JP4020633B2 - Scanning laser microscope, drive waveform data generation method, and control program therefor - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a technique for automatically generating drive waveform data for driving a scanning mechanism mounted on a scanning laser microscope based on operating characteristic data of the scanning mechanism.
[0002]
[Prior art]
Conventionally, a scanning laser that scans a light beam two-dimensionally on the sample surface, converts fluorescence, transmitted light, or reflected light from the sample into an electrical signal by photoelectric conversion, and acquires image data corresponding to the sample A microscope is known.
[0003]
As a scanning mechanism mounted on this scanning laser microscope, a galvanometer scanner is widely used. This galvanometer scanner is driven based on the drive waveform data registered (recorded) in the system (system controller), and the drive waveform data includes the frequency characteristics, current characteristics, and delay characteristics of the mounted galvanometer scanner. These are created based on the operation characteristics such as the scanning conditions and the scanning conditions such as the scanning range and the scanning speed. Therefore, when the model of the installed galvanometer scanner is changed, all the drive waveform data that has been registered in the system is discarded, and a new drive waveform is created based on the operating characteristics of the newly installed galvanometer scanner. The data had to be recreated and re-registered in the system. Therefore, every time the model of the mounted galvanometer scanner is changed, a burden (for example, a burden on the user or the like) related to the creation and registration of the drive waveform data is generated.
[0004]
Therefore, in order to eliminate such a burden, for example, in Japanese Patent Application Laid-Open No. 11-231253, reference drive waveform data is registered in advance, and the galvanometer scanner is driven using the reference drive waveform data. The deflection position data corresponding to the deflection position of the galvanometer scanner at each point is obtained as digital data via the A / D converter, that is, the deflection position of the galvanometer scanner is fed back, and the obtained deflection position of the galvanometer scanner is obtained. A technique has been proposed in which frequency analysis is performed on the data using a Fourier transform by a digital signal processor, and based on the analysis result, drive waveform data serving as a reference is optimized by the mounted galvanometer scanner.
[0005]
[Problems to be solved by the invention]
However, in this proposal, there is a problem that a recording area for preliminarily recording drive waveform data serving as a reference must be secured, and a recording capacity of a recording medium to be mounted increases. In this proposal, the frequency analysis using Fourier transform is performed on the deflection position data corresponding to the feedback deflection position, so that the control processing becomes complicated, and the digital signal processor is used to perform the Fourier transform. There is a problem that the configuration becomes complicated and the component cost increases.
[0006]
In addition, there has not been proposed a technique for simultaneously solving such a problem related to the above proposal and the above-described problem that a burden associated with creation and registration of drive waveform data every time the model of the galvanometer scanner is changed. .
SUMMARY OF THE INVENTION In view of the above circumstances, an object of the present invention is to eliminate the need for registration of changes in drive waveform data associated with a change in the scanning mechanism model, simplify the configuration, and reduce the cost of components. A method and its control program are provided.
[0007]
[Means for Solving the Problems]
According to the first aspect of the present invention, the sample is irradiated with a light beam from a light source, the sample surface is two-dimensionally scanned with the light beam by a mounted scanning mechanism, and at least fluorescence, reflected light, or transmission from the sample. A drive waveform data generation method for the scanning mechanism in a scanning laser microscope that converts light into an electrical signal by photoelectric conversion to acquire image data, the detection step detecting the model of the scanning mechanism; Corresponds to the model detected in the detection step Of the scanning mechanism Regarding operating characteristics Acquired in a characteristic data acquisition step for acquiring characteristic data, a scanning condition acquisition step for acquiring a scanning condition including at least a scanning range or a scanning speed when two-dimensionally scanning the sample surface, and the characteristic data acquisition step A generation step of generating drive waveform data of the scanning mechanism based on characteristic data and the scanning condition acquired in the scanning condition acquisition step. is there.
[0008]
According to the above method, based on the characteristic data (for example, frequency characteristic, current characteristic, delay characteristic, etc.) of the scanning mechanism and the scanning condition intended by the user (observer, etc.), the optimum driving waveform data for the scanning mechanism. Is automatically generated, so that it is not necessary to register the change of the drive waveform data. Further, since it is not necessary to provide a recording area for recording drive waveform data in advance, the recording capacity of the recording medium can be reduced. In addition, since it is not necessary to perform complicated control processing such as feedback analysis of the deflection position of the scanning mechanism, the configuration can be simplified, and it is not necessary to mount components for the complicated control processing. Parts costs can also be reduced.
[0009]
According to a second aspect of the present invention, in the first aspect of the invention, in the characteristic data acquisition step, the scanning laser microscope is provided at least when the scanning laser microscope is started, initialized, or started. The characteristic data of the scanning mechanism recorded on the recording medium is obtained.
