JP2000267011A - Scanning laser microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To correct the phases of an optical scanning position and a sampling clock for every line in real time, to realize it with a simple constitution and to obtain an image without distortion and deterioration. SOLUTION: A laser light beam Q is used for scanning a sample 4 by a deflection means 2. Besides, reflected light or fluorescence R from the sample 4 is detected and the image signal thereof is obtained by a photodetector 6. Then, the image processing of the image signal obtained by the photodetector 6 is executed by an image processing system 7 according to the sampling clock (p) and a trigger pattern (k). In this case, the scanning position of the beam Q on the sample 4 by the deflection means 2 is detected by a sensor 8. Based on the scanning position of the beam Q detected by the sensor 8, the phases of the clock (p) given to the processing system 7 and the scanning position of the beam Q are made to coincide with each other by a phase correction means 9.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a scanning laser microscope that scans a sample with a laser beam, detects reflected light or fluorescence from the sample, and obtains an image of the sample by image processing.

[0002]

2. Description of the Related Art In a scanning laser microscope, a laser beam, which is a point light source, is applied to an X-axis and a Y-axis of a sample through an objective lens.
Irradiation is performed while scanning in the axial direction, and reflected light or fluorescence from the sample is detected again by a detector via an optical system such as an objective lens to obtain density information of a two-dimensional image. Is displayed on an image output device such as a CRT monitor or a color printer as a distribution of bright spots in correspondence with the X-Y scanning position, thereby forming an image for observation.

Further, by providing a stop having a diameter approximately equal to the diffraction limit of the illumination light or the light to be measured at a position conjugate with the sample of the detection optical system, it is possible to detect only the information of the focused plane. The result is a confocal scanning laser microscope.

In such a scanning laser microscope, a galvanometer mirror is mainly used as means for two-dimensionally scanning a laser beam from a light source. When using a galvanometer mirror as two-dimensional scanning means,
For example, as disclosed in Japanese Patent Publication No. Hei 6-27905, a driving signal waveform is a triangular wave or a sawtooth wave, and data is sampled by a sampling clock having a constant period in a portion where the displacement of the mirror moves linearly with time. In some cases, images without distortion were captured.

[0005] In addition, rather than driving a galvanometer mirror with a drive signal having a high-frequency component at the apex of a peak or a valley in a waveform, such as a triangular wave or a sawtooth wave, a drive signal having a single frequency, For example, it is also known that an image can be acquired at high speed while suppressing distortion of an obtained image by modulating the period of the sampling clock so as to compensate for the fluctuation of the scanning speed of the mirror by making the mirror a sinusoidal wave. is there.

The following two methods are generally known as methods for modulating and generating the period of the sampling clock so as to compensate for the above-mentioned fluctuation in the scanning speed of the mirror.

The first method is disclosed in, for example, Japanese Patent Application Laid-Open No. 5-2135.
JP-A-5-136954, JP-A-5-136954
For example, a sample is scanned by a laser beam, and at the same time, a linear scale pattern is scanned, and a sampling clock is generated from the intensity of reflected light or transmitted light from the pattern. This is the method that was used.

The second method is a method in which a sampling clock pattern is created on a memory so that data can be sampled at a linear scanning position for a sine wave of a drive signal waveform.

[0009]

However, these compensation methods have the following problems.

That is, in the first method, it is necessary to install an optical system for scanning a linear scale, separately from the optical system for scanning the sample, and further scan the linear scale in the optical system for scanning the sample. However, there is a problem that the loss of the light amount is large because the optical system is installed.

In the second method, there is a problem that a phase delay occurs between the drive signal waveform and the actual scanning position of the mirror due to the phase characteristics of the mirror itself and the drive circuit.

As shown in FIG. 10, the phase delay changes with the change of the driving frequency, and the amount of the phase delay changes with the change of the amplitude.

If the phase delay amount between the drive signal waveform and the sampling clock is not changed appropriately for such a change in the phase delay, there is a problem that an obtained image is distorted.

Further, when data is sampled in both the forward path and the return path, since the timing of the synchronization signal indicating the sampling start position must be finely adjusted, it is extremely difficult to sample the same position in the forward path and the return path. There was also a problem that it was difficult.

Also, a phase delay between a drive signal waveform arbitrarily generated and a sampling clock, for example, a phase shift between a scanning position of a laser beam and a sampling clock generated due to factors such as electrical, mechanical characteristics, and environment. However, when the delay or the phase changes for each line due to the change in the delay amount due to the temperature or the characteristics of the deflecting means that scans the sample with the laser beam, it cannot be dealt with.

