Abstract:
To provide a laser scanning apparatus and a laser scanning microscope capable of securely conducting a condition setting at the time of laser scanning while suppressing a damage on a plane to be irradiated. Accordingly, a laser scanning apparatus includes a light deflecting unit disposed in a light path of laser light directed toward a plane to be scanned, user interfaces through which operational contents of the light deflecting unit are designated by a user, generating units generating driving signals of the light deflecting unit in accordance with the designated operational contents, and testing units test-driving the light deflecting unit with the driving signals while keeping the laser light off and measuring the operational contents of the light deflecting unit during the driving.

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application is a Continuation Application of International Application No. PCT/JP2007/000656, filed Jun. 20, 2007, designating the U.S., in which the International Application claims a priority date of Jul. 18, 2006, based on prior filed Japanese Patent Application No. 2006-195241, the entire contents of which are incorporated herein by reference. 
    
    
     BACKGROUND 
     1. Field 
     The present invention relates to a laser scanning apparatus and a laser scanning microscope. 
     2. Description of the Related Art 
     A general laser scanning microscope is provided with a galvanometer mirror which scans in an X direction with laser light over a sample plane and a galvanometer mirror which scans in a Y direction with laser light over the sample plane. If these two galvanometer mirrors are cooperatively controlled, it is also possible to conduct an observation (hereinafter, refer to as “free line observation”) in which a scanning trajectory of laser light (hereinafter, refer to as “scanning line”) is expressed by a free-form curve. For instance, if the free line observation is conducted with the scanning line so as to trace a cord-shaped axial filament of a nerve cell, it is also possible to capture a high-speed change generated in the axial filament. 
     However, details of an actual scanning line are not always set as designated by a user. This is because a movement of the galvanometer mirror is dependent not only on a waveform of a driving signal given from an exterior but also on an inertia of the mirror, a steepness of the scanning line, a scanning speed and the like. For this reason, in order to find an optimal scanning condition, the user needs to repeatedly conduct a trial and error process while changing the scanning conditions. 
     Meanwhile, when a sample is an organism, it is vulnerable to damage, and when the sample is fluorescent-dyed, a color fading occurs, so that a number of irradiations of laser light onto the sample has to be kept to the minimum. 
     SUMMARY 
     Accordingly, a proposition of the present invention is to provide a laser scanning apparatus and a laser scanning microscope capable of securely conducting a condition setting at the time of laser scanning while suppressing a damage on a plane to be irradiated. 
     A laser scanning apparatus of the present invention includes a light deflecting unit disposed in a light path of laser light directed toward a plane to be scanned, user interfaces through which operational contents of the light deflecting unit are designated by a user, generating units generating driving signals of the light deflecting unit in accordance with the designated operational contents, and testing units test-driving the light deflecting unit with the driving signals while keeping the laser light off and measuring the operational contents of the light deflecting unit during the driving. 
     Further, a laser scanning apparatus of the present invention includes a light deflecting unit disposed in a light path of laser light directed toward a plane to be scanned, user interfaces through which operational contents of the light deflecting unit are designated by a user, generating units generating driving signals of the light deflecting unit in accordance with the designated operational contents, and testing units test-driving the light deflecting unit with the driving signals in a state where an intensity of the laser light is lower than that used when conducting a real scanning over the plane to be scanned and measuring the operational contents of the light deflecting unit during the driving. 
     Note that the user interfaces preferably notify the user of the measured operational contents. 
     Further, the laser scanning apparatus of the present invention preferably further includes a correcting unit comparing the measured operational contents with the designated operational contents and correcting the driving signals so that the former operational contents come close to the latter ones. 
     Further, a laser scanning microscope of the present invention includes the laser scanning apparatus of the present invention and a detector detecting an intensity of light generated at the plane to be scanned. 
     According to the present invention, a laser scanning apparatus and a laser scanning microscope capable of securely conducting a condition setting at the time of laser scanning while suppressing a damage on a plane to be irradiated are realized. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is an entire structural view of the present system. 
         FIG. 2  is a structural view of a galvanometer scanner  11  and a controller  20 . 
         FIG. 3  illustrates operation flow charts of the controller  20  and a computer  21  at the time of free line observation. 
         FIG. 4  is a view showing a setting screen. 