[0010]
According to this method, the characteristic data is acquired from the recording medium when the scanning laser microscope is started, initialized, or when scanning is started, and the computer or the like that controls the operation of the scanning laser microscope is used as necessary. It can be read out.
According to a third aspect of the present invention, in the first aspect of the present invention, in the generation step, the deflection angle of the scanning mechanism is set so that the center of the generated drive waveform data coincides with the center of the periodic signal for sample image acquisition. The drive waveform data is generated with reference to the center of the deflection range.
[0011]
According to this method, since the center of the generated drive waveform data coincides with the center of the periodic signal for sample image acquisition, an appropriate sample image can be acquired.
According to a fourth aspect of the present invention, a specimen is irradiated with a light beam from a light source, and the specimen surface is two-dimensionally scanned with the light beam by an on-board scanning mechanism, and at least fluorescence, reflected light, or transmission from the specimen. A scanning laser microscope for acquiring image data by converting light into an electrical signal by photoelectric conversion, and detecting means for detecting a model of the scanning mechanism; Corresponds to the model detected by the detection means Of the scanning mechanism Regarding operating characteristics Characteristic data acquisition means for acquiring characteristic data, scanning condition acquisition means for acquiring a scanning condition including at least a scanning range or scanning speed when two-dimensionally scanning the sample surface, and characteristics acquired by the characteristic data acquisition means And a generating unit that generates drive waveform data of the scanning mechanism based on the data and the scanning condition acquired by the scanning condition acquiring unit.
[0012]
According to the above configuration, based on the characteristic data (for example, frequency characteristic, current characteristic, delay characteristic, etc.) of the scanning mechanism and the scanning condition intended by the user (observer, etc.), the optimum driving waveform data for the scanning mechanism. Is automatically generated, so that it is not necessary to register the change of the drive waveform data. Further, since it is not necessary to provide a recording area for recording drive waveform data in advance, the recording capacity of the recording medium can be reduced. In addition, since it is not necessary to perform complicated control processing such as feedback analysis of the deflection position of the scanning mechanism, the configuration can be simplified, and it is not necessary to mount components for the complicated control processing. Parts costs can also be reduced.
[0013]
According to a fifth aspect of the present invention, a specimen is irradiated with a light beam from a light source, and the specimen surface is two-dimensionally scanned with the light beam by an on-board scanning mechanism. A control program for causing a computer to perform drive waveform data generation control of the scanning mechanism in a scanning laser microscope that obtains image data by converting light into an electrical signal by photoelectric conversion, and detects a model of the scanning mechanism A detection step; Corresponds to the model detected by the detection means Of the scanning mechanism Regarding operating characteristics Acquired in a characteristic data acquisition step for acquiring characteristic data, a scanning condition acquisition step for acquiring a scanning condition including at least a scanning range or a scanning speed when two-dimensionally scanning the sample surface, and the characteristic data acquisition step A control program that causes the computer to execute a generation step of generating drive waveform data of the scanning mechanism based on characteristic data and the scanning condition acquired in the scanning condition acquisition step.
[0014]
By causing the computer to execute the above control program, the scanning mechanism can be controlled based on the characteristic data (for example, frequency characteristics, current characteristics, delay characteristics, etc.) of the scanning mechanism and the scanning conditions intended by the user (observer). Since optimum drive waveform data is automatically generated, change registration of drive waveform data becomes unnecessary. Further, since it is not necessary to provide a recording area for recording drive waveform data in advance, the recording capacity of the recording medium can be reduced. In addition, since it is not necessary to perform complicated control processing such as feedback analysis of the deflection position of the scanning mechanism, the configuration can be simplified, and it is not necessary to mount components for the complicated control processing. Parts costs can also be reduced.
[0015]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below with reference to the drawings.
FIG. 1 (a) is a configuration example of a scanning laser microscope according to the first embodiment of the present invention, and FIG. 1 (b) is a diagram showing a data structure example of a recording medium included in the configuration. is there.
[0016]
In FIG. 2A, the scanning laser microscope according to the present embodiment includes a light source 1, a scanning mechanism 2, an objective lens 3, a stage 4, a photodetector 5, an A / D converter 6, and a D / A converter 7. , A PC (Personal Computer) 8, a frame memory 9, a display device 10 and the like.
[0017]
The light source 1 emits a light beam.
The scanning mechanism 2 is a confocal scanner such as a galvanometer scanner, for example, and in accordance with the drive waveform data (scanner drive waveform data) output from the D / A converter 7, the light beam emitted from the light source 1 in the XY direction. Shake in two dimensions.
[0018]
The objective lens 3 condenses the light beam shaken by the scanning mechanism 2.
On the stage 4, a sample (specimen) is placed.
The photodetector 5 detects transmitted light (or reflected light, fluorescence, etc.) from the sample and converts the detected light into an electrical signal (analog data).
[0019]
The A / D converter 6 converts the analog data detected by the photodetector 5 according to the sample into digital data.