Accordingly, the present invention provides a scanning laser microscope which performs real-time correction of the optical scanning position and the phase of a sampling pattern line by line in a real-time manner and realizes an image free from distortion and deterioration. The purpose is to provide.

[0017]

According to the first aspect of the present invention, a deflecting means for scanning a sample with a laser beam, a light detecting means for detecting light from the sample and obtaining an image signal thereof, Image processing means for performing image processing on an image signal obtained by the detection means in accordance with a sampling clock and a trigger pattern; scanning position detection means for detecting a scanning position of the laser beam on the sample by the deflection means; and detection by the scanning position detection means A scanning laser microscope comprising: a sampling clock provided to an image processing unit based on the scanning position of the laser beam; and a phase correction unit for matching the phase of the scanning position of the laser beam.

According to a second aspect of the present invention, in the scanning laser microscope according to the first aspect, the phase correction means stores a sampling clock and a trigger pattern, and scans the scanning position of the laser light detected by the scanning position detection means. It has a function of detecting a phase difference between the trigger pattern and the trigger pattern, and providing a sampling clock to the image processing means according to the phase difference.

According to a third aspect of the present invention, in the scanning laser microscope according to the first aspect, the phase correction means stores a sampling clock and matches the scanning position of the laser beam detected by the scanning position detection means. As described above, the sampling clock is read and provided to the image processing means.

[0020]

(1) Hereinafter, a first embodiment of the present invention will be described with reference to the drawings.

FIG. 1 is a configuration diagram of a scanning laser microscope.

The light source 1 emits a laser beam Q.
This laser light Q is sent to the deflection means 2. The deflecting means 2 has a function of receiving the sine wave drive signal d from the drive circuit 3 and scanning the sample 4 with the laser light Q in the XY directions. In this case, the drive circuit 3 stores the drive waveform pattern of the sine wave drive signal d as shown in FIG. 2 in the storage means 5 such as a memory, and one cycle of the drive signal d as shown in FIG. The sample 4 is reciprocally scanned on the sample 4. That is, the memory data of the drive signal d is P1,
In periods t1 and t2 between P2, the forward path t1 and the backward path t2 of scanning on the sample 4 are repeated.

The photodetector 6 has a function of detecting reflected light or fluorescence R from the specimen 4 and obtaining an image signal of the reflected light or the fluorescence R. The image signal is sent to an image processing system 7.

The image processing system 7 has a function of inputting an image signal obtained by the photodetector 6, processing the image signal to obtain image data of the specimen 4, and displaying the image data on a monitor or the like as a microscope image. are doing. In this case, the image processing system 7 processes the image signal obtained by the photodetector 6 according to the sampling clock p for performing the image processing.

In the storage means 5 of the drive circuit 3, in addition to the sine wave drive signal d, a sampling clock p for performing image processing and the phase of the sampling clock p coincide with the phase of the scanning position of the laser beam. Therefore, a trigger pattern k for detecting the phase difference is stored.
The sampling clock p and the trigger pattern k are written at the same address so as to correspond to the phase of the drive signal d.
And the trigger pattern k can be obtained.

On the other hand, the sensor 8 detects the optical position of the laser beam on the specimen 4 and sends an electric signal of the optical scanning position to the phase correcting means 9.

The phase correction means 9 receives the sampling clock p and the trigger pattern k stored in the storage means 5 of the drive circuit 3 and stores them in a storage device (not shown). A device having a function of detecting a phase difference between a scanning position of a laser beam detected by an electric signal of a position and a trigger pattern, reading the sampling clock P whose phase has been corrected according to the phase difference from a storage device, and providing the read clock to an image processing system 7. It is. Note that a storage device serving as a temporary buffer, such as a FIFO, is preferable as the storage device of the phase correction unit 9.

Next, the operation of the scanning laser microscope configured as described above will be described.

The laser light Q emitted from the light source 1 is
The light is incident on the deflection means 2.

The deflecting means 2 operates when a driving signal d is supplied from a driving circuit 3, and moves the position of the laser beam Q applied to the sample 4 to thereby generate the driving signal d.
Is scanned in a desired area corresponding to. This drive signal d is
As described above, the storage means 5 such as a memory in the drive circuit 3
Is stored as a waveform pattern, and when scanning starts, a periodic drive signal is obtained by repeatedly reading out the pattern. The laser beam incident on the deflecting means 2 is scanned by the deflecting means 2 by the drive signal d output from the drive circuit 3 and guided onto the sample 4 as described above.

