inputting a temperature correction signal is provided.

According to a seventh aspect of the present invention, in the scanning microscope according to the fifth aspect or the sixth aspect, the temperature correction unit sends the gain adjustment unit with the sine wave signal according to the set temperature. A scanning microscope for inputting the temperature correction signal for changing the amplitude is provided.
An eighth aspect of the present invention, in the scanning microscope according to any one of the first aspect to the seventh aspect, further provides a scanning microscope that includes a temperature sensor.

  According to a ninth aspect of the present invention, there is provided the scanning microscope according to the eighth aspect, wherein the temperature sensor detects the temperature of the sampling clock generating means.

  According to a tenth aspect of the present invention, in the scanning microscope according to the ninth aspect, the temperature correction means generates a temperature correction signal for correcting the sampling clock signal in accordance with the temperature detected by the temperature sensor. Provide a microscope.

  In an eleventh aspect of the present invention, in the scanning microscope according to the ninth aspect, the correction information corresponding to the temperature detected by the temperature sensor is further notified that the temperature correction information storage means is not stored. A scanning microscope including a notification unit is provided.

  A twelfth aspect of the present invention is the scanning microscope according to any one of the ninth to eleventh aspects, further comprising a temperature changing means for changing the temperature of the sampling clock generating means. I will provide a.

  According to a thirteenth aspect of the present invention, in the scanning microscope according to any one of the first to twelfth aspects, the sampling clock generation means includes a sampling clock according to the individual characteristics of the scanning microscope. Provided is a scanning microscope in which an adjustment signal for adjusting a signal is input.

  A fourteenth aspect of the present invention provides the scanning microscope according to any one of the first to thirteenth aspects, further comprising a laser light source that emits a laser beam as illumination light. To do.

  ADVANTAGE OF THE INVENTION According to this invention, the technique which produces | generates the image by which distortion and the shift | offset | difference of an image were suppressed can be provided, without making the structure and control of an apparatus excessively complicated.

1 is a diagram illustrating a configuration of a scanning microscope according to Example 1. FIG. FIG. 3 is a diagram illustrating a configuration of a sampling clock generation circuit according to the first embodiment. FIG. 6 is a diagram illustrating a scanning unit driving signal and a scanning synchronization signal output from the two-dimensional scanning control circuit according to the first embodiment. It is a figure for demonstrating an example of image degradation by the shift | offset | difference of an optical scanning position and an electrical sampling position. It is a flowchart which shows the collection procedure of correction information. It is a figure for demonstrating the method of calculating an interpolation type and the amount of phase shifts. FIG. 6 is a diagram illustrating a configuration of a digital circuit used in a sampling clock generation circuit according to a second embodiment. It is a figure for demonstrating the difference of the characteristic of the phase shift circuit comprised as an analog circuit, and the characteristic of the phase shift circuit comprised as a digital circuit. FIG. 10 is a diagram illustrating a configuration of a sampling clock generation circuit according to a third embodiment. It is a figure for demonstrating an example of image degradation by the shift | offset | difference of an optical scanning position and an electrical sampling position. FIG. 10 is a diagram for explaining a configuration and an operation of an absolute value circuit unit included in a sampling clock generation circuit according to a third embodiment. 6 is a diagram illustrating a configuration of a scanning microscope according to Example 4. FIG.

  Embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is a diagram illustrating the configuration of a scanning microscope according to the present embodiment. A scanning microscope 1 illustrated in FIG. 1 is a confocal laser microscope. In this embodiment, the illumination light is laser light, and the observation light is reflected light from the sample.
First, the configuration of the scanning microscope 1 will be described.

  The scanning microscope 1 irradiates a sample 3 with laser light and outputs an image detection signal corresponding to the intensity of reflected light from the sample 3, a stage 4 on which the sample 3 is arranged, and a laser on the sample 3. An objective lens 5 for condensing light, a revolver 6 to which the objective lens 5 is attached, a computer 7 that samples an image detection signal and generates an image of the sample 3, and a monitor 8 that displays the image of the sample 3 It consists of

  The revolver 6 is provided with a plurality of objective lenses having different magnifications. In FIG. 1, the objective lens 5 is disposed on the optical path. However, by rotating the revolver 6, an objective lens having a desired magnification can be disposed on the optical path.

  The microscope main body 2 has a laser light source 9 that emits laser light, a mirror 10, a half mirror 11, and an optical scanning position that is a condensing position of the laser light irradiated on the sample 3 by deflecting the laser light. A two-dimensional scanning unit 12 that moves in a direction orthogonal to the lens 5, a mirror 13, a lens 14 that collects reflected light, and a focal position of the lens 14 that is optically conjugate with the focal position of the objective lens 5. A confocal stop 15 having a pinhole, a photodetector 16 that converts the reflected light that has passed through the confocal stop 15 into an electrical signal, a two-dimensional scanning control circuit 17 that controls the two-dimensional scanning unit 12, and an optical And a Z-axis scanning unit 20 that moves the scanning position in the optical axis direction of the objective lens 5.

  The two-dimensional scanning control circuit 17 includes a sampling clock generation circuit 18 that is a sampling clock generation means. The sampling clock generation circuit 18 generates a sampling clock signal indicating the timing for sampling the image detection signal, which is an electric signal output from the photodetector 16.

  The sampling clock generation circuit 18 includes a temperature correction signal generation unit 19 that generates a temperature correction signal for correcting the sampling clock signal in accordance with a set temperature that can be arbitrarily set for the scanning microscope 1. . The temperature correction signal generator 19 is a temperature correction unit that corrects a temperature shift depending on the temperature among the shift between the optical scanning position and the electrical sampling position. As the set temperature, the use temperature that is the temperature of the actual use environment is usually used. Thereby, since the difference between the reference temperature and the use temperature can be recognized, the temperature correction signal generator 19 can generate an appropriate signal for correcting the temperature shift.

  In FIG. 1, an example in which the sampling clock generation circuit 18 is included in the two-dimensional scanning control circuit 17 is shown, but the present invention is not limited to this. The sampling clock generation circuit 18 may be provided outside the two-dimensional scanning control circuit 17 as long as it can receive a scanning synchronization signal described later output from the two-dimensional scanning control circuit 17.