         FIGS. 5(   a ),  5 ( b ) and  5 ( c ) are views to explain a generating method of driving waveforms. 
         FIG. 6  is a view showing a setting screen at another timing. 
         FIG. 7  is a view showing a display example of observation information. 
         FIG. 8  illustrates operation flow charts (first halves) of the controller  20  and the computer  21  in a second embodiment. 
         FIG. 9  illustrates operation flow charts (second halves) of the controller  20  and the computer  21  in the second embodiment. 
         FIG. 10  is a view showing a setting screen in the second embodiment. 
         FIG. 11  is a view showing a setting screen at another timing in the second embodiment. 
     
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENTS 
     [First Embodiment] 
     Hereinafter, a first embodiment will be explained. The present embodiment is an embodiment of a confocal fluorescence laser scanning microscope system. 
     First, an entire structure of the present system will be described. 
       FIG. 1  is an entire structural view of the present system. As shown in  FIG. 1 , the present system includes a microscope body  100 , a controller  20 , a computer  21 , a monitor  22  and an input device  23  such as a mouse and a keyboard. 
     In the microscope body  100 , a laser unit  1 , an optical fiber  7 , a collimating lens  8 , a dichroic mirror  9 , a galvanometer scanner  11 , a relay lens  14 , an objective lens  15 , a sample  16 , a collecting lens  17 , a pinhole diaphragm for confocal detection  18 , a light detector  19  and the like are disposed. The sample  16  is, for example, a fluorescent sample supported on a not-shown stage, and the galvanometer scanner  11  is provided with two galvanometer mirrors (later-described galvanometer mirrors  111 X and  111 Y) disposed in serial relationship. 
     Laser light emitted from the laser unit  1  is incident on one end of the optical fiber  7 , then propagates inside the optical fiber  7 , emitted from the other end of the optical fiber  7 , and after being turned into parallel pencil of light by the collimating lens  8 , it is incident on the dichroic mirror  9 . The laser light passes through the dichroic mirror  9 , and after being sequentially reflected by the two galvanometer mirrors of the galvanometer scanner  11 , it passes through the relay lens  14  and the objective lens  15  and is condensed to one point on the sample  16 . If the galvanometer scanner  11  is driven under this state, the laser light scans over the sample  16 . 
     A fluorescence generated at the light condensed position of the laser light on the sample  16  advances in the opposite direction along the same light path as that of the laser light, toward the dichroic mirror  9 . The fluorescence is reflected by the dichroic mirror  9 , condensed by the collecting lens  17  and passed through the pinhole diaphragm  18  to thereby remove extra light rays therefrom, and thereafter, it is incident on the light detector  19  and converted into a fluorescence signal. 
     The controller  20  of the present system synchronously controls the laser unit  1 , the galvanometer scanner  11  and the light detector  19 , to thereby repeatedly take the fluorescence signals while scanning over the sample  16  with the laser light. The fluorescence signals taken at this time are transmitted to the computer  21  as observation information, and are output to the monitor  22  or stored by the computer  21  if necessary. A user conducts an observation of the sample  16  using the observation information. 
     Next, structures of the galvanometer scanner  11  and the controller  20  will be specifically described. 
       FIG. 2  is a structural view of the galvanometer scanner  11  and the controller  20 . As shown in  FIG. 2 , the galvanometer scanner  11  is provided with two galvanometer mirrors  111 X and  111 Y. When the galvanometer mirror  111 X is driven during when the laser light is projected onto the galvanometer scanner  11 , the laser light scans over the sample  16  in a predetermined direction (X direction), and when the galvanometer mirror  111 Y is driven during when the laser light is projected onto the galvanometer scanner  11 , the laser light scans over the sample  16  in a perpendicular direction to the X direction (Y direction). 
     Between the two mirrors, the galvanometer mirror  111 X has a driver  204 X as its driving circuit coupled thereto, and the galvanometer mirror  111 Y has a driver  204 Y as its driving circuit coupled thereto. Further, the galvanometer mirror  111 X is provided with a position sensor  112 X detecting a mirror position thereof, and the galvanometer mirror  111 Y is provided with a position sensor  112 Y detecting a mirror position thereof. 