On the other hand, the sampling clock p and the trigger pattern k are read from the storage means 5 simultaneously with the drive signal d, and these sampling clock p and trigger pattern k are sent to the phase correction means 9.

The phase correcting means 9 corrects the phase of the sampling clock p output from the drive circuit 3 so as to match the phase of the optical scanning position of the laser beam Q for scanning the sample 4. That is, since the sampling clock p stored in the storage means 5 is simultaneously read from the output of the driving circuit 3, the driving signal d, the sampling clock p and the trigger pattern k are in phase, but the driving signal d and the sample At the optical scanning position of the laser light Q on 4, a phase difference occurs due to electrical and mechanical characteristics and the environment.

As a matter of course, since the sampling clock p and the trigger pattern k also have a phase difference from the optical scanning position of the laser beam Q, the image signal obtained by scanning the sample 4 is processed. Therefore, it is necessary to correct the phase of the sampling clock p.

Therefore, the sensor 8 detects the optical scanning position of the laser beam Q and outputs an electric signal.

The phase correction means 9 receives the electric signal output from the sensor 8 and compares it with the trigger pattern k generated by the drive circuit 3 to detect a phase difference. Then, the phase correction means 9 converts the sampling clock p output by the drive circuit 3 into a temporary buffer such as a FIFO at the time when scanning starts or at the timing of the trigger pattern k output by the drive circuit 3. Are sequentially stored in a storage device.

When the phase correction means 9 receives the electric signal from the sensor 8, the phase correction means 9 starts reading the sampling clock P temporarily stored from the storage device.
To send to. That is, even if the sampling clock p is read from the drive circuit 3, it is not directly sent to the image processing system 7, but is temporarily stored in a storage device such as a FIFO and the optical position at which the sample 4 is scanned. The phase is corrected by reading the signal in accordance with the timing of the signal, that is, the output signal of the sensor 8, and is sent to the image processing system 7.

As a result, the image processing system 7 includes the light detector 6
The sampling signal P corrected to match the phase of the optical scanning position is supplied to the image signal of the sample 4 obtained by the above, and the image processing is performed at a normal timing.

As described above, in the first embodiment, the optical scanning position of the laser beam Q on the sample 4 is detected, and the sampling clock P applied to the image processing system 7 based on the optical scanning position is detected. And the scanning position of the laser light Q are made to coincide with each other, so that the optical scanning position of the laser light Q generated by factors such as electrical, mechanical characteristics, and environment and the sampling clock p for image processing. Can be detected and corrected for the phase difference, and a sampling clock P in phase with the optical scanning position can be immediately obtained. Further, even for a phase shift different for each line, it is possible to perform a correction corresponding to each shift amount, and furthermore, it is possible to realize with a simple configuration.

(2) Next, a second embodiment of the present invention will be described with reference to the drawings. The same parts as those in FIG. 1 are denoted by the same reference numerals, and detailed description thereof will be omitted.

FIG. 4 is a configuration diagram of a scanning laser microscope.

The light source 1 emits a laser beam Q.
The laser light Q is applied to a deflection means 10 in a horizontal direction (X direction).
To the deflection means 11 in the vertical direction (Y direction).
Among these, an X-direction galvanometer mirror is used as the deflecting means 10, and a Y-direction galvanometer mirror is used as the deflecting means 11, and these X-direction galvanometer mirrors 10 and Y
The direction galvanomirror 11 deflects the laser beam Q two-dimensionally in the horizontal and vertical directions. These deflecting means 10 and 11 are driven by an X-direction drive circuit 12 and a Y-direction drive circuit 13, respectively.

On the other hand, the memory read control circuit 15 and the memory 16 are controlled by a command from the CPU 14. The memory read control circuit 15 receives the CLK
It has a function of controlling a read operation of the memory 16 in response to a signal.

The memory 16 has a driving waveform pattern of a sine wave driving signal d of the X-direction driving circuit 12 and the Y-direction driving circuit 13, a sampling clock p and a trigger pattern k.
Is written by the CPU 14.
The drive signal d, the sampling clock p and the trigger pattern k are read out simultaneously, and the drive signal d
Are analog-converted by a D / A converter 17 and sent to the X-direction drive circuit 12 and the Y-direction drive circuit 13, and the sampling clock p and the trigger pattern k are sent to the phase correction circuit 9.