  The half mirror 11 is an optical path dividing unit for guiding laser light to the sample 3 and guiding reflected light to the photodetector 16. Instead of the half mirror 11, a polarization beam splitter (PBS) that separates incident light by polarization components may be used.

  The two-dimensional scanning unit 12 is a scanning unit for scanning the sample 3, and for example, a resonant scanner that is an X-direction scanning unit that moves an optical scanning position in the X direction and an optical scanning position that moves in the Y direction. And a servo type galvanometer mirror which is a Y-direction scanning means.

  Although the two-dimensional scanning unit 12 in which the X direction scanning unit and the Y direction scanning unit are combined is illustrated in FIG. 1, the present invention is not limited to this. The X direction scanning unit and the Y direction scanning unit may be arranged separately in the microscope main body 2.

  In the two-dimensional scanning unit 12, any one of the X direction scanning unit and the Y direction scanning unit may be operated by sinusoidal vibration. For this reason, the two-dimensional scanning unit 12 is not limited to the above-described configuration, and may be configured using, for example, a resonant galvanometer mirror instead of the resonant scanner as the X-direction scanning unit. In addition, a servo galvanometer mirror may be used as the X direction scanning unit, and a resonance type galvano mirror or a resonant scanner may be used as the Y direction scanning unit.

As the photodetector 16, for example, a photomultiplier (PMT), a photodiode (Photodiode: PD), an avalanche photodiode (Avalanche Photodiode: APD), or the like can be used.
Next, the operation of the scanning microscope 1 will be described.

  The laser light emitted from the laser light source 9 reflects the mirror 10 and enters the half mirror 11. A part of the laser light incident on the half mirror 11 passes through the half mirror 11 and enters the two-dimensional scanning unit 12. The laser light emitted from the two-dimensional scanning unit 12 is reflected by the mirror 13, collected by the objective lens 5 attached to the revolver 6, and irradiated onto the sample 3.

  Reflected light obtained by irradiating the sample with laser light is incident on the objective lens 5, passes through the mirror 13 and the two-dimensional scanning unit 12, and enters the half mirror 11. Part of the reflected light incident on the half mirror 11 is reflected by the half mirror 11 and collected by the lens 14. Since the focal position of the lens 14 and the focal position of the objective lens 5 have an optically conjugate relationship, unnecessary light generated from other than the optical scanning position on the sample 3 is blocked by the confocal stop 15. The As a result, only the reflected light generated from the optical scanning position is detected by the photodetector 16 which is a light detection means, and an image detection signal is output to the computer 7.

  Thus, since only the reflected light generated from the optical scanning position is detected in the scanning microscope 1, the two-dimensional scanning unit 12 moves the optical scanning position and scans the sample 3 two-dimensionally. Specifically, for example, the X direction scanning unit is reciprocated once in the X direction, and then the Y direction scanning unit is moved in the Y direction. By repeating this, the sample 3 can be scanned two-dimensionally by moving the optical scanning position. The computer 7 serving as an image generation unit samples the image detection signal based on the sampling clock signal, generates an image of the sample 3, and displays the image on the monitor 8.

  In addition, since the scanning microscope 1 uses X-direction scanning means that operates by sinusoidal vibration, the scanning speed changes like a sine wave. For this reason, it is necessary to change the sampling interval of the image detection signal by the computer 7 in accordance with the scanning speed.

  Therefore, the two-dimensional scanning control circuit 17 operates the X-direction scanning unit of the two-dimensional scanning unit 12 by sine wave vibration and outputs a scanning synchronization signal having a period equal to the period of the sine wave vibration to the sampling clock generation circuit 18. To do. Then, the sampling clock generation circuit 18 generates a sampling clock signal corresponding to the scanning speed from the input scanning synchronization signal, so that the optical scanning position and the electrical sampling position coincide with each other, and there is no distortion or misalignment. Can be generated.

  One scanning synchronization signal corresponds to one cycle of sinusoidal vibration of the X-direction scanning unit. Therefore, the sampling clock signal generated from one scanning synchronization signal includes the forward sampling clock signal output in the forward direction of the X-direction scanning means for generating the image of the sample 3 and the X-direction scanning means at the scanning start position. A return sampling clock signal output on the return path of the X-direction scanning means to be returned to is included.

Sampling with one sampling clock signal corresponds to image acquisition for one pixel. For this reason, each of the forward sampling clock signal and the backward sampling clock signal generated from one scanning synchronization signal includes the same number of sampling clock signals as the number of pixels in the X direction of the image. Therefore, for example, if the number of pixels in the X direction is 1024, 2048 sampling clock signals are generated from one scanning synchronization signal.
Next, the configuration and operation of the sampling clock generation circuit 18 will be described in detail.

  FIG. 2 is a diagram illustrating a configuration of the sampling clock generation circuit according to the present embodiment. FIG. 3 is a diagram illustrating scanning means driving signals and scanning synchronization signals output from the two-dimensional scanning control circuit 17 according to the present embodiment.

  The sampling clock generation circuit 18 is a sampling clock generation unit that generates and outputs a sampling clock signal from the scanning synchronization signal output from the two-dimensional scanning control circuit 17.

  Although not described in detail below, the two-dimensional scanning control circuit 17 causes the sampling clock generation circuit 18 to generate a sampling clock signal, generates an image valid signal, and outputs it to the computer 7.

  The image valid signal includes an X-direction image valid signal (hereinafter referred to as an XDE signal) and a Y-direction image valid signal (hereinafter referred to as a YDE signal). For example, the XDE signal is a signal indicating whether the X-direction scanning unit is within or outside the range where the sampling of the image detection signal can be effectively performed. The YDE signal is a signal indicating whether the Y-direction scanning unit is within or outside the range where the sampling of the image detection signal can be effectively performed.

Accordingly, when both the XDE signal and the YDE signal indicate a valid state (within range), the computer 7 samples the image detection signal according to the sampling clock signal and generates an image.
Such an image valid signal may be generated by the two-dimensional scanning control circuit 17 and output to the computer 7.

  2 includes an input signal generation unit 21, a phase shift circuit 22, a position feedback unit 23, an absolute value circuit unit 24, a VCO unit 25, a counter 26, a D / D The A converter 27, the comparator 28, the phase comparison unit 29, the loop filter 30, the temperature correction signal generation unit 19, and the temperature correction information storage unit 31 are configured.