     The controller  20  includes a scanner controlling part  202  being a control circuit dedicated to the galvanometer scanner  11 , a laser controlling part  207  being a control circuit dedicated to the laser unit  1 , a detector controlling part  208  being a control circuit dedicated to the light detector  19 , a CPU  201  controlling an entire controller  20 , an interface circuit  209  performing an interface operation with the computer  21 , a ROM  201 A storing a program of the CPU  201 , and a RAM  201 B used for a temporary storage of the CPU  201 . 
     Note that what are indicated by reference numerals  205 X and  205 Y in  FIG. 2  are A/D converters which A/D convert signals output from the galvanometer scanner  11 , and what are indicated by reference numerals  203 X and  203 Y are D/A converters which D/A convert signals output from the scanner controlling part  202 . 
     Next, basic operations of the computer  21 , the controller  20  and the galvanometer scanner  11  will be described. 
     Before the observation, the computer  21  prompts the user to operate the input device  23  to get the user to input scanning conditions. The scanning conditions include at least a scanning line and a scanning speed desired by the user, and a laser intensity or the like desired by the user is normally included therein. The input scanning conditions are transmitted from the computer  21  to the controller  20 . The CPU  201  of the controller  20  recognizes the scanning conditions via the interface circuit  209 . 
     The CPU  201  records, in accordance with the recognized scanning conditions, necessary information in the laser controlling part  207 , the detector controlling part  208  and the scanner controlling part  202 , thereby setting the laser unit  1 , the light detector  19  and the galvanometer scanner  11 . 
     Incidentally, in the setting of the galvanometer scanner  11 , the CPU  201  generates a waveform of a driving signal to be given to the driver  204 X of the galvanometer scanner  11  (hereinafter, refer to as “X-driving waveform”) and a waveform of a driving signal to be given to the driver  204 Y of the galvanometer scanner  11  (hereinafter, refer to as “Y-driving waveform”) based on a set line and a set speed included in the scanning conditions, and stores information on those waveforms in a memory  202 A of the scanner controlling part  202 A. 
     Thereafter, when obtaining the observation information, the CPU  201  gives indications to the laser controlling part  207 , the detector controlling part  208  and the scanner controlling part  202  under the aforementioned set conditions, to thereby synchronously drive the laser unit  1 , the light detector  19  and the galvanometer scanner  11 . 
     At this time, the scanner controlling part  202  generates the driving signals in accordance with the information on the X-driving waveforms stored in the memory  202 A and sequentially transmits them to the driver  204 X via the D/A converter  203 X. Further, the scanner controlling part  202  generates the driving signals in accordance with the information on the Y-driving waveforms stored in the memory  202 A and sequentially transmits them to the driver  204 Y via the D/A converter  203 Y. As a result of this, the galvanometer scanner  11  is driven. 
     Further, the scanner controlling part  202  samples signals output from the position sensor  112 X (hereinafter, refer to as “X-position signals”) and signals output from the position sensor  112 Y (hereinafter, refer to as “Y-position signals”) via the A/D converters  205 X and  205 Y during the driving of the galvanometer scanner  11 , and stores them in the memory  202 A. A sampling rate is sufficiently high, and is equal to or higher than a data sampling signal frequency in the controller  20 . The X-position signals and the Y-position signals taken as above are effectively utilized at the time of free line observation to be explained next. 
     Next, operations of the controller  20  and the computer  21  at the time of free line observation will be described. 
       FIG. 3  illustrates operation flow charts of the controller  20  and the computer  21  at the time of free line observation. An operation program (control program) of the controller  20  is previously stored in the ROM  201 A of the controller  20  or the like, and an operation program (management program) of the computer  21  is previously stored in a hard disk of the computer  21  or the like. 
     (Step S 21 ) 
     First, in order to get the user to input the scanning conditions such as the scanning line, the scanning speed and the laser intensity under a GUI environment, the computer  21  displays a setting screen as shown in  FIG. 4 , for instance, on the monitor  22 . 
     As shown in  FIG. 4 , on the setting screen, an image I of an observation area of the sample  16  (within a field of view of the objective lens  15 ) is displayed. This image I is obtained by, for example, a normal observation conducted by the present system. The normal observation is for obtaining observation information by setting the laser intensity to low intensity and setting the scanning line to a stripe-shaped one. 
     Through the operation of the input device  23 , the user draws a scanning line L 1  and inputs characters indicating a scanning speed B 1 , a laser intensity B 0  and the like on the setting screen. 
