Microscope and method for controlling the microscope

Abstract

The microscope comprises a light source that can directly control an ON state in which coherent light is emitted and an OFF state in which no light is emitted; an illumination optical system that irradiates light to form an illumination area on a sample; an optical path changing member that changes the optical path of the light; and a control unit that controls the light source and the optical path changing member. The control unit controls the optical path changing member to switch between a first state in which, when the light source is in the ON state, light forms the illumination area on the sample, and when it is in the OFF state, no illumination area is formed, and a second state in which, regardless of whether the light source is in the ON or OFF state, light does not form an illumination area on the sample. In the second state, the control unit controls the light source to be in the ON state.

Description

Technical Field

[0001] The present invention relates to a microscope and a method for controlling the microscope.

Background Art

[0002] Patent Document 1 describes a confocal scanning microscope that scans a sample with irradiation laser light. [Prior Art Document] [Patent Document] [Patent Document 1] International Publication No. WO2009 / 011441 [General Disclosure]

[0003] In a first aspect of the present invention, a microscope is provided. The microscope includes a light source that can be directly controlled between an ON state in which coherent light is emitted and an OFF state in which the light is not emitted, an illumination optical system that irradiates the light to form an illumination region on a sample, an optical path changing member that changes the optical path of the light, and a control unit that controls the light source and the optical path changing member. The control unit controls the optical path changing member such that when the light source is in the ON state, the light forms the illumination region on the sample, and when in the OFF state, the illumination region is not formed (a first state), and switches between a second state in which the light does not form the illumination region on the sample regardless of whether the light source is in the ON state or the OFF state, and in the second state, controls the light source to be in the ON state.

[0004] In the second state, the control unit may control the light source so that the ON state and the OFF state are repeated at a predetermined timing.

[0005] During at least one of the switching from the first state to the second state and the switching from the second state to the first state, the control unit may control the light source to be in the OFF state.

[0006] The illumination optical system further includes a focusing lens, wherein in the first state, the light passes through the focusing lens, and in the second state, the light does not pass through the focusing lens and may be incident at a predetermined standby position.

[0007] The system further includes a holding member for holding the light-gathering lens, and the standby position may be a part of the holding member.

[0008] A light-shielding member may be provided in part of the above.

[0009] The present invention may further include a reflective member provided at the standby position that reflects the light, and an absorbing member that absorbs at least a portion of the light reflected from the reflective member.

[0010] The optical path changing member may be a galvanometer mirror or a spatial light modulator.

[0011] The control unit may, in the second state, turn the light source to the ON state, wait for a predetermined time, and then switch the light back to the first state.

[0012] The predetermined time may be the time during which the output value from the light source stabilizes.

[0013] A second embodiment of the present invention provides a method for controlling a microscope. The method for controlling a microscope includes a light source that can directly control an ON state in which coherent light is emitted and an OFF state in which the light is not emitted; an illumination optical system that irradiates the sample with the light to form an illumination region; an optical path changing member that changes the optical path of the light; and a control unit that controls the light source and the optical path changing member, wherein the method for controlling a microscope includes a control step of controlling the optical path changing member to switch between a first state in which, when the light source is in the ON state, the light forms the illumination region on the sample and when it is in the OFF state, the light does not form the illumination region on the sample, and a second state in which, regardless of whether the light source is in the ON state or the OFF state, the light does not form the illumination region on the sample, and in the second state, the control step of setting the light source to the ON state.

[0014] It should be noted that the above summary of the invention does not list all the necessary features of the present invention. Furthermore, subcombinations of these features may also constitute an invention. [Brief explanation of the drawing]

[0015] [Figure 1] The schematic configuration of the microscope 200 in the first embodiment is shown. [Figure 2] The schematic configuration of the scan head 100 in the first embodiment is shown. [Figure 3] An example of scanning by the scan head 100 in the first embodiment is shown. [Figure 4] This shows the ON / OFF timing chart of the laser light source 101 in the first embodiment. [Figure 5] This is a flowchart showing the operation of the scan head 100 in the first embodiment. [Figure 6] The schematic configuration of the scan head 110 in the second embodiment is shown. [Figure 7] The schematic configuration of the scan head 120 in the third embodiment is shown. [Figure 8] (a) and (b) show other control examples of the ON / OFF state of the laser light source 101. [Figure 9] An example of computer 2200 is shown. [Modes for carrying out the invention]

[0016] The present invention will be described below through embodiments of the invention. The following embodiments are not intended to limit the invention as claimed. Not all combinations of features described in the embodiments are necessarily essential to the solution of the invention.

[0017] Figure 1 shows a schematic configuration of the microscope 200 in the first embodiment. As shown in Figure 1, the microscope 200 in the first embodiment includes a scan head 100, a laser light source 101, a photodetector 102, a microscope body 103, and a control unit 104. The microscope 200 is a microscope that scans a laser, and may be, for example, a confocal microscope. The figure shows an xyz coordinate system.