The scanning synchronization signal output from the two-dimensional scanning control circuit 17 is input to the input signal generation unit 21. As illustrated in FIG. 3, the scanning synchronization signal is a square wave signal having a period equal to the period of the scanning unit driving signal which is a speed signal of the X direction scanning unit. The period of the scanning means driving signal is equal to the period of the sine wave vibration of the X direction scanning means. For this reason, the period of the scanning synchronization signal is equal to the period of the sine wave vibration of the X-direction scanning unit. For example, if the period of the sine wave signal of the X-direction scanning unit is 4 kHz, the period of the scanning synchronization signal is also 4 kHz. .
Since the scanning synchronization signal is a square wave signal, it can be regarded as a timing signal synchronized with the timing when the speed of the X-direction scanning unit becomes zero.

  The input signal generation unit 21 converts the scanning synchronization signal, which is a square wave signal, into a sine wave signal having the same period and outputs the sine wave signal to the phase shift circuit 22. The input signal generation unit 21 is configured as an analog circuit including a band pass filter, for example. The bandpass filter converts the square wave signal into a sine wave signal by removing components that contribute to the rising and falling edges of the square wave signal.

  The phase shift circuit 22 is a phase shift means for changing the phase of the sine wave signal generated by the input signal generation unit 21 and changes the phase of the sine wave signal according to the adjustment signal input together with the sine wave signal. The sine wave signal whose phase has been changed by the phase shift circuit 22 is output to the position feedback unit 23. The phase shift circuit 22 is configured as an analog circuit including, for example, an all-pass filter and a potentiometer for changing a resistance value that determines a time constant of the all-pass filter.

  The adjustment signal is a signal set in advance before the scanning microscope 1 is shipped, and is a signal for correcting the individual difference deviation under the reference temperature by changing the phase of the sine wave signal.

The position feedback unit 23 outputs a sine wave signal obtained by taking a difference between a sine wave signal output from the phase shift circuit 22 and an output signal from a D / A converter 27 described later to the absolute value circuit unit 24. To do.
The absolute value circuit unit 24 is a circuit for full-wave rectifying the input signal, and full-wave rectifies the sine wave signal output from the position feedback unit 23 and outputs the sine wave signal to the VCO unit 25.

  The VCO unit 25 is a circuit that performs voltage-frequency conversion, and is a frequency variable oscillation unit that outputs a clock signal having a frequency proportional to the input voltage. The VCO unit 25 outputs a clock signal having a frequency proportional to the voltage of the sine wave signal having a full-wave rectified waveform to the computer 7 as a sampling clock signal.

  The sampling clock signal is also output to the counter 26. The counter 26 counts the sampling clock signal, and the digital signal output from the counter 26 is converted into an analog signal by the D / A converter 27. Since the D / A conversion in the D / A converter 27 is performed in synchronization with the output from the VCO unit 25, the output signal is an analog signal that reproduces the waveform of the analog signal input to the VCO unit 25. Become. More precisely, since the waveform before full wave rectification processing is reproduced, the waveform of the analog signal input to the absolute value circuit unit 24 is reproduced.

  An analog signal that reproduces the input waveform in this way is output to the position feedback unit 23, and the output content from the VCO unit 25 is fed back, so that an error with respect to the nonlinearity of the VCO unit 25 and an instruction (that is, an input signal) It can be corrected.

  The counter 26 generates an XDE signal by latch means (not shown) and outputs it to the computer 7. The YDE signal is generated by a Y-direction drive control circuit (not shown) included in the two-dimensional scanning control circuit 17 and is output to the computer 7 in the same manner as the XDE signal.

  The counter 26 further generates a timing signal (hereinafter referred to as a first timing signal) every line, that is, every time the number of pixels in the X direction is counted, and outputs the timing signal to the phase comparison unit 29.

  The comparator 28 compares the output from the position feedback unit 23 with 0 V, generates a timing signal (hereinafter referred to as a second timing signal) indicating the timing when the output from the position feedback unit 23 becomes 0 V, and compares the phase. To the unit 29.

  The phase comparator 29 compares the phase of the first timing signal with the phase of the second timing signal, and outputs a signal indicating a phase error (hereinafter referred to as a phase error signal) to the loop filter 30. The loop filter 30 outputs a signal that changes the amplitude of the sine wave signal according to the phase error signal to the input signal generation unit 21. Thus, the loop filter 30 changes the gain of the input signal generation unit 21 by changing the amplitude of the sine wave signal generated by the input signal generation unit 21.

  Normally, until the control of the sampling clock generation circuit 18 is stabilized, the number of sampling clock signals output from the VCO unit 25 for one scan synchronization signal input does not match the predetermined number of times required for sampling. That is, for example, when the number of pixels in the X direction is 1024, the number of output sampling clock signals needs to be 2048 in total including the forward and backward sampling clock signals, but is not 2048.

However, by outputting the first timing signal and the second timing signal to the phase comparison unit 29 in this way and feeding back the phase shift between the sine wave signal and the sampling clock signal, the amplitude of the sine wave signal is The timing signal and the second timing signal are stable in phase. The state in which the phases of the first timing signal and the second timing signal coincide with each other is a state in which a number of sampling clock signals equal to the number of pixels in the X direction are generated in synchronization with the half-cycle sine wave signal. For this reason, by such feedback, the number of sampling clock signals output from the VCO unit 25 matches the predetermined number of times while maintaining a coarse / dense state necessary for sampling.
As described above, the optical scanning position and the electrical sampling position can be matched under the reference temperature.

  The sampling clock generation circuit 18 according to the present embodiment includes a temperature correction signal generation unit 19 and a temperature correction information storage unit 31. Then, the temperature correction signal generator 19 generates a temperature correction signal that is a signal for correcting the temperature deviation by changing the phase of the sine wave signal, and outputs the temperature correction signal to the phase shift circuit 22, so that the use temperature is changed. Even when the temperature is different from the reference temperature, the optical scanning position and the electrical sampling position can be matched.

  The temperature correction information storage unit 31 is temperature correction information storage means for storing correction information corresponding to the set temperature. The correction information is, for example, a phase change amount (hereinafter referred to as a phase shift amount), and the temperature correction information storage unit 31 stores a set temperature and a corresponding phase shift amount.