The phase correction circuit 9 receives the sampling clock p and the trigger pattern k stored in the memory 16 and temporarily stores them, and is detected by the electric signal of the optical scanning position output from the sensor 8. Detects the phase difference between the scanning position of the laser beam and the trigger pattern k,
It has a function of starting reading of the sampling clock P temporarily stored according to this phase difference and giving it to the image processing system 7.

FIG. 5 is a specific configuration diagram of the phase correction circuit 9.

The voltage comparison circuit 18 receives the electric signal of the optical scanning position from the sensor 8 and the reference voltage set in the reference voltage register 19, that is, the drive signal d as shown in FIG.
And outputs a low level (L level) when the output signal of the sensor 8 is equal to or lower than the voltage value P1 and equal to or higher than the voltage value P2, and is equal to or lower than the voltage value P1 and equal to or lower than the voltage value P2. Has a function of outputting a high level (H level) when.

The FIFO (first-in-first-out) control circuit 20 starts the write operation to the FIFO 21 in response to the rise of the trigger pattern k read from the memory 16, and starts the sampling clock read from the memory 16. p is sequentially written to the FIFO 21,
In addition, it has a function of reading out the FIFO 21 at the timing when the output signal of the voltage comparison circuit 18 becomes H level, obtaining the sampling clock P with the phase corrected, and sending it to the image processing system 7.

The FIFO 21 has a first-in-
This is a first-out storage device in which the sampling clock p is sequentially written and read out in the writing order.

The sensor 8 detects the optical scanning position of the laser beam Q split by the half mirror 22 disposed between the X-direction galvanometer mirror 10 and the Y-direction galvanometer mirror 11. But this sensor 8
Of the light source 22 for detection as shown in the broken line in the figure.
May be provided, and the laser light output from the light source 22 may be applied to the X-direction galvanometer mirror 10, and the reflected light from the X-direction galvanometer mirror 10 may enter the sensor 8.

Next, the operation of the scanning laser microscope configured as described above will be described.

The CPU 14 writes the drive signal d, the sampling clock p and the trigger pattern k in the memory 16 in advance.

FIG. 6 shows an image of the memory 16 storing the drive signal d of the X-direction galvanometer mirror 10, the sampling clock p and the trigger pattern k. The addresses in the memory 16 are shown in the horizontal direction, and the data values are shown in the vertical direction. This is an example in which reciprocal scanning is performed using a sine wave (sin wave). One line is scanned in the forward path from addresses A1 to A2 to acquire 10 pixels, and one line is scanned in the backward path from A3 to A4. Get the pixels. One cycle of a sine wave shows
The drive signal d of the X galvanometer mirror 10 is a sampling clock p having 20 pulses and a trigger pattern k having two pulses. In the sampling areas A1 to A2 and A3 to A4, the sine wave of the drive signal d does not change linearly, so that the sampling clock is not a fixed cycle. Here, the trigger pattern k is stored in the memory 16 as a pattern indicating the sampling start position and the sampling end position of the line.
That is, a signal in which the data value of the drive signal d rises at the address A1 which becomes P1 on the outward path and the address A3 which becomes P2 on the return path, and falls at the address A2 which becomes P2 on the outward path and at the address A4 which becomes P1 on the return path. It is.

Since these sampling clock p and trigger pattern k can take only two values of H level and L level, 1 bit is assigned to the data of the memory 16 and the H level is set to 1 and the L level is set to 0. Is written. The sine wave is assigned several bits according to the resolution.

When the scanning is started, the CPU 14 outputs the timing at which the scanning is started to the memory read control circuit 15, and in response thereto, the memory read control circuit 1
5 counts the CLK signal with a counter provided therein,
The counting is continued until the scan end timing is input.
The output of the counter is stored in memory 1 as a memory address.
6. Here, the CLK signal is a clock of a uniform cycle for reading out the memory, and is also supplied to the phase correction circuit 9.

The memory 16 includes a memory read control circuit 1
5, the data of the corresponding address, that is, the drive signal d, the sampling clock p, and the trigger pattern k are simultaneously output. Of these, the pattern of the drive signal d is sent to the D / A converter 17 to be converted into an analog signal, sent to the X-direction drive circuit 12 and the Y-direction drive circuit 13, and based on the drive signal d, X, Y The directional galvanometer mirrors 10 and 11 are respectively driven.