[0018] The laser light source 101 emits laser light, which is excitation light, towards the sample 106. The laser light source 101 is configured to be switchable between an ON state (hereinafter simply referred to as ON) in which excitation light is emitted to the outside, and an OFF state (hereinafter simply referred to as OFF) in which excitation light is not emitted to the outside. Examples of the laser light source 101 include semiconductor lasers, gas lasers, and solid-state lasers. In other words, the control unit 104 directly controls the laser light source 101 itself to switch between ON and OFF. This is different from a laser light source unit including a laser light source and a mechanical shutter or AOTF, in which the laser light source continuously emits excitation light, and the mechanical shutter or AOTF switches between ON and OFF. The sample 106 to be observed is placed on the stage of the microscope body 103. The microscope body 103 has a first objective lens 105a and a second objective lens 105b (hereinafter collectively referred to as the objective optical system 105).

[0019] The photodetector 102 receives the fluorescence emitted from the sample 106 using a photomultiplier tube or similar device, converts it into an electrical signal, and outputs it to the control unit 104. The control unit 104 stores the input data in a computer, arranges it to form a single image, constructs the observed fluorescence image, and displays the image on a display. The control unit 104 is connected to each component of the microscope 200 and can control each component. For example, the control unit 104 can control the ON / OFF state of the laser light source 101, the operation of the scan head 100, or the stage of the microscope body 103.

[0020] Figure 2 shows the schematic configuration of the scan head 100 in the first embodiment. As shown in Figure 2, the scan head 100 in the first embodiment includes an input port 11, a collimating lens 12, a dichroic mirror 13, a galvanometer mirror 14, a scanning optical system 15, a lens barrel 16, a microscope port 17, a condenser lens 18, a pinhole 19, and an output port 20. Note that the scan head 100 also has other optical members such as a prism or a mirror that bend the optical path of the excitation light.

[0021] The input port 11 is a port for incidentally introducing the excitation light, which is the laser light from the laser light source 101, into the scan head 100. An optical fiber for the excitation light is connected to the input port 11. The collimating lens 12 converts the excitation light incident from the input port 11 into parallel light. The dichroic mirror 13 reflects the excitation light that has become parallel light by the collimating lens 12.

[0022] The galvanometer mirror 14 is an example of an optical path changing member that changes the optical path of the excitation light. The galvanometer mirror 14 is composed of a pair of mirrors, one mirror that rotates the excitation light around the x-axis and the other mirror that rotates the excitation light around the y-axis. The pair of galvanometer mirrors 14 reflects the angle of the excitation light reflected by the dichroic mirror 13 in two arbitrarily orthogonal biaxial directions. Therefore, the galvanometer mirror 14 can scan the excitation light on the xy plane.

[0023] The scanning optical system 15 is a focusing lens that focuses the excitation light reflected by the galvanometer mirror 14. The scanning optical system 15 has an incident surface 15a into which the excitation light is incident. The lens barrel 16 is a holding member that holds the scanning optical system 15, which is a focusing lens. The lens barrel 16 has a side surface 16a parallel to the xy plane. The side surface 16a is located outside the incident surface 15a. A standby position 107 is provided on the side surface 16a of the lens barrel 16 for waiting for excitation light from the laser light source 101. In this embodiment, the excitation light is input to the objective optical system 105 located downstream of the optical path via the microscope port 17. The scanning optical system 15 and the objective optical system 105 function as an illumination optical system that focuses the laser light to form an illumination area on the sample 106.

[0024] Excitation light incident on the scanning optical system 15 from the incident surface 15a is focused by the scanning optical system 15 and output from the microscope port 17, irradiating the sample 106 via the objective optical system 105, etc. On the other hand, excitation light incident on a location other than the incident surface 15a of the scanning optical system 15, for example, on the side surface 16a of the lens barrel 16, is not output to the microscope port 17. Therefore, excitation light incident on the standby position 107 provided on the side surface 16a of the lens barrel 16 is not output from the microscope port 17 and is not irradiated to the sample 106.

[0025] As described above, the excitation light emitted from the laser light source 101 passes through the input port 11, collimating lens 12, dichroic mirror 13, galvanometer mirror 14, incident surface 15a of the scanning optical system 15, microscope port 17, and objective optical system 105 in that order, and is irradiated onto the sample 106. Upon receiving the irradiated excitation light, the sample 106 generates fluorescence at a specific wavelength from the excited fluorescent dye. The fluorescence, which is the observation light, passes through the objective optical system 105 of the microscope body 103 and is incident on the scan head 100 from the microscope port 17.