  The set temperature may be stored as an absolute temperature (for example, 30 ° C.) or may be stored as a relative temperature (for example, + 4 ° C.) from the reference temperature. Each set temperature and phase shift amount may be stored as discrete data in a tabular form, or may be stored as an expression expressing the relationship.

  When a temperature setting signal indicating a set temperature is input, the temperature correction information storage unit 31 outputs correction information corresponding to the set temperature indicated by the temperature setting signal to the temperature correction signal generation unit 19.

  The temperature setting signal is output from the computer 7 based on an instruction from the user of the scanning microscope 1 and input through the two-dimensional scanning control circuit 17. Therefore, the user can arbitrarily set the scanning microscope 1 according to the actual usage environment.

  When the correction information indicating the phase shift amount is input, the temperature correction signal generation unit 19 generates a temperature correction signal that causes the phase shift circuit 22 to change the phase of the sine wave signal by the phase shift amount. If the phase shift circuit 22 is an analog circuit including an all-pass filter and a potentiometer, the temperature correction signal is a signal for driving the potentiometer to cause a change in resistance value corresponding to the phase shift amount.

  Thereby, the phase shift circuit 22 changes the phase of the sine wave signal according to the adjustment signal and the temperature correction signal input together with the sine wave signal. More specifically, the phase shift circuit 22 changes the phase of the sine wave signal based on the adjustment signal so that the individual difference deviation at the reference temperature is corrected, and based on the temperature correction signal, the reference temperature The phase of the sine wave signal is changed so that the temperature deviation caused by the difference in the set temperature is corrected. For this reason, the optical scanning position and the electrical sampling position can be matched by matching the set temperature with the use temperature.

In FIG. 2, an example in which the temperature correction information storage unit 31 is included in the sampling clock generation circuit 18 is shown, but the present invention is not limited to this. The temperature correction information storage unit 31 only needs to be able to output correction information to the temperature correction signal generation unit 19, and may be provided outside the sampling clock generation circuit 18, for example, inside the computer 7.
Next, image degradation due to a shift between the optical scanning position and the electrical sampling position will be described.

  FIG. 4 is a diagram for explaining an example of image degradation due to a shift between the optical scanning position and the electrical sampling position. FIG. 4A shows a sample 32 having a pattern in which straight lines are arranged at equal intervals. FIGS. 4B, 4 </ b> D, and 4 </ b> F each illustrate an image of the sample 32 generated by the scanning microscope 1. FIGS. 4C, 4E, and 4G are diagrams showing differences between the images generated in FIGS. 4B, 4D, and 4F and actual samples, respectively. 4C, 4E, and 4G, the vertical axis indicates the difference between the linear interval on the sample and the linear interval on the image, and the horizontal axis indicates the two adjacent straight lines for measuring the interval. The position in the X direction is shown. 4B, 4D, and 4F are images generated in a state where the sample is arranged so that the straight line of the sample and the X direction of the scanning unit are orthogonal to each other.

  In the image 32a illustrated in FIG. 4B, the pattern of the sample 32 is reproduced almost accurately. That is, the image 32a is an image when the optical scanning position matches the electrical sampling position. In this case, as illustrated in FIG. 4C, the linear interval is maintained substantially constant with respect to the X direction.

  On the other hand, in the image 32b illustrated in FIG. 4D and the image 32c illustrated in FIG. 4F, the straight line density gradually increases or decreases in the X direction. That is, the image 32b and the image 32c are images in which the optical scanning position and the electrical sampling position do not coincide with each other and thus are displaced or distorted. In this case, as illustrated in FIG. 4E and FIG. 4G, the straight line interval is not constant in the X direction and changes approximately linearly.

  The image degradation due to such a linear linear interval change is the most typical image degradation of a scanning microscope, and the deviation between the optical scanning position and the electrical sampling position is caused by the phase deviation of the sampling clock signal. It occurs when it is caused.

Therefore, by adjusting the phase shift amount in the phase shift circuit 22 and changing the phase of the sampling clock signal, it is possible to correct the deviation between the optical scanning position and the electrical sampling position.
A procedure for collecting correction information stored in the temperature correction information storage unit 31 will be described.

  First, a sample 32 having a pattern in which straight lines as illustrated in FIG. 4A are arranged at equal intervals is prepared. And the scanning microscope 1 which has arrange | positioned the sample 32 so that a straight line and the X direction may cross orthogonally is accommodated in a thermostat.

Next, the assumed reference temperature and temperature range of the usage environment are determined. In addition, the minimum temperature change amount that causes a recognizable image shift or distortion is determined as a temperature change unit. For example, the reference temperature is 25 ° C., the temperature range is 22 ° C. to 28 ° C., and the temperature change unit is 1 ° C.
After the above preparatory work is completed, the correction information collection procedure illustrated in FIG. 5 is started. FIG. 5 is a flowchart showing a correction information collection procedure.
In step S1, the thermostatic chamber in which the scanning microscope 1 is housed is controlled, and the temperature of the thermostatic chamber (hereinafter referred to as environmental temperature) is changed to the reference temperature.

Next, in step S2, an image of the sample is generated in a state where the environmental temperature matches the reference temperature. Then, the amount of phase shift in the phase shift circuit 22 is adjusted while viewing the generated image, and the image shift and distortion are corrected. In this case, the allowable linear interval error is, for example, 1% of the linear interval on the sample.
In step S3, the signal input to the phase shift circuit 22 in step S2 is set as an adjustment signal.
In step S4, the thermostat is controlled again, and the environmental temperature is changed to the lowest temperature in the temperature range.

  In step S5, an image of the sample is generated in a state where the environmental temperature matches the lowest temperature in the temperature range. Then, the amount of phase shift in the phase shift circuit 22 is adjusted while viewing the generated image, and the image shift and distortion are corrected. In this case, the allowable linear interval error is, for example, 1% of the linear interval on the sample.

In step S6, the phase shift amount in step S5 is temporarily stored in the temperature correction information storage unit 31 in association with the environmental temperature and stored. Note that the phase shift amount to be stored is the difference between the total phase shift amount in the phase shift circuit 22 and the phase shift amount based on the adjustment signal.
In step S7, it is determined whether or not the phase shift amount for the entire temperature range has been acquired. If the phase shift amount for the entire temperature range has not been acquired, the process proceeds to step S8.