The D / A converter 7 converts the scanner drive waveform data (analog data) of the scanning mechanism 2 generated by the PC 8 into digital data.
[0020]
The PC 8 includes a CPU (Central Processing Unit) 81, a recording medium 82, a scanning mechanism acquisition unit 83, a characteristic data acquisition unit 84, a scanning condition acquisition unit 85, and the like, and controls the operation of the entire scanning laser microscope. To do.
As shown in FIG. 5B, the recording medium 82 stores control programs such as a control program 821 and a scanner drive waveform generation program 822, and also frequency characteristics, current characteristics, and delay characteristics of the scanning mechanism 2. A characteristic data file 823 and the like relating to operation characteristics such as the above are recorded. As the recording medium 82, for example, a floppy (registered trademark) disk, a hard disk, a CD-ROM, an MO (magneto-optic), or the like is applicable. Each program recorded in the recording medium 82 may be downloaded from another server computer or the like via a network (not shown) and executed.
[0021]
The CPU 81 controls the operation of the scanning laser microscope according to the control program 821 recorded on the recording medium 82 and drives the scanner of the scanning mechanism 2 according to the scanner drive waveform generation program 822 recorded on the recording medium 82. Performs processing to automatically generate waveform data.
[0022]
The scanning mechanism acquisition unit 83 detects (acquires) the model of the scanning mechanism 2 mounted on the scanning laser microscope under the control of the CPU 81, and outputs the detection result (model data corresponding to the detected model). The CPU 81 is notified. For the detection of the model, for example, a model correspondence table (definition file) or the like is recorded in the recording unit 82 in advance, and the correspondence table is referred to as necessary, while the scanning mechanism 2 is mounted. Based on the presence or absence of electrical connection, identification data sent from the scanning mechanism 2, etc., the corresponding model of the scanning mechanism 2 is detected.
[0023]
The characteristic data acquisition means 84 acquires characteristic data from the characteristic data file 823 recorded on the recording medium 82 under the control of the CPU 81 and notifies the CPU 81 of the acquired characteristic data.
Based on the control of the CPU 81, the scanning condition acquisition unit 85 acquires a scanning condition such as a scanning range and a scanning speed, which is an instruction from a user (observer or the like), and scan condition data corresponding to the acquired scanning condition. The CPU 81 is notified. The instruction from the user is performed via, for example, a GUI (Graphical User Interface).
[0024]
The frame memory 9 stores digital data (image data) corresponding to the sample output from the CPU 81.
The display device 10 displays a specimen image or the like corresponding to the digital data stored in the frame memory 9.
[0025]
With such a configuration, the light beam emitted from the light source 1 is shaken in the two-dimensional direction (planar direction) by the scanning mechanism 2 in accordance with the generated scanner drive waveform data, is condensed by the objective lens 3, and the stage 4 The upper sample surface is scanned. The transmitted light (or reflected light, fluorescence, etc.) from the sample surface is converted into an electric signal by the photodetector 5, converted into digital data by the A / D converter 6, and input to the CPU 81, and the frame memory 9 is stored. The specimen image corresponding to the digital data is displayed on the display device 10.
[0026]
Next, control processing performed by the CPU 81 of the PC 8 in the scanning laser microscope having the above-described configuration will be described. This control process is realized by the CPU 81 reading and executing the control program 821 and the scanner drive waveform generation program 822 recorded on the recording medium 82.
[0027]
FIG. 2 is a flowchart showing the processing contents of the scanning process at the start of scanning by the scanning mechanism 2 according to the first embodiment of the present invention.
In the figure, first, in S201, model data corresponding to the model of the mounted scanning mechanism 2 is acquired via the scanning mechanism acquisition unit 82 described above.
[0028]
In S202, it is determined whether or not the characteristic data file corresponding to the model data acquired in the previous step is recorded in the recording means 82. This determination is made according to whether or not the characteristic data acquisition unit 84 can acquire the characteristic data file corresponding to the model data acquired in the previous step. In this determination, if the determination result is YES, the process proceeds to S203, and if the determination result is NO, the process proceeds to S208.
[0029]
In S 203, characteristic data is acquired from the corresponding characteristic data file 823 via the characteristic data acquisition unit 84, and this characteristic data is stored in the internal register of the CPU 81.
In S <b> 204, scan range data corresponding to the scan range in the XY direction (two-dimensional direction) is acquired via the scan condition acquisition unit 85 described above, and the scan range data is stored in an internal register of the CPU 81. For example, data corresponding to the scanning range is stored such that the scanning range in the X direction (horizontal direction) is 512 pixels, the scanning range in the Y direction (vertical direction) is 512 pixels, and the like.
[0030]
In S205, the scanning speed data corresponding to the scanning speed is acquired via the above-described scanning condition acquisition unit 85, and the scanning speed data is stored in the internal register of the CPU 81.