     Note that on the setting screen, a testing button B 2 , a real scanning button B 3  and the like are arranged, and by selecting these buttons at a desired timing, the user can also input a testing indication and a real scanning indication into the computer  21 . 
     (Step S 22  YES→S 23 ) 
     When the testing button B 2  is selected, the computer  21  transmits information on the scanning line L 1  and that on the scanning speed B 1  which were displayed at that moment to the controller  20  respectively as information on the set line and that on the set speed set by the user, together with the testing indication. 
     (Step S 11  Yes→S 12 ) 
     Upon receiving the information on the set line and the set speed and the testing indication, the CPU  201  of the controller  20  performs a setting of the galvanometer scanner  11  in accordance with the information. 
     Concretely, the CPU  201  resolves the set line into a plurality of unit vectors as shown in  FIG. 5(   a ). A size of the unit vector corresponds to an increasing function of the set speed. The CPU  201  generates the X-driving waveform ( FIG. 5(   b )) by converting X-components of the resolved set line into voltage values with a predetermined transfer characteristic, and generates the Y-driving waveform ( FIG. 5(   c )) by converting Y-components of the resolved set line into voltage values with a predetermined transfer characteristic. The predetermined transfer characteristics are characteristics previously determined by taking response characteristics of the galvanometer mirrors  111 X and  111 Y into consideration. Further, the CPU  201  stores the generated information on the X-driving waveforms and the Y-driving waveforms in the memory  202 A of the scanner controlling part  202 . 
     (Step S 13 ) 
     The CPU  201  of the controller  20  gives indications to the scanner controlling part  202  under the aforementioned setting conditions to thereby drive the galvanometer scanner  11 . However, since the CPU  201  does not drive any of the laser unit  1  and the light detector  19  at this time, there is no chance for the laser light to be incident on the sample  16 . 
     Further, during the driving of the galvanometer scanner  11 , the X-position signals and the Y-position signals output from the galvanometer scanner  11  are sampled by the scanner controlling part  202  and sequentially stored in the memory  202 A. The stored signals indicate the actual scanning line formed by the galvanometer scanner  11  (a scanning line of laser light when the galvanometer scanner  11  is driven while irradiating the laser light under the same setting condition). Hereinafter, the actual scanning line measured as above is referred to as “measured line” to distinguish it from the set line. 
     Steps S 12  and  13  described above correspond to the test. 
     (Step S 14 ) 
     The CPU  201  of the controller  20  calculates the measured line by reading the X-position signals and the Y-position signals stored in the memory  202 A and converting them into coordinates on the image I, and transmits information on the measured line to the computer  21 . 
     (Step S 24  Yes→S 25 ) 
     Upon receiving the information on the measured line, the computer  21  displays a measured line L 2  together with the scanning line L 1  on the setting screen as shown in  FIG. 6 . Through the display, the user can intuitively recognize a deviation between the scanning line L 1  input by himself/herself and the measured line L 2 . 
     For instance, when the scanning line L 1  is relatively steep and the scanning speed B 1  is relatively fast, the measured line L 2  is curved more gently than the scanning line L 1 . 
     (Step S 26  NO→S 22 ) 
     When the user is not satisfied with the measured line L 2 , the user is allowed to redraw the scanning line L 1  to have a gentle curve or change the scanning speed B 1  to a low speed side, and then, select the test button B 2  again. When the test button B 2  is selected, the aforementioned test is repeated. 
     (Step S 26  Yes→S 27 ) 
     When the user is satisfied with the measured line L 2 , the user just had to select the real scanning button B 3 . When the real scanning button B 3  is selected, the computer  21  transmits the information on the scanning line L 1 , the scanning speed B 1  and the laser intensity BO which were displayed at that moment to the controller  20  as the information on the set line, the set speed and set intensity set by the user, together with the real scanning indication. 
     (Step S 115  Yes, S 16 ) 
     Upon receiving the information on the set line, the set speed and the set intensity and the real scanning indication, the CPU  201  of the controller  20  performs settings of the laser unit  1 , the galvanometer scanner  11  and the light detector  19  in accordance with these pieces of information. Incidentally, if values of the set line and the set speed are the same as those of the last time, the setting of the galvanometer scanner  11  is omitted. 