  If the phase shift amount for the entire temperature range is acquired, the process proceeds to step S4, where the environmental temperature of the scanning microscope 1 is changed by approximately the temperature change unit. Thereafter, step S4 to step S7 are repeated until the phase shift amount for the entire temperature range is acquired.

  In step S8, an interpolation formula is calculated from the environmental temperature stored in the temperature correction information storage unit 31 by the processing up to step S7 and the phase shift amount corresponding to the temperature. Further, the phase shift amount for each 1 ° C. (temperature change unit) within the temperature range is calculated from the calculated interpolation formula.

  FIG. 6 is a diagram for explaining a method of calculating the interpolation formula and the amount of phase shift. FIG. 6A shows the environmental temperature acquired in step S6 and the phase shift amount corresponding to the temperature. FIG. 6B shows an interpolation formula calculated from the environmental temperature acquired in step S6 and the phase shift amount corresponding to the temperature. FIG. 6C shows the amount of phase shift calculated from the interpolation equation. Thus, by calculating the interpolation equation and calculating the phase shift amount again, the phase shift amount for each temperature change unit can be obtained accurately.

  In step S9, the interpolation equation calculated in step S8 and the phase shift amount for each temperature are stored and stored in the temperature correction information storage unit 31 as correction information. By storing the interpolation formula together with the phase shift amount for each temperature, the phase shift amount can be calculated from the interpolation formula even when the user specifies a set temperature different from the temperature calculated in step S8. Can be output.

  Steps S8 and 9 can be omitted. In this case, the phase shift amount stored in step S6 is used as correction information. In addition, in the calculation of the interpolation formula in step S8, any interpolation method can be adopted. In FIG. 6, an example of linear interpolation is shown, but the present invention is not limited to this.

  Further, in the correction information collection procedure illustrated in FIG. 5, the processing after step S <b> 4 may not be performed for each scanning microscope. If the scanning microscope has the same configuration, the change in characteristics depending on temperature is almost the same. Therefore, correction information is collected only once for each scanning microscope configuration, and the collected correction information is scanned by the same configuration. You may use in common with a type microscope.

  As described above, in the scanning microscope 1 according to the present embodiment, the user changes the set temperature of the scanning microscope 1 according to the temperature of the usage environment, so that the optical scanning position and the electrical sampling position depending on the temperature are changed. The deviation can be corrected. As a result, it is possible to generate an image in which image distortion and displacement are suppressed. For this reason, since the scanning microscope 1 can be normally operated in a wide temperature range, the restriction on the use environment of the scanning microscope 1 can be relaxed.

  In addition, since the scanning microscope 1 according to the present embodiment changes the phase of the signal according to the set temperature instructed by the user, the image distortion is prevented without making the structure and control of the scanning microscope 1 excessively complicated. An image in which the deviation is suppressed can be generated.

The scanning microscope according to the present embodiment is different from the scanning microscope 1 according to the first embodiment in that the sampling clock generation circuit includes a digital circuit instead of the input signal generation unit 21 and the phase shift circuit 22. ing. Other configurations are the same as those of the scanning microscope 1.
FIG. 7 is a diagram illustrating a configuration of a digital circuit used in the sampling clock generation circuit according to the present embodiment.

  The digital circuit 33 illustrated in FIG. 7A includes a phase comparator 34, a loop filter 35, a VCO unit 36, an address counter 37, a ROM 38, and a D / A converter 39. ing.

  First, the VCO unit 36 runs and outputs a clock signal having a constant frequency to the address counter 37. The address counter 37 is a counter whose value is reset every time it counts a predetermined number of times. In addition, a synchronization signal is output to the phase comparator 34 at a timing when the counted value becomes a certain value within a predetermined number of times (hereinafter referred to as an address comparator value).

  The phase comparator 34 compares the phase of the scanning synchronization signal input from the two-dimensional scanning control circuit 17 and the synchronization signal input from the address counter 37. Then, the phase error signal is output to the loop filter 35.

  The loop filter 35 changes the voltage output to the VCO unit 36 according to the phase error signal. As a result, the frequency of the clock signal output from the VCO unit 36 changes.

In this way, by feeding back the output from the VCO unit 36, the cycle of the scanning synchronization signal and the cycle in which the predetermined number of clock signals set in the address counter 37 are output from the VCO unit 36 coincide.
The address counter 37 outputs a count value to the ROM 38 every time the clock signal is counted.

As illustrated in FIG. 7B, the ROM 38 stores sine wave data in which a sine wave shape is reproduced by a number of data equal to the predetermined number of times set in the address counter 37. The ROM 38 outputs sine wave data corresponding to the input count value (address) to the D / A converter 39.
The D / A converter 39 converts the sine wave data that is a digital signal into a sine wave signal that is an analog signal, and outputs the sine wave signal to the position feedback unit 23 (not shown).

  Thereby, the digital circuit 33 can output a sine wave signal having the same period as the scanning synchronization signal, like the input signal generation unit 21 and the phase shift circuit 22 according to the first embodiment.

  In the first embodiment, the number of sampling clock signals per cycle performed by the input signal generation unit 21 is adjusted by the D / A converter 39 in the present embodiment. Specifically, this is performed by changing the amplitude of the sine wave signal output from the D / A converter 39 in accordance with the signal output from the loop filter 30. This can be realized by using a multiplication type D / A converter having a gain adjustment function using a reference voltage terminal or the like as the D / A converter 39.

  In the first embodiment, the phase change of the sine wave signal corresponding to the temperature correction signal and the adjustment signal performed by the phase shift circuit 22 is performed by the function of the address counter 37 in the present embodiment. Specifically, it is performed by changing the timing at which the synchronization signal is output to the phase comparator 34 by adjusting the address comparator value of the address counter 37 in accordance with the temperature correction signal and the adjustment signal.

  FIG. 8 is a diagram for explaining a difference between characteristics of a phase shift circuit configured as an analog circuit and characteristics of a phase shift circuit configured as a digital circuit. FIGS. 8A and 8B show characteristics of a phase shift circuit configured as an analog circuit and a digital circuit, respectively. 8A and 8B, the horizontal axis indicates the amount of phase shift to be changed by the phase shift circuit, and the vertical axis indicates the adjustment set value input to the phase shift circuit in order to realize the phase shift amount. It is. If this set value is configured as an analog circuit, for example, it is a resistance value that determines the time constant of the all-pass filter.