In S206, characteristic data corresponding to the scanning mechanism 2 stored in the internal register of the CPU 81 in S203 described above, scanning range data stored in the internal register of the CPU 81 in S204, and stored in the internal register of the CPU 81 in S205. Based on the scanning speed data, the scanner driving waveform data optimized for the mounted scanning mechanism 2, that is, the X direction scanner driving waveform data for swinging the light beam in the X direction (horizontal direction), and the light beam. Scanner drive waveform generation processing for generating Y-direction scanner drive waveform data for oscillating in the Y direction (vertical direction) is performed. The scanner drive waveform generation process will be described later.
[0031]
In S207, the X-direction and Y-direction scanner drive waveform data (digital data) generated in the previous step is output to the D / A converter 7, converted into analog data by the D / A converter 7, and scanned. The scanning mechanism 2 is driven in accordance with the scanner driving waveform data output to the mechanism 2 and converted into the analog data. Thereafter, this flow ends.
[0032]
In S208, error processing is performed. In this error processing, for example, a display for notifying the user that the characteristic data file corresponding to the mounted scanning mechanism 2 is not recorded on the recording medium 82 is displayed on the display device 10. Thereafter, this flow ends.
[0033]
Next, the scanner drive waveform generation process, which is the process of S206 described above, will be described with reference to FIGS.
FIG. 3 is a flowchart showing the processing contents of the scanner drive waveform generation processing, and FIG. 4 is an example of the generated X direction (or Y direction) scanner drive waveform data. In FIG. 4, the horizontal axis represents time, and the vertical axis represents the deflection angle of the scanning mechanism 2.
[0034]
In FIG. 3, first, in S301, the target of the generated scanner drive waveform data is set in the X-axis direction (X direction, horizontal direction) (Axis = X).
In S302, the scanning range (ScanSize) in the direction (in this case, the X-axis direction) of the scanner drive waveform data generated from the scanning range data stored in the internal register of the CPU 81 in S204 of FIG. To be acquired.
[0035]
In S303, the shake is performed from the characteristic data file 823 corresponding to the scanning mechanism 2 stored in the internal register of the CPU 81 in S203 of FIG. 2 described above in the target direction of the scanning mechanism 2 (in this case, the X-axis direction). The maximum deflection angle that can be read is read.
[0036]
In S304, a counter value for counting the number of scanner drive waveform data in the section T1 to be sampled shown in FIG. 4 is initialized (ActiveDataNum = 0).
In S305, the inclination of the scanner drive waveform data is obtained. That is, the increment of the deflection angle, which is the inclination in the target direction (in this case, the X-axis direction) in the section T1 to be sampled, is obtained by the following equation (1).
[0037]
Increment of deflection angle = maximum deflection angle / (total number of scanner drive waveform data in sampling period T1 × sampling period) Equation (1)
In Equation (1), the maximum deflection angle is the maximum deflection angle that can be shaken in the target direction (in this case, the X-axis direction) obtained in S303 described above. Further, the total number of scanner drive waveform data in the section T1 to be sampled becomes 512 when the target direction is the X-axis direction when the scanning range is the aforementioned 512 × 512 pixels, for example. The sampling period is based on the scanning speed data stored in the internal register of the CPU 81 in S205 of FIG.
[0038]
In S306, scanner drive waveform data in the grace period T2 is generated from the point P shown in FIG. 4 where the deflection angle in the direction (in this case, the X-axis direction) targeted by the scanning mechanism 2 is minimized. The grace period is an interval for securing a time required for the scanning mechanism 2 to follow the input signal. In this step, delay characteristic data is read from the characteristic data stored in the internal register of the CPU 81 in S203 of FIG. 2 described above, and the number of scanner drive waveform data necessary for the grace period T2 is determined based on the delay characteristic data. Based on the calculated number of scanner drive waveform data, the scanner drive waveform data in the grace period T2 is generated.
[0039]
In S307 and S308, scanner drive waveform data in the section T1 to be sampled after the grace period T2 is additionally generated one by one in order from the grace period T2 side.
That is, in S307, one scanner drive waveform data is additionally generated based on the deflection angle increment calculated in S305, and the counter value is incremented (ActiveDataNum + = 1).
[0040]
In S308, it is determined whether or not the counter value is equal to the scanning range in the target direction (in this case, the X-axis direction), that is, the total number of scanner drive waveform data in the section T1 to be sampled (ActiveDataNum = ScanSize). ?), If this determination result is YES, the process proceeds to 309, and if it is NO, the process returns to S307. In this way, the processing of S307 and S308 is repeated until ActiveDataNum = ScanSize, and additional scanner drive waveform data in the section T1 to be sampled in the target direction (in this case, the X-axis direction) is generated. .