     (Step S 17 ) 
     Under the aforementioned setting conditions, the CPU  201  of the controller  20  gives indications to the laser controlling part  207 , the detector controlling part  208  and the scanner controlling part  202 , to thereby synchronously drive the laser unit  1 , the light detector  19  and the galvanometer scanner  11  to obtain observation information. The obtainment of the observation information is continuously and repeatedly performed at a plurality of times, for instance. The above-described steps S 16  and  17  correspond to the real scanning. 
     (Step S 18 ) 
     The CPU  201  of the controller  20  transmits the observation information obtained in the real scanning to the computer  21  together with information on scanning conditions at the time of real scanning and the like. 
     (Step S 28  Yes→S 29 ) 
     Upon receiving the observation information, the computer  21  displays the observation information on the monitor  22  as shown in  FIG. 7 , for example.  FIG. 7  is a view in which respective pieces of scanning line information obtained by performing a laser scanning from a start point P 1  to an end point P 2  of the measured line L 2  from time t 0  to tn at a plurality of times are arranged lengthwise in time series. According to such a display, it is apparent that a part in which a reaction is detected (black-out part) gradually shifts from P 1  to P 2  (the above description corresponds to step S 29 ). 
     As described above, in the test (steps S 12  and  13 ) of the present system, only the galvanometer scanner  11  is driven under the scanning conditions designated by the user without irradiating laser light and the actual scanning line (measured line) at that time is measured. Accordingly, in this test, it is possible to obtain information on the measured line while preventing color fading and damage of the sample  16 . 
     Subsequently, the measured line L 2  is displayed on the monitor  22  after the test (refer to  FIG. 6 ), so that the user can determine whether the scanning conditions set by himself/herself are good or bad, and can give desired indications such as a change in the scanning conditions and executions of a real scanning and a retest, to the present system. 
     Further, in the present system, the scanning line L 1  set by the user is displayed together with the measured line L 2  (refer to  FIG. 6 ), so that the user can intuitively recognize a deviation between the both lines. 
     [Second Embodiment] 
     Hereinafter, a second embodiment will be described. The present embodiment is an embodiment of a confocal fluorescence laser scanning microscope system. Here, only a point of difference between this embodiment and the first embodiment will be described. The point of difference is that an optimizing function which automatically corrects details of the scanning conditions is mounted. 
     For this reason, operations shown in  FIG. 8  and  FIG. 9  are added to the operations of the controller  20  and the computer  21  of the present system, and an optimizing button B 4  is arranged on the setting screen as shown in  FIG. 10 . Further, on the setting screen, a region into which the user inputs a desired optimization margin B 5  is also provided. The optimization margin refers to a tolerance of deviation between the measured line after the optimization and the set line, and is expressed by, for instance, the number of pixels on the image I, or the like. 
     Hereinafter, the operations shown in  FIG. 8  and  FIG. 9  are specifically described. 
     (Step S 41  YES→S 42 ) 
     When the optimizing button B 4  is selected, the computer  21  transmits information on the scanning line L 1 , the scanning speed B 1  and the optimization margin B 5  which were displayed at that moment to the controller  20  as the information on the set line, the set speed and set margin set by the user, together with an optimizing indication. 
     (Step S 31 ) 
     Upon receiving the information on the set line, the set speed and the set margin and the optimizing indication, the CPU  201  of the controller  20  determines whether the test (steps S 12  and S 13  in  FIG. 3 ) with the set line and the set speed is already executed or not. 
     (Step S 32  NO→S 33 ) 
     If the test is not yet executed, the CPU  201  of the controller  20  executes a test with the set line and the set speed. This test is conducted in the same manner as in steps S 12  and S 13 . 
     (Step S 32  Yes) 
     If the test is already executed, the CPU  201  of the controller  20  skips step S 33  and proceeds to step S 34 . 
     (Step S 34 ) 
     The CPU  201  of the controller  20  calculates the measured line by reading the X-position signals and the Y-position signals stored in the memory  202 A at that moment and converting them into coordinates on the image I. 
     Further, the CPU  201  of the controller  20  subtracts the measured line from the set line set by the user, thereby calculating a difference between the both lines. At this time, the CPU  201  resolves each of the measured line and the set line into a plurality of unit vectors and then calculates a difference in X-direction and a difference in Y-direction, respectively, by each unit vector. A size of the unit vector corresponds to an increasing function of the set speed, and is the same as the one used when generating the driving waveform of the galvanometer scanner  11 . 