  As illustrated in FIG. 8A, when the phase shift circuit is configured as an analog circuit, the response of the phase shift amount to the set value is degraded as the phase shift amount is increased. That is, the set value and the amount of phase shift are not in a linear relationship. For this reason, the set value set in the phase shift circuit needs to be calculated in accordance with the characteristics of the analog circuit.

  On the other hand, as illustrated in FIG. 8B, when the phase shift circuit is configured as a digital circuit, the set value and the phase shift amount are in a linear relationship. For this reason, the set value set in the phase shift circuit can be easily calculated from the linear relationship with the phase shift amount. For this reason, the structure of the temperature correction signal generation part 19 which produces | generates a temperature correction signal based on the phase shift amount input from the temperature correction information storage part 31 can be simplified.

  In the case of a digital circuit, the phase can be changed from an arbitrary amount of phase shift, that is, from 0 ° to 360 °. On the other hand, in the case of an analog circuit using an all-pass filter, the phase can be changed only up to 180 °.

  As described above, the scanning microscope according to the present embodiment can obtain the same effects as those of the first embodiment. In this embodiment, since the phase shift circuit is configured as a digital circuit, the amount of phase shift is not limited, and the configuration of the temperature correction signal generation unit 19 can be simplified.

The scanning microscope according to the present embodiment is the same as the scanning microscope 1 according to the first embodiment except for the configuration of the sampling clock generation circuit.
FIG. 9 is a diagram illustrating the configuration of the sampling clock generation circuit according to the present embodiment.

  A sampling clock generation circuit 40 illustrated in FIG. 9 includes an input signal generation unit 21, a phase shift circuit 22, a position feedback unit 23, an absolute value circuit unit 41, a VCO unit 25, a counter 26, a D / The A converter 27, the comparator 28, the phase comparison unit 29, the loop filter 30, the temperature correction information storage unit 42, and the temperature correction signal generation unit 43 are configured.

  The sampling clock generation circuit 40 is different from the sampling clock generation circuit 18 according to the first embodiment in that the adjustment signal and the temperature correction signal are output to the phase shift circuit 22 and the absolute value circuit unit 41. Accordingly, in the sampling clock generation circuit 40, the absolute value circuit unit 41 is replaced with the temperature correction information storage unit 42 instead of the absolute value circuit unit 24, the temperature correction information storage unit 42 is replaced with the temperature correction information generation unit 19. Instead, a temperature correction signal generator 43 is included.

  FIG. 10 is a diagram for explaining an example of image degradation due to a shift between the optical scanning position and the electrical sampling position. FIG. 10A shows a sample 32 having a pattern in which straight lines are arranged at equal intervals. FIGS. 10B, 10 </ b> D, and 10 </ b> F illustrate images of the sample 32 generated by the scanning microscope 1. FIGS. 10C, 10E, and 10G are views showing the difference between the image generated in FIGS. 10B, 10D, and 10F and the actual sample, respectively. In addition, the vertical axis | shaft of FIG.10 (c), (e), (g) shows the difference of the linear space | interval on a sample and the linear space | interval on an image, and a horizontal axis | shaft of two adjacent straight lines which measure space | interval. The position in the X direction is shown. 10B, 10D, and 10F are images generated in a state where the sample is arranged so that the straight line of the sample and the X direction of the scanning unit are orthogonal to each other.

  In the image 32d illustrated in FIG. 10B, the pattern of the sample 32 is reproduced almost accurately. That is, the image 32d is an image when the optical scanning position matches the electrical sampling position. In this case, as illustrated in FIG. 10C, the linear interval is maintained substantially constant with respect to the X direction.

  On the other hand, in the image 32e illustrated in FIG. 10D and the image 32f illustrated in FIG. 10F, the density of straight lines gradually increases or decreases from the vicinity of the center of the image toward both ends of the image. Yes. That is, the image 32d and the image 32f are images in which the optical scanning position and the electrical sampling position do not coincide with each other and thus are displaced or distorted. In this case, as illustrated in FIGS. 10E and 10G, the linear interval is not constant with respect to the X direction, but changes nonlinearly, and shows a bowl-like shape as a whole.

  The deterioration of the image due to such a non-linear change in the linear interval is not appropriate for the relationship between the maximum scanning speed and the minimum scanning speed within one period of the scanning means and the maximum frequency and the minimum frequency of the sampling clock signal, and there is an error. It happens because there is.

For this reason, the offset voltage is applied to the sine wave signal that has been subjected to full-wave rectification processing, and the balance between the highest frequency and the lowest frequency of the sampling clock signal is changed, thereby correcting the deviation between the optical scanning position and the electrical sampling position. be able to.
FIG. 11 is a diagram for explaining the configuration and operation of the absolute value circuit unit 41 included in the sampling clock generation circuit 40 according to this embodiment.

As illustrated in FIG. 11A, the absolute value circuit unit 41 includes a full-wave rectification unit 44 and an offset unit 45 that applies an offset voltage to the full-wave rectified signal. .
The sine wave signal input from the phase shift circuit 22 is full-wave rectified by the full-wave rectification unit 44 and output to the offset unit 45.

  The offset unit 45 functions as a gain adjustment unit that changes the amplitude of the sine wave signal, and applies an offset voltage corresponding to the adjustment signal and the temperature correction signal to the full-wave rectified sine wave signal. To change the amplitude.

  When the offset voltage is applied, as illustrated in FIG. 11B, the voltage of the sine wave signal temporarily becomes higher than the signal 41a before the application of the offset voltage. That is, both the highest frequency and the lowest frequency of the sampling clock signal are increased (see signal 41b). However, the number of sampling clock signals generated per period is fixed by the PLL operation of the sampling clock generation circuit 40 that feeds back the output from the VCO unit 25 to the input signal generation unit 21. For this reason, the amplitude of the sine wave signal is reduced and the maximum frequency of the sampling clock signal is lowered, so that the control is stabilized in a state where the balance between the maximum frequency and the minimum frequency is changed (see signal 41c).

  For this reason, the balance between the highest frequency and the lowest frequency can be changed by adjusting the offset voltage using the adjustment signal and the temperature correction signal, thereby correcting the deviation between the optical scanning position and the electrical sampling position. Can do.