[0041]
In S309, it is determined whether or not the target of the generated scanner drive waveform data is in the Y-axis direction (Y direction, vertical direction). If this determination result is YES, the process proceeds to S311, and if NO The process proceeds to 310.
In S310, the target of the generated scanner drive waveform data is set in the Y-axis direction (Axis = Y), and the process returns to S302. As a result, the target is set in the Y-axis direction, and the above-described processing is repeated, and scanner drive waveform data in the Y-axis direction is generated in the grace period T2 and the sampling target period T1.
[0042]
In S311, X-axis direction and Y-axis direction scanner drive waveform data are additionally generated in the grace period T2 ′ after the sampling period T1 shown in FIG. That is, in this step, the delay characteristic data is read from the characteristic data stored in the internal register of the CPU 81 in S203 of FIG. 2 described above, and the scanner drive waveform necessary for the grace period T2 ′ based on the delay characteristic data. The number of data is calculated, and the scanner drive waveform data in the grace period T2 ′ is additionally generated based on the calculated number of scanner drive waveform data.
[0043]
In S312, scanner drive waveform data is generated in the blanking intervals T3 and T3 ′ shown in FIG. 4, which are additionally generated in the X-axis direction and Y-axis direction scanner drive waveform data generated up to the previous step. . The blanking interval means that the scanning mechanism 2 completes one scan, that is, one line scan in the X-axis direction, or one frame scan in the Y-axis direction, and reaches the maximum deflection angle. This is a section for returning to the original deflection angle (minimum deflection angle etc.) after reaching. In this step, current characteristic data is read from the characteristic data stored in the internal register of the CPU 81 in S203 of FIG. 2 described above, and based on the current characteristic data, as scanner drive waveform data in the blanking intervals T3 and T3 ′. Further, sine waveform data having a quarter period is returned to return the scanning mechanism 2 to the original deflection angle at the maximum frequency. Thereafter, this flow ends.
[0044]
The above processing is the scanning processing at the start of scanning by the scanning mechanism 2 according to the first embodiment, and this processing is performed by the CPU 81 of the PC 8 so that the scanning mechanism 2 mounted at the time of scanning starts. Based on the corresponding characteristic data file 823 and the scanning condition instructed by the user, the scanner driving waveform data optimized for the scanning mechanism 2 in the X-axis direction and the Y-axis direction is automatically generated. Accordingly, the scanning mechanism 2 is driven.
[0045]
As described above, according to the first embodiment, the scanner drive waveform data optimized for the mounted scanning mechanism 2 is automatically generated based on the characteristic data file 823 and the scanning conditions. Therefore, it is not necessary to previously record the scanner drive waveform data on the recording medium included in the scanning laser microscope, and the recording capacity of the recording medium can be reduced.
[0046]
Further, when a scanning mechanism of a different model is mounted, the contents of the characteristic data file 823 recorded on the recording medium 82 are simply rewritten with the contents of the characteristic data file corresponding to the newly mounted scanning mechanism 2. Thus, since the scanner driving waveform data optimized for the newly mounted scanning mechanism 2 is automatically generated, the scanner driving waveform based on the characteristic data of the newly mounted scanning mechanism as in the prior art. The burden of creating data and registering it can be eliminated.
[0047]
In addition, the control process described above is performed by the CPU 81 of the PC 8 that controls the operation of the entire scanning laser microscope, whereby the configuration of the scanning laser microscope itself can be simplified. Such costs can be reduced.
[0048]
Next, a second embodiment of the present invention will be described.
This embodiment is different from the first embodiment described above in scanner drive waveform generation processing. Therefore, here, the scanner drive waveform generation processing will be described.
FIG. 5 is a flowchart showing the processing contents of the scanner drive waveform generation processing according to the second embodiment of the present invention. The scanner drive waveform generation processing according to the present embodiment is also realized by the CPU 81 of the PC 8 reading and executing the control program 821 and the scanner drive waveform generation program 822 recorded in the recording unit 82.
[0049]
In the figure, first, in S501 through S505, the same processing as the processing in S301 through S305 shown in FIG. 3 is performed.
In the subsequent processes of S506 to S509, scanner drive waveform data in the section T1 to be sampled is generated. First, in the processing of S506 and S507, scanner drive waveform data in a half section on the maximum deflection angle side of the section T1 to be sampled is sequentially generated one by one toward the maximum deflection angle side, and the subsequent S508 and S509 are performed. In the processing, scanner drive waveform data in a half section on the minimum deflection angle side of the section T1 to be sampled is generated one by one in order toward the minimum deflection angle side.
[0050]
That is, first, in S506, one scanner drive waveform data is generated in the direction in which the deflection angle increases with reference to the center of the deflection range of the deflection angle in the target direction (in this case, the X-axis direction), and the counter The value is incremented (ActiveDataNum + = 1).