     (Step S 35 ) 
     The CPU  201  of the controller  20  determines whether or not a magnitude of the calculated difference falls within the set margin set by the user. 
     (Step S 35  NO→S 36 ) 
     If the magnitude of difference does not fall within the set margin, the CPU  201  of the controller  20  obtains a correction amount ΔV X (t) of the X-driving waveform by converting the difference in X-direction into a voltage value using a predetermined transfer characteristic. Further, the CPU  201  obtains a correction amount ΔV Y (t) of the Y-driving waveform by converting the difference in Y-direction into a voltage value using a predetermined transfer characteristic. Note that the predetermined transfer characteristics are the same as those used when generating the driving waveforms of the galvanometer scanner  11 . 
     Further, the CPU  201  adds the correction amount ΔV X (t) to the X-driving waveform stored in the memory  202 A at that moment, thereby correcting the X-driving waveform. Further, the CPU  201  adds the correction amount ΔV Y (t) to the Y-driving waveform stored in the memory  202 A at that moment, thereby correcting the Y-driving waveform. 
     (Step S 37 ) 
     The CPU  201  of the controller  20  determines whether or not changed frequencies of the X-driving waveform and the Y-driving waveform after the correction fall within a limiting frequency of the galvanometer scanner  11 . Here, the limiting frequency is determined based on the set speed set by the user and response characteristics of the galvanometer mirrors  111 X and  111 Y, and it becomes small as the set speed becomes faster. 
     (Step S 37  NO→S 38 ) 
     If the changed frequencies of the X-driving waveform and the Y-driving waveform after the correction do not fall within the limiting frequency, the set speed is lowered by one stage, and thereafter, the procedure goes back to step S 34  and a calculation of difference is performed again. 
     (Step S 37  Yes→S 33 ) 
     If the changed frequencies of the X-driving waveform and the Y-driving waveform after the correction fall within the limiting frequency, the procedure goes back to step S 33 , and a retest is conducted using the corrected X-driving waveform and Y-driving waveform. The above-described steps S 33  through S 38  correspond to the optimization. 
     (Step S 35  YES→S 39 ) 
     Thereafter, if the magnitude of difference calculated in step S 34  falls within the set margin, the CPU  201  of the controller  20  terminates the optimization, and transmits information on the set speed and the measured line after optimization to the computer  21 . Note that the calculation method of the measured line is the same as described above. 
     (Step S 43  YES→S 44 ) 
     Upon receiving the information on the set speed and the measured line after optimization, the computer  21  reflects these pieces of information upon the setting screen as shown in  FIG. 11 , for instance. In  FIG. 11 , what is indicated by the reference numeral B 1 ′ is the set speed after optimization and what is indicated by the reference numeral L 2 ′ is the measured line after optimization. Accordingly, the user can recognize a result of the optimization. 
     (Step S 45  Yes→S 48 ) 
     After that, when the real scanning button B 3  is selected, the computer  21  transmits information on the scanning line L 1 , the scanning speed B 1 ′, the laser intensity B 0  and the optimization margin B 5  which were displayed at that moment to the controller  20  as the information on the set line, the set speed, the set intensity and the set margin set by the user, together with the real scanning indication. 
     (Step S 301  YES→S 302 ) 
     Upon receiving the information on the set line, the set speed, the set intensity and the set margin and the real scanning indication, the CPU  201  of the controller  20  determines whether the optimization with the set line, the set speed and the set margin is already executed or not. 
     (Step S 302  No→S 303 ) 
     If the optimization is not yet executed, the CPU  201  of the controller  20  performs settings of the laser unit  1 , the light detector  19  and the galvanometer scanner  11  in accordance with the set line, the set speed and the set intensity. 
     (Step S 302  Yes) 
     Meanwhile, if the optimization is already executed, the CPU  201  of the controller  20  performs settings of only the laser unit  1  and the light detector  19  in accordance with the set line, the set speed and the set intensity, and stores the set contents (driving waveforms) of the galvanometer scanner  11  while keeping them as they were optimized. 