At the same time, the sampling clock p and the trigger pattern k read from the memory 16 are sent to the phase correction circuit 9 and are output from the sensor 8 provided for detecting the optical scanning position of the laser light Q. It is compared with the output signal to correct the phase of the sampling clock p so as to match the optical scanning position. The corrected sampling clock P is output to the image processing system 7 and processes the image signal obtained from the photodetector 6.

Here, the operation of the phase correction circuit 16 will be described in detail.

The FIFO control circuit 20 starts the write operation of the FIFO 21 at the rise of the trigger pattern k read from the memory 16 (A1 shown in FIG. 6), and the sampling clock p output from the memory 16
Are sequentially written to the FIFO 21.

On the other hand, the reading of the sampling clock p stored in the FIFO 21 starts upon receiving the output timing of the voltage comparison circuit 18. This voltage comparison circuit 18
Compares the output signal of the sensor 8 with the voltage set in the reference voltage register 19 and outputs the result.

For example, in the reference voltage register 19, FIG.
The voltage values of P1 and P2 in the drive signal d shown in FIG.

The voltage comparison circuit 18 outputs the L level when the output signal from the sensor 8 is lower than the reference voltage P1 and when the output signal is higher than the reference voltage P2, and otherwise outputs the H level.
Output level. Thus, the output of the voltage comparison circuit 18 has the same waveform as the trigger pattern k stored in the memory 16 and is delayed in time.

Referring specifically to FIG. 7, the sine wave and the rectangular wave indicated by the solid line indicate the drive signal d and the trigger pattern k read from the memory 16, and the sine wave indicated by the broken line indicates , The signal indicating the optical scanning position detected by the sensor 8 and the dashed rectangular wave indicate the output signal of the voltage comparison circuit 18 when the circuit is configured as described above.

The drive signal d output from the memory 16 and
The optical scanning position detected by the sensor 8 is delayed due to factors such as the characteristics of the apparatus and the environment, and a phase difference is generated.

Therefore, the trigger pattern k read simultaneously with the drive signal d is used.

First, the FIFO control circuit 20 detects a rising edge of the trigger pattern k and starts a write operation to the FIFO 21. The sampling clock p output from the memory 16 is sequentially written into the FIFO 21.

On the other hand, the reading of the sampling clock p from the FIFO 21 is performed by using the rising edge of the output signal obtained by the voltage comparing circuit 18 based on the output signal of the sensor 8 as the FIFO.
The operation is started by the detection by the O control circuit 20. FI
When the read operation is started, the FO 21 sequentially outputs data from the head of the written sampling clock P. The write operation is continued during that time.

When the falling of the output signal of the voltage comparing circuit 18 is received, the FIFO control circuit 20
O21 is cleared, and both the write operation and the read operation are stopped. Thereafter, the apparatus is in a standby state until the next trigger pattern rises, and this operation is repeated thereafter.

Therefore, the phase difference between the drive signal d output from the memory 16 and the optical scanning position by the sensor 8 is:
A period from the start of writing to the FIFO 21 to the start of reading, that is, the delay amount Δt1 shown in FIG.
, The delay amount Δ
t1 can be absorbed and corrected.

Further, for example, the delay amount of the next line (FIG. 7)
, The delay amount Δt2) of the previous line (the delay amount Δt)
Even in the case different from 1), an arbitrary delay amount can be easily corrected by using the trigger pattern k and the output signal of the voltage comparison circuit 18.

However, the image processing system 7 processes the image signal from the photodetector 6 using the sampling clock P for image processing that matches the optical scanning position of the laser beam Q on the sample 4. To obtain an observation image of the specimen 4.

As described above, according to the second embodiment, similarly to the first embodiment, the optical scanning position of the laser beam Q generated by factors such as electrical, mechanical characteristics, and environment. And the sampling clock p for image processing can be detected and corrected, and a sampling clock P in phase with the optical scanning position can be immediately obtained. Further, even for a phase shift different for each line, it is possible to perform a correction corresponding to each shift amount, and furthermore, it is possible to realize with a simple configuration.

Although the setting of the reference voltage in the reference voltage register 19 is performed by the CPU 14, a configuration in which the reference voltage is obtained by calculation from the drive signal d of the memory 16 is also possible. In that case, the calculation is performed in consideration of the values of the zoom function and the offset function, and the output of the voltage comparison circuit 18 is obtained.