  As described above, the scanning microscope according to the present embodiment can achieve the same effects as those of the scanning microscope 1 according to the first embodiment. Further, in this embodiment, since the temperature correction signal is output to the input signal generation unit 21 and the absolute value circuit unit 41, image degradation due to a change in linear linear interval as illustrated in FIG. 10, it is possible to cope with any image degradation caused by a non-linear change in linear spacing.

  Also, the scanning microscope according to the present embodiment may be configured using the digital circuit 33 instead of the input signal generation unit 21 and the phase shift circuit 22, as in the scanning microscope according to the second embodiment. Thereby, the same effect as the scanning microscope according to the second embodiment can be obtained.

  In the present embodiment, the correction information stored in the temperature correction information storage unit 42 is a phase shift amount and an offset voltage. Then, the temperature correction signal generation unit 43 outputs a signal for changing the phase of the sine wave signal by the amount of phase shift to the phase shift circuit 22 and outputs a signal for applying an offset voltage to the full-wave rectified signal. 41 is output. These correction information can be collected by the same procedure as in the first embodiment.

  FIG. 12 is a diagram illustrating the configuration of the scanning microscope according to the present embodiment.

  The scanning microscope 46 illustrated in FIG. 12 relates to the first embodiment in that the sampling clock generation circuit 18 includes a temperature sensor 47 and a temperature change unit 48 that changes the temperature of the sampling clock generation circuit 18. Different from the scanning microscope 1. Other configurations are the same as those of the scanning microscope 1 according to the first embodiment.

  FIG. 12 shows an example in which the temperature sensor 47 is arranged in the sampling clock generation circuit 18. This is because, in general, the temperature change of the sampling clock generation circuit 18 has the greatest influence on the image, but the arrangement of the temperature sensor 47 is not particularly limited to this.

  The temperature changing section 48 may change the temperature in a local range including the sampling clock generation circuit 18, and changes the temperature of the sampling clock generation circuit 18 by changing the internal atmosphere temperature of the microscope body 2. Things can be used. The temperature changing unit 48 may be, for example, a temperature control element such as a Peltier element, a heater, a cooler, or a circuit that controls the operation of the air cooling fan.

The temperature changing unit 48 is mainly used when collecting correction information for each temperature. In the scanning microscope 46 according to the present embodiment, since the temperature of the sampling clock generation circuit 18 is changed using the temperature changing unit 48, a constant temperature bath that houses the scanning microscope 46 and changes the environmental temperature is not always necessary. Absent.
Further, more accurate correction information can be obtained as compared with the case where the environmental temperature is changed using a thermostatic chamber as exemplified in the first embodiment.

  When the environmental temperature is changed by the thermostatic bath, there is a time lag between the temperature change of the thermostatic bath and the temperature change of a portion having a large influence on the image such as a sampling clock generation circuit. For this reason, when the correction information is acquired based on the temperature of the thermostatic chamber, the correction information is acquired before the temperature of the sampling clock generation circuit or the like is stabilized, and as a result, accurate correction information may not be obtained. .

  In addition, in order to obtain accurate correction information, it is necessary to wait until the temperature of the sampling clock generation circuit or the like changes due to the temperature change of the thermostat and becomes stable. In this case, since it is difficult to accurately grasp the time until a stable state is reached, it takes a longer time than necessary. Therefore, it takes time to obtain correction information.

  In contrast, in the case of the scanning microscope 46 according to the present embodiment, the temperature of the sampling clock generation circuit 18 is changed by the temperature changing unit 48 while the temperature of the sampling clock generation circuit 18 is measured by the temperature sensor 47. Since the object whose temperature is to be measured matches the object whose temperature is to be changed, no time lag occurs. For this reason, more accurate correction information can be acquired in the minimum necessary time.

  The correction information collection procedure is the same as the collection procedure of the first embodiment. However, when a circuit for controlling the operation of the air cooling fan or the like is used as the temperature changing unit 48, an image is generated and corrected at regular intervals while changing the environmental temperature by the temperature changing unit 48, and the phase shift amount used for the correction May be stored in the temperature correction information storage unit 42 together with the environmental temperature at that time. Even when correction information is collected in this way, it is possible to obtain correction information for each temperature that differs by a temperature change unit by obtaining an interpolation formula from the collected information.

  In the scanning microscope 46 according to this embodiment, it is necessary to set the temperature of the sampling clock generation circuit 18 as the set temperature of the scanning microscope 46, not the internal atmosphere temperature of the scanning microscope 46. Therefore, the temperature of the sampling clock generation circuit 18 detected by the temperature sensor 47 may be displayed on the monitor 8 so that the user can check the temperature of the sampling clock generation circuit 18. Thereby, the user can set an appropriate set temperature.

  Further, the scanning microscope 46 may automatically set the temperature detected by the temperature sensor 47 to the set temperature instead of the user setting the set temperature. In this case, if the set temperature is changed following a slight temperature change, the set temperature of the scanning microscope 46 is frequently changed, and the performance of the scanning microscope 46 may be adversely affected. For this reason, a dead zone may be provided in advance so that the set temperature is not changed within a range in which image degradation due to a temperature change is allowable. The width of the dead zone may be the same as the temperature change unit, for example.

  Further, when the correction information corresponding to the temperature detected by the temperature sensor 47 is not stored in the temperature correction information storage unit 31, the fact may be displayed on the monitor 8 to notify the user. In this case, it may be selected whether the correction information is generated and used from a previously calculated interpolation formula, or whether the user manually corrects the image.

  When the image is manually corrected by the user, the information at that time may be added to the temperature correction information storage unit 31 as correction information. In this case, it is desirable to calculate the interpolation formula anew including the added correction information. This improves the accuracy of the interpolation formula.

  As described above, the scanning microscope 46 according to the present embodiment can obtain the same effects as those of the scanning microscope 1 according to the first embodiment. In the scanning microscope 46 according to the present embodiment, by providing the temperature sensor 47, it is possible to improve the accuracy of the correction information, shorten the collection time of the correction information, and automate the setting of the temperature setting signal. it can. Further, by providing the temperature changing section 48, a large facility such as a thermostatic bath can be omitted. Further, by notifying the presence / absence of the correction information through the monitor 8 and providing the user with an opportunity to acquire the correction information, the accuracy of the correction information deteriorates due to unlimited interpolation (including extrapolation). Can be suppressed.