[0051]
In S507, the number of scanner drive waveform data generated from the center of the deflection range of the deflection angle in the target direction (in this case, the X-axis direction) toward the direction in which the deflection angle increases is acquired in S502 described above. It is determined whether or not the scan range is half (ActiveDataNum = ScanSize / 2?). If the determination result is YES, the counter is initialized (ActiveDataNum = 0), and the process proceeds to S509. If NO, The process returns to S506, and the above-described process is repeated. In this manner, the processing of S506 and S507 is repeated until ActiveDataNum = ScanSize / 2, and scanner drive waveform data in the half section on the maximum deflection angle side of the section T1 to be sampled is generated.
[0052]
In S508, one scanner drive waveform data is generated in the direction in which the deflection angle decreases with reference to the center of the deflection range of the deflection angle in the target direction (X-axis direction in this case), and the counter value is incremented. (ActiveDataNum + = 1).
In S509, the number of scanner drive waveform data generated in the direction in which the deflection angle decreases from the center of the deflection range of the deflection angle in the target direction (X-axis direction in this case) is the scan acquired in S502 described above. It is determined whether or not it is half of the range (ActiveDataNum = ScanSize / 2?). If this determination result is YES, the process proceeds to S510, and if it is NO, the process returns to S508, and the above-described process is performed. Repeated. In this manner, the processing of S508 and S509 is repeated until ActiveDataNum = ScanSize / 2, and scanner drive waveform data in the half section on the minimum deflection angle side of the section T1 to be sampled is generated.
[0053]
In S510 and S511, the same processing as S309 and S310 shown in FIG. 3 is performed.
In S512, the scanner drive waveform data in the grace period T2 and T2 ′ is added to the X-axis direction and Y-axis direction scanner drive waveform data generated so far (scanner drive waveform data in the section T1 to be sampled), respectively. Generated. That is, the delay characteristic data is read from the characteristic data file 823 stored in the internal register of the CPU 81 in S203 of FIG. 2 described above, and based on the delay characteristic data, the scanner drive waveform necessary for the grace periods T2 and T2 ′. The number of data is calculated, and based on the calculated number of scanner drive waveform data, the scanner drive waveform data in the grace period T2 and T2 ′ is additionally generated on the extension line of the section T1 to be sampled.
[0054]
In S513, the same process as the process of S312 shown in FIG. 3 is performed. Thereafter, this flow ends.
The above processing is the scanner driving waveform generation processing according to the second embodiment. When this processing is performed by the CPU 81 of the PC 8, the characteristic data file 823 corresponding to the mounted scanning mechanism 2 and the instruction from the user are provided. Based on the scanned conditions, the X-axis and Y-axis scanner drive waveform data optimized for the scanning mechanism 2 is generated based on the center of the deflection range of the deflection angle of the scanning mechanism 2 Is done.
[0055]
As described above, according to the second embodiment, the scanner drive waveform data in the X-axis direction and the Y-axis direction is generated with reference to the center of the deflection range of the deflection angle. It is possible to match the center of the periodic signal for sample image acquisition corresponding to one scan in the X-axis direction (one line scan) or one scan in the Y-axis direction (one frame scan). Can be acquired.
[0056]
Next, a third embodiment of the present invention will be described.
This embodiment is different from the first embodiment described above in that a plurality of characteristic data files corresponding to a plurality of types of scanning mechanisms 2 are recorded in the recording unit 82 in advance, and scanning is performed accordingly. The scanning process at the start of scanning by the mechanism 2 is different. Therefore, here, scanning processing at the start of scanning by the scanning mechanism 2 will be described.
[0057]
FIG. 6 is a flowchart showing the processing contents of the scanning processing at the start of scanning by the scanning mechanism 2 according to the third embodiment of the present invention. Note that the scanning process at the start of scanning by the scanning mechanism 2 according to the present embodiment is also performed by the CPU 81 of the PC 8 reading and executing the control program 821 and the scanner drive waveform generation program 822 recorded in the recording unit 82. Realized.
[0058]
In this figure, in S601, model data corresponding to the model of the mounted scanning mechanism 2 is acquired in the same manner as in the process of S201 shown in FIG.
In S602, the model data acquired in the previous step is associated with the corresponding characteristic data file among a plurality of characteristic data files recorded in the recording unit 82 in advance.
[0059]
In S603, it is determined whether or not the characteristic data file corresponding to the model data acquired in S601 is recorded in the recording unit 82. This determination is made according to whether or not the model data acquired in the above-described S601 is associated with the corresponding characteristic data file in the previous step. In this determination, if the determination result is YES, the process proceeds to S604, and if the determination result is NO, the process proceeds to S609.
[0060]
In S604 to S609, processing similar to the processing of S203 to S208 shown in FIG. 2 described above is performed, and this flow ends.