Further, in the above-described second embodiment, two-dimensional scanning has been described as an example. However, the present invention is not limited to this. In addition, regardless of raster scanning or reciprocating scanning, the optical scanning position and the sampling clock of the sampling clock are determined. This enables phase correction.

(3) Next, a third embodiment of the present invention will be described with reference to the drawings. 4 and 5 described above.
The same parts as those described above are denoted by the same reference numerals, and detailed description thereof will be omitted.

FIG. 8 is a configuration diagram of the scanning laser microscope, and FIG. 9 is a specific configuration diagram of the phase correction circuit 30.

The memory 30 stores only the drive signal d and the trigger pattern k for the X-direction drive circuit 12 and the Y-direction drive circuit 13, and the sampling clock p used for image processing is stored in the phase correction circuit 31. .

As shown in FIG. 8, the phase correction circuit 31 includes a reference voltage register 19, a voltage comparison circuit 32, a memory 33, a memory read control circuit 34 for performing read control on the memory 33, and a phase difference detection circuit. 35 are provided.

The sampling clock p is previously stored in the memory 33 by a CPU or the like.

The voltage comparison circuit 32 compares the electric signal output from the sensor 8 with a reference voltage, and when a predetermined condition is met, for example, the laser beam is emitted by the X-direction galvanometer mirror 10 and the Y-direction galvanometer mirror 12. It has a function of sending an effective signal to the memory readout control circuit 34 during the forward and backward passes when Q is scanned on the sample 4.

When the memory read control circuit 34 receives the valid signal from the voltage comparison circuit 32, it operates an internal memory address counter to count the CLK signal. It has a function of sequentially reading data, that is, the sampling clock p.

Next, the operation of the scanning laser microscope configured as described above will be described.

The CPU 14 writes the waveform of the drive signal d for driving the X-direction galvanometer mirror 10 and the Y-direction galvanometer mirror 11 and the trigger pattern k in the memory 30 in advance. Further, the CPU 14 writes a sampling clock p corresponding to the drive signal d in the memory 33 in the phase correction circuit 31.

When the scan is started, the start timing of the scan is output from the CPU 14 to the memory read control circuit 15. The memory read control circuit 15 counts the CLK that is a reference for memory read by an internal counter,
The result is output to the memory 30 as a memory address.

The memory 30 sequentially outputs the drive signal d and the data of the trigger pattern k of the corresponding address in accordance with the input memory address. The output drive signal d is input to the D / A converter 17 and converted into an analog signal, and then drives the X and Y direction galvanometer mirrors 10 and 11 through the X and Y direction drive circuits 12 and 13. I do. The sample 4 is irradiated with the laser light Q deflected by the X and Y direction galvanometer mirrors 10 and 11, and the reflected light or the fluorescence R is detected by the photodetector 6 and transmitted to the image processing system 7.

On the other hand, the sensor 8 detects the optical scanning position of the laser beam Q and sends out the electric signal to the phase correction circuit 31. The phase difference detection circuit 35 of the phase correction circuit 31
Detects the phase difference by comparing with the trigger pattern k read from the memory 30. In the case of the present embodiment,
Since the detected phase difference is not used in the subsequent processing, the output of the phase difference detection circuit 35 is omitted.

When the phase difference detection is required as described above, the trigger pattern k itself is not required, so that only the drive waveform needs to be stored in the memory, and the configuration is further simplified.

The output signal of the sensor 8 gives a timing to start reading from the memory 35 in the phase correction circuit 31 and generates a sampling clock p.

Here, the read operation of the memory 35 in the phase correction circuit 31 will be described in detail.

When the output signal of the sensor 8 is input,
The voltage comparison circuit 32 compares the value with a preset reference voltage value of the reference voltage register 19, outputs a valid signal when the condition is satisfied, and transmits the signal to the memory read control circuit 34.

When receiving the valid signal from voltage comparison circuit 32, memory read control circuit 34 operates the memory address counter and counts the CLK signal.

The memory 33 receives the output of the memory address counter and sequentially reads out the data at the corresponding address. That is, since the sampling clock p is read from the memory 33 in synchronization with the output signal of the sensor 8 indicating the optical scanning position of the laser light Q, this sampling clock p The position and the phase match.