  Further, the scanning microscope 46 according to the present embodiment may be configured using a digital circuit 33 instead of the input signal generation unit 21 and the phase shift circuit 22, as in the scanning microscope according to the second embodiment. . Thereby, the same effect as the scanning microscope according to the second embodiment can be obtained. In addition, the scanning microscope 46 according to the present embodiment may input a temperature correction signal to the phase shift circuit 22 and the absolute value circuit unit, similarly to the scanning microscope according to the third embodiment. Thereby, the same effect as the scanning microscope according to the third embodiment can be obtained.

DESCRIPTION OF SYMBOLS 1,46 ... Scanning microscope, 2 ... Microscope main body, 3, 32 ... Sample, 4 ... Stage, 5 ... Objective lens, 6 ... Revolver, 7 ... Computer, DESCRIPTION OF SYMBOLS 8 ... Monitor, 9 ... Laser light source, 10, 13 ... Mirror, 11 ... Half mirror, 12 ... Two-dimensional scanning part, 14 ... Lens, 15 ... Confocal stop , 16 ... photodetector, 17 ... two-dimensional scanning control circuit, 18, 40 ... sampling clock generating circuit, 19, 43 ... temperature correction signal generating unit, 20 ... Z-axis scanning unit , 21 ... input signal generation unit, 22 ... phase shift circuit, 23 ... position feedback unit, 24, 41 ... absolute value circuit part, 25, 36 ... VCO part, 26 ... Counter, 27, 39 ... D / A converter, 28 ... Comparator, 29 ... Phase ratio Part, 30, 35 ... loop filter, 31, 42 ... temperature correction information storage part, 32a, 32b, 32c, 32d, 32e, 32f ... image, 33 ... digital circuit, 34 ... Phase comparator, 37 ... Address counter, 38 ... ROM, 41a, 41b, 41c ... Signal, 44 ... Full-wave rectification unit, 45 ... Offset unit, 47 ... Temperature sensor, 48 ... Temperature change section

Claims (14)

Scanning means for moving the collection position of the illumination light applied to the sample by sinusoidal vibration;
Light detection means for detecting observation light obtained by irradiating the sample with the illumination light, and converting the observation light into an image detection signal that is an electrical signal;
Sampling clock generating means for generating a sampling clock signal indicating timing for sampling the image detection signal from a scanning synchronization signal having a period equal to the period of the sine wave vibration;
Sampling the image detection signal based on the sampling clock signal, and generating an image of the sample, and
The sampling clock generation means includes
Temperature correction means for generating a temperature correction signal for correcting the sampling clock signal according to a set temperature;
Temperature correction information storage means for storing correction information corresponding to the set temperature;
A frequency variable oscillating means for inputting a sine wave signal obtained by converting the scanning synchronization signal and outputting the sampling clock signal;
Phase shifting means for changing the phase of the sine wave signal;
Gain adjusting means for changing the amplitude of the sine wave signal,
The temperature correction unit receives the correction information corresponding to the set temperature from the temperature correction information storage unit, and changes the amplitude of the sine wave signal to the gain adjustment unit according to the set temperature. A scanning microscope characterized by inputting a temperature correction signal. Scanning means for moving the collection position of the illumination light applied to the sample by sinusoidal vibration;
Light detection means for detecting observation light obtained by irradiating the sample with the illumination light, and converting the observation light into an image detection signal that is an electrical signal;
Sampling clock generating means for generating a sampling clock signal indicating timing for sampling the image detection signal from a scanning synchronization signal having a period equal to the period of the sine wave vibration;
Sampling the image detection signal based on the sampling clock signal, and generating an image of the sample, and
The sampling clock generation means includes
Temperature correction means for generating a temperature correction signal for correcting the sampling clock signal according to a set temperature;
Temperature correction information storage means for storing correction information corresponding to the set temperature;
A frequency variable oscillating means that receives the sine wave signal converted from the scanning synchronization signal and outputs the sampling clock signal;
The temperature correction unit receives the correction information corresponding to the set temperature from the temperature correction information storage unit, and determines the amplitude of the sine wave signal input to the frequency variable oscillation unit according to the set temperature. A scanning microscope characterized by being changed. The scanning microscope according to claim 1 ,
The scanning microscope according to claim 1, wherein the temperature correction means inputs the temperature correction signal for changing the phase of the sine wave signal to the phase shift means in accordance with the set temperature. The scanning microscope according to claim 2,
The scanning microscope characterized in that the temperature correction means changes the phase of the sine wave signal input to the variable frequency oscillation means in accordance with the set temperature. The scanning microscope according to claim 2,
The sampling clock generation means further includes:
Phase shifting means for changing the phase of the sine wave signal;
And a gain adjusting means for changing the amplitude of the sine wave signal. The scanning microscope according to claim 5 ,
The scanning microscope according to claim 1, wherein the temperature correction means inputs the temperature correction signal for changing the phase of the sine wave signal to the phase shift means in accordance with the set temperature. In the scanning microscope according to claim 5 or 6 ,
The scanning microscope characterized in that the temperature correction means inputs the temperature correction signal for changing the amplitude of the sine wave signal to the gain adjustment means in accordance with the set temperature. In scanning microscope according to any one of claims 1 to 7,
The scanning microscope further includes a temperature sensor. The scanning microscope according to claim 8, wherein
The scanning microscope according to claim 1, wherein the temperature sensor detects a temperature of the sampling clock generation means. The scanning microscope according to claim 9, wherein
The scanning microscope according to claim 1, wherein the temperature correction unit generates the temperature correction signal for correcting the sampling clock signal in accordance with the temperature detected by the temperature sensor. The scanning microscope according to claim 9, wherein
Furthermore, the scanning microscope characterized by including a notification means for notifying that the correction information corresponding to the temperature detected by the temperature sensor is not stored in the temperature correction information storage means. The scanning microscope according to any one of claims 9 to 11,
The scanning microscope further includes temperature changing means for changing the temperature of the sampling clock generating means. The scanning microscope according to any one of claims 1 to 12,
The scanning microscope according to claim 1, wherein an adjustment signal for adjusting the sampling clock signal is input to the sampling clock generation means in accordance with individual characteristics of the scanning microscope. The scanning microscope according to any one of claims 1 to 13,
Furthermore, the scanning microscope characterized by including the laser light source which inject | emits a laser beam as said illumination light.

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