     (Step S 305 ) 
     Under the aforementioned setting conditions, the CPU  201  of the controller  20  gives indications to the laser controlling part  207 , the detector controlling part  208  and the scanner controlling part  202 , to thereby synchronously drive the laser unit  1 , the light detector  19  and the galvanometer scanner  11  to obtain observation information. The obtainment of the observation information is continuously and repeatedly performed at a plurality of times, for instance. The above-described steps S 303 ,  304  and  305  correspond to the real scanning. 
     (Step S 306 ) 
     The CPU  201  of the controller  20  transmits the observation information obtained in the real scanning to the computer  21  together with the scanning conditions at the time of real scanning and the like. 
     (Step S 49  Yes→S 50 ) 
     Upon receiving the observation information, the computer  21  displays the observation information on the monitor  22 . A display method at this time is the same as that shown in  FIG. 7 , for instance (the above description corresponds to step S 50 ). 
     As described above, in the optimization of the present system (steps S 33  through S 38 ), the test and the correction of the driving waveforms are repeatedly conducted until the difference between the measured line and the set line falls within the set margin (until the determination of step S 35  becomes YES) (step S 36 ). By the repetition, values of the driving waveforms gradually become close to optimal values. Further, in this optimization, the set speed is changed to the low-speed side according to demand, to thereby obtain the optimal values. Therefore, according to the optimization of the present system, details of the scanning conditions are automatically optimized. 
     Note that in step S 34  of the present system, the difference between the measured line and the set line is calculated and then the difference is converted into the voltage value, but, it is possible that the measured line and the set line are converted into the voltage values and then the difference between the both is calculated. However, in such a case, it becomes necessary to convert a judgment standard (set margin) in step S 35  into the voltage value, and the conversion in step S 36  becomes unnecessary. 
     Further, in the optimization of the present system (steps S 33  through S 38 ), the test and the correction of the driving waveforms are repeatedly conducted until the difference between the measured line and the set line falls within the set margin (until the determination of step S 35  becomes YES), but, they may be repeatedly conducted at previously determined number of times. Further, the number of repetition may be designated by the user. 
     Further, in the real scanning of the present system (steps S 303  through S 305 ), the driving waveforms after optimization are automatically adopted when the optimization is already executed, but, it is also possible to get the user to select whether the driving waveforms after optimization are adopted or driving waveforms which are regenerated are adopted, and then to follow a result of the selection. 
     Further, the controller  20  and the computer  21  of the present system may be operated as follows after the optimization. 
     The CPU  201  of the controller  20  transmits information on the driving waveforms after optimization to the computer  21 . The computer  21  stores the received information on the driving waveforms in accordance with an indication from the user. At this time, the information on the driving waveforms is corresponded to information on the image I of the sample  16 . Thereafter, when an indication to recall the driving waveforms is made from the user, the stored information on the driving waveforms is read and transmitted to the controller  20 . In such a case, the controller  20  writes the received driving waveforms into the memory  202 A to thereby perform a setting of the galvanometer scanner  11 . According to such operations, the number of executions of the processing regarding the optimization can be kept to the minimum. Further, the user can use the driving waveforms after optimization by recalling them at a desired timing. 
     Further, in the first and second embodiments, examples in which only the galvanometer scanner  11  is driven under the state where no laser is irradiated from the laser unit  1  when calculating the measured line are shown, but, the present invention is not limited to this and it is possible that the galvanometer scanner  11  is driven under the state where laser having an intensity lower than that of laser light irradiated in the real scanning is irradiated from the laser unit  1 . It is possible to suppress the color fading and damage of the sample  16  only by setting an intensity of laser light to one being lower than that of the laser light used when performing a real scanning as described above. 
     [Other Features] 
     Note that the aforementioned microscope body  100  is a laser scanning microscope having both the function of fluorescence detection and the function of confocal detection, but, the present invention is also applicable to a laser scanning microscope which does not have either or both of the function of fluorescence detection and the function of confocal detection. Further, the present invention can be also applied to a laser scanning apparatus which does not have the detection function. 
     The many features and advantages of the embodiments are apparent from the detailed specification and, thus, it is intended by the appended claims to cover all such features and advantages of the embodiments that fall within the true spirit and scope thereof. Further, since numerous modifications and changes will readily occur to those skilled in the art, it is not desired to limit the inventive embodiments to the exact construction and operation illustrated and described, and accordingly all suitable modifications and equivalents may be resorted to, falling within the scope thereof.