The above processing is scanning processing at the start of scanning by the scanning mechanism 2 according to the third embodiment, and this processing is performed by the CPU 81 of the PC 8 so that it is recorded in the recording means 82 in advance at the start of scanning. A characteristic data file corresponding to the mounted scanning mechanism 2 is read out from the plurality of types of characteristic data files, and based on the read characteristic data file and the scanning conditions instructed by the user, Scanner drive waveform data in the X-axis direction and Y-axis direction optimized for the scanning mechanism 2 is generated, and the scanning mechanism 2 is driven according to the scanner drive waveform data.
[0061]
As described above, according to the third embodiment, since a plurality of characteristic data files corresponding to a plurality of types of scanning mechanisms are recorded on the recording medium 82 in advance, when the recording medium 82 is replaced with a scanning mechanism of a different model, There is no need to rewrite the characteristic data file, and optimized scanner drive waveform data can be generated more efficiently.
[0062]
In the first to third embodiments, the scanner drive waveform generation process is performed at the start of scanning by the scanning mechanism 2, but may be performed before the start of scanning.
In the first to third embodiments, when the scanning mechanism 2 starts scanning, a corresponding characteristic data file is acquired via the characteristic data acquisition unit 84, and the characteristic data file is stored in the internal register of the CPU 81. Although stored, this may be performed when the scanning laser microscope is activated or initialized.
[0063]
In the first to third embodiments, the generated scanner drive waveform data is composed of a sine wave and a sawtooth wave as shown in FIG. It may be.
In the first to third embodiments, the process performed by the CPU 81 may be performed by a CPU such as a workstation.
[0064]
In the first to third embodiments, the CPU 81 may directly perform the processing performed by the scanning mechanism acquisition unit 83, the characteristic data acquisition unit 84, and the scanning condition acquisition unit 85.
In the first to third embodiments, a new A / D converter is provided between the scanning mechanism 2 and the CPU 81, and the actual mechanism of the scanning mechanism 2 is provided via the A / D converter. Acquires data corresponding to the response of the deflection angle, that is, feeds back the response of the actual deflection angle, compares the acquired data corresponding to the response of the actual deflection angle with the generated scanner drive waveform data, and is mounted A difference in operating characteristics due to machine differences (individual differences) of the scanning mechanism 2 may be detected. By configuring in this way, it becomes possible to correct the contents of the characteristic data file 823 based on the difference in the detected operating characteristics, and taking into account the machine difference of the mounted scanning mechanism 2, more It is possible to generate optimized scanner drive waveform data.
[0065]
In the first to third embodiments, the control process performed by the CPU 81 of the PC 8 of the scanning laser microscope can be executed by a computer as shown in FIG. 7, for example. In this case, the control program (control program 821, scanner drive waveform generation program 822, etc.) recorded on the recording medium 82 of the PC 8 of the scanning laser microscope is a CD-ROM, floppy (registered) as shown in FIG. Trademark) disk (or MO, DVD, CD-R, CD-RW, removable hard disk, etc.) or the like is recorded on a portable recording medium 21, and the portable recording medium 21 is stored in the medium of the computer 22. The program read by the drive device 23 may be stored in a memory (RAM or hard disk or the like) 24 inside the computer 22 and the computer 22 may execute the program. Alternatively, the program is recorded in a recording means (database, etc.) 25 in an external device (server, etc.) of the information provider, transferred to the computer 22 by communication, and recorded in the internal memory 24, and the program is stored. It may be executed by the computer 22. Note that the program recorded in these programs may execute only a part of the control process performed by the CPU 81 of the PC 8 of the above-described scanning laser microscope.
[0066]
As described above, the scanning laser microscope, the drive waveform data generation method, and the control program thereof according to the present invention have been described in detail. However, the present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the spirit of the present invention. Of course, improvements and changes may be made.
<Appendix>
(Supplementary Note 1) In the generation step,
A sine wave, sawtooth wave, or other waveform optimized for the scanning mechanism based on the characteristic data acquired in the characteristic data acquisition step and the scanning condition acquired in the scanning condition acquisition step Generate drive waveform data by
A drive waveform data generation method for a scanning mechanism in a scanning laser microscope.
(Supplementary Note 2) Generation of the drive waveform data of the scanning mechanism performed in the generation step is performed by a computer or a workstation CPU that controls the operation of the scanning laser microscope.
A drive waveform data generation method for a scanning mechanism in a scanning laser microscope.
[0067]
【The invention's effect】
As described above in detail, according to the present invention, optimum drive waveform data for the scanning mechanism is automatically generated based on the characteristic data relating to the operating characteristics of the scanning mechanism and the scanning conditions intended by the user. Therefore, the change registration of the drive waveform data becomes unnecessary. Further, since it is not necessary to provide a recording area for recording drive waveform data in advance, it is possible to reduce the recording capacity of the mounted recording medium. Further, since it is not necessary to perform complicated control processing, the configuration can be simplified, and it is not necessary to mount components for the complicated control processing, so that the cost of components can be reduced.