As described above, in the third embodiment, the sampling clock p is stored in the memory 33 in the phase correction circuit 31, and the output signal of the sensor 8 for detecting the scanning position of the laser light Q is predetermined. When the above condition is satisfied, the validity signal is output from the voltage comparison circuit 32, the sampling clock p is sequentially read from the memory 33, and is sent to the image processing system 7.
Needless to say, the same effect as that of the embodiment can be obtained, and the sensor 8 indicating the optical scanning position can be provided even if the phase difference detecting circuit 35 is provided but the phase difference is not detected.
By simply reading out the memory 33 storing the pattern of the sampling clock p in synchronization with the output signal, the sampling clock p in phase with the optical scanning position can be obtained. This is because the waveform pattern of the drive signal d and the pattern of the sampling clock p are stored in separate memories 30 and 3.
3 and read out.

By setting the comparison conditions of the voltage comparison circuit 32, not only the sampling clock p but also signals necessary for operations and processing to be synchronized with the optical scanning position can be set at arbitrary timing with respect to the optical scanning position. Can occur in.

[0094]

As described above in detail, according to the present invention, the phase correction between the optical scanning position and the sampling clock is performed in real time for each line, and this can be realized with a simple configuration to reduce distortion and deterioration. It is possible to provide a scanning laser microscope capable of obtaining a non-image.

[Brief description of the drawings]

FIG. 1 is a configuration diagram showing a first embodiment of a scanning laser microscope according to the present invention.

FIG. 2 is a diagram showing a waveform of a sine-wave drive signal when scanning a sample with laser light.

FIG. 3 is a schematic diagram showing reciprocal scanning of laser light on a sample.

FIG. 4 is a configuration diagram showing a second embodiment of the scanning laser microscope according to the present invention.

FIG. 5 is a specific configuration diagram of a phase correction circuit in the scanning laser microscope.

FIG. 6 is a diagram showing waveforms of a sampling clock and a trigger pattern corresponding to a sine wave driving signal when scanning a sample with laser light.

FIG. 7 is a diagram showing a correction operation of a sampling clock corresponding to a sine wave drive signal when scanning with laser light.

FIG. 8 is a configuration diagram showing a scanning laser microscope according to a third embodiment of the present invention.

FIG. 9 is a specific configuration diagram of a phase correction circuit in the scanning laser microscope.

FIG. 10 is a diagram for explaining a conventional technique.

[Explanation of symbols]

1: light source, 2: deflection means, 3: drive circuit, 4: specimen, 5: storage means, 6: photodetector, 7: image processing system, 8: sensor, 9: phase correction means, 10: deflection means ( X-direction galvanometer mirror, 11: deflection means (Y-direction galvanometer mirror), 12: X-direction drive circuit, 13: Y-direction drive circuit, 14: CPU, 15: memory read control circuit, 16: memory, 17: A / D converter, 18: voltage comparison circuit, 19: reference voltage register, 20: FIFO (first in-first out)
Control circuit, 21: FIFO, 22: half mirror, 30: memory, 31: phase correction circuit, 32: voltage comparison circuit, 33: memory, 34: memory read control circuit, 35: phase difference detection circuit.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G043 AA03 EA01 FA02 HA02 KA09 LA03 NA06 2G059 AA05 EE02 EE07 FF03 GG01 JJ13 JJ15 KK04 MM10 MM14 PP04 2H045 AB54 AB62 BA13 CA72 CA97 DA31 2H052 AA07 AA09 AC

Claims (3)

[Claims] 1. A deflecting means for scanning a sample with laser light, a light detecting means for detecting light from the sample to obtain an image signal, a sampling clock and a trigger for outputting an image signal obtained by the light detecting means Image processing means for performing image processing according to a pattern; scanning position detecting means for detecting a scanning position of the laser light on the sample by the deflecting means; and a scanning position of the laser light detected by the scanning position detecting means. A scanning laser microscope comprising: a phase correction unit that matches a phase of the scanning position of the laser light with the sampling clock supplied to the image processing unit based on the sampling clock. 2. The phase correction means stores the sampling clock and the trigger pattern, detects a phase difference between a scanning position of the laser beam detected by the scanning position detection means and a trigger pattern, and detects the phase difference. 2. The scanning laser microscope according to claim 1, wherein the scanning laser microscope has a function of giving the sampling clock to the image processing means according to a phase difference. 3. The phase correction means stores the sampling clock, reads out the sampling clock so as to match the scanning position of the laser beam detected by the scanning position detection means, and gives the sampling clock to the image processing means. The scanning laser microscope according to claim 1, having a function.