[Brief description of the drawings]
FIG. 1A is a diagram illustrating a configuration example of a scanning laser microscope according to the first embodiment of the present invention, and FIG. 1B is a diagram illustrating a data structure example of a recording medium included in the configuration.
FIG. 2 is a flowchart showing the processing contents of a scanning process at the start of scanning by the scanning mechanism according to the first embodiment of the present invention.
FIG. 3 is a flowchart showing the processing content of scanner drive waveform generation processing.
FIG. 4 is an example of scanner drive waveform data in the X direction (or Y direction).
FIG. 5 is a flowchart showing the processing contents of scanner drive waveform generation processing according to the second embodiment of the present invention.
FIG. 6 is a flowchart showing the processing contents of a scanning process at the start of scanning by the scanning mechanism according to the third embodiment of the present invention.
FIG. 7 is a diagram illustrating an example of a recording medium on which a control program is recorded.
[Explanation of symbols]
1 Light source
2 Scanning mechanism
3 Objective lens
4 stages
5 photodetectors
6 A / D converter converter
7 D / A converter Converter
8 PC
9 frame memory
10 Display device
21 Portable storage media
22 Computer
23 Medium drive device
24 memory
25 Memory means
81 CPU
82 Recording means
83 Scanning mechanism acquisition means
84 Characteristic data acquisition means
85 Scanning condition acquisition means
821 Control program
822 Scanner drive control program
823 characteristic data file

Claims (5)

A specimen is irradiated with a light beam from a light source, and the specimen surface is two-dimensionally scanned with the light beam by an on-board scanning mechanism, and at least fluorescence, reflected light, or transmitted light from the specimen is converted into an electrical signal by photoelectric conversion. A drive waveform data generation method for the scanning mechanism in a scanning laser microscope that obtains image data by conversion,
A detection step of detecting a model of the scanning mechanism;
A characteristic data acquisition step for acquiring characteristic data relating to the operating characteristics of the scanning mechanism corresponding to the model detected in the detection step ;
A scanning condition acquisition step of acquiring a scanning condition including at least a scanning range or a scanning speed when two-dimensionally scanning the sample surface;
A generation step of generating drive waveform data of the scanning mechanism based on the characteristic data acquired in the characteristic data acquisition step and the scanning condition acquired in the scanning condition acquisition step;
A drive waveform data generation method for a scanning mechanism in a scanning laser microscope. In the characteristic data acquisition step,
Acquire characteristic data of the scanning mechanism recorded on a recording medium provided in the scanning laser microscope at least at the time of startup, initialization, or scanning of the scanning laser microscope.
2. A driving waveform data generation method for a scanning mechanism in a scanning laser microscope according to claim 1. In the generating step,
The drive waveform data is generated with reference to the center of the deflection range of the deflection angle of the scanning mechanism so that the center of the generated drive waveform data coincides with the center of the periodic signal for sample image acquisition.
2. A driving waveform data generation method for a scanning mechanism in a scanning laser microscope according to claim 1. A specimen is irradiated with a light beam from a light source, and the specimen surface is two-dimensionally scanned with the light beam by an on-board scanning mechanism, and at least fluorescence, reflected light, or transmitted light from the specimen is converted into an electrical signal by photoelectric conversion. A scanning laser microscope that converts and acquires image data,
Detecting means for detecting a model of the scanning mechanism;
Characteristic data acquisition means for acquiring characteristic data relating to operating characteristics of the scanning mechanism corresponding to the model detected by the detection means ;
Scanning condition acquisition means for acquiring a scanning condition including at least a scanning range or a scanning speed when two-dimensionally scanning the sample surface;
Generating means for generating drive waveform data of the scanning mechanism based on the characteristic data acquired by the characteristic data acquisition means and the scanning conditions acquired by the scanning condition acquisition means;
A scanning laser microscope comprising: A specimen is irradiated with a light beam from a light source, and the specimen surface is two-dimensionally scanned with the light beam by an on-board scanning mechanism, and at least fluorescence, reflected light, or transmitted light from the specimen is converted into an electrical signal by photoelectric conversion. A control program for causing a computer to execute generation control of drive waveform data of the scanning mechanism in a scanning laser microscope that converts and acquires image data,
A detection step of detecting a model of the scanning mechanism;
A characteristic data acquisition step for acquiring characteristic data relating to the operating characteristics of the scanning mechanism corresponding to the model detected in the detection step ;
A scanning condition acquisition step of acquiring a scanning condition including at least a scanning range or a scanning speed when two-dimensionally scanning the sample surface;
A generation step of generating drive waveform data of the scanning mechanism based on the characteristic data acquired in the characteristic data acquisition step and the scanning condition acquired in the scanning condition acquisition step;
A control program for causing the computer to execute.

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