[0026] The incident fluorescence passes through the microscope port 17, scanning optical system 15, galvanometer mirror 14, and dichroic mirror 13, and is then converted into focused light by the focusing lens 18. The pinhole 19 is positioned conjugate to the focal position of the objective lens 105a, shielding the area outside the conjugate position. The output port 20 is a port for outputting the fluorescence that has passed through the pinhole 19 to the photodetector 102. The fluorescence generated in the sample 106 is output from the output port 20 and input to the photodetector 102.

[0027] As described above, the fluorescence emitted from the sample 106 passes through the objective optical system 105, microscope port 17, scanning optical system 15, galvanometer mirror 14, dichroic mirror 13, focusing lens 18, pinhole 19, and output port 20 in that order, and is finally input to the photodetector 102. In the photodetector 102, the fluorescence emitted from the sample 106 is received by a photomultiplier tube or the like, converted into an electrical signal, and output to the control unit 104.

[0028] The control unit 104 controls the galvanometer mirror 14, which is an optical path changing member, to switch between a first state in which the laser light source 101 forms an illumination area on the sample 106 when it is ON, and does not form an illumination area when it is OFF, and a second state in which the laser light does not form an illumination area on the sample 106 regardless of whether the laser light source 101 is ON or OFF, and in the second state, it controls the laser light source 101 to be ON. Specifically, the control unit 104 controls the galvanometer mirror 14, which is an optical path changing member, to switch between a first optical path (optical path in the first state) in which the excitation light is output from the microscope port 17, and a second optical path (optical path in the second state) in which the excitation light is not output from the microscope port 17. In Figure 2, the control unit 104 controls the excitation light to be incident on the incident surface 15a of the scanning optical system 15 as the first optical path. The control unit 104 controls the excitation light to enter the standby position 107 on the side surface 16a of the lens barrel 16 as the second optical path. The first optical path can be described not as a single optical path, but rather as a collection of multiple optical paths when scanning over the sample 106 in the XY direction.

[0029] Figure 3 shows an example of excitation light scanning by the scan head 100 in the first embodiment. Figure 3 shows the observable range (maximum observable range) 108 of the objective optical system 105 of the microscope 200. The observable range 108 in Figure 3 is the range on the sample 106. The observable range 108 on the sample 106 changes depending on the magnification of the objective lens 105a. In this embodiment, excitation light scanning is a scan along multiple lines within the observable range 108 of the objective optical system 105. In Figure 3, multiple lines, labeled as the 1st line, 2nd line, ... nth line, are shown as thick solid lines. Scanning is performed sequentially from left to right along the thick solid lines showing the multiple lines in Figure 3. The leftmost point of each of the multiple lines is also called the scanning start position. Figure 3 also shows a standby position 107. The standby position 107 in Figure 3 schematically shows the standby position 107 provided on the side surface 16a of the lens barrel 16 in Figure 2. Furthermore, optically, the side surface 16a of the lens barrel 16 corresponds to the outside of the observable range 108 of the objective optical system 105.

[0030] In the first embodiment, when the microscope 200 system is started, the galvanometer mirror first moves (rotates by a predetermined angle) so that the excitation light emitted from the laser light source 101 is directed towards the standby position 107 in Figure 3. After the galvanometer mirror has moved, the laser light source 101 is turned ON and the excitation light is irradiated onto the standby position 107, and the system waits until the output value of the laser light source 101 stabilizes. The excitation light irradiated onto this standby position 107 does not irradiate the sample 106.

[0031] Depending on the type of laser light source 101, after turning on the laser light source 101 and the output value (intensity of excitation light) reaches a preset value, it may take several milliseconds to several seconds for that output value to actually stabilize. If the laser light source 101 takes several milliseconds to stabilize, there is no problem in waiting for those few milliseconds until the output value stabilizes. On the other hand, if a laser light source 101 takes several seconds to stabilize, it becomes problematic if the laser light source 101 is completely turned off and then turned on again, and then a few seconds are waited for the output value to stabilize each time the observation position of the sample 106 is changed or the sample 106 is replaced, because the sequentiality of the observation is lost. Furthermore, if the waiting is not performed, the brightness of the observed image may change unexpectedly, making quantitative analysis impossible.

[0032] In contrast, in this embodiment, before irradiating the sample 106 with excitation light, the excitation light is set to irradiate the standby position 107, then the laser light source 101 is turned ON and the system waits until the output value from the laser light source 101 stabilizes. Therefore, the irradiation of the sample 106 with excitation light can be started when the output value of the laser light source 101 is stable, solving the above problem, and the brightness of the images obtained from each capture can be acquired, displayed, and saved in a stable state. Note that immediately after system startup, after setting the excitation light to irradiate the standby position 107, the laser light source 101 is turned ON and the system waits for several seconds to allow the output value of the laser light source 101 to stabilize. The need to wait for several seconds is only required once immediately after starting the microscope 200, and thereafter, unless the laser light source 101 is turned OFF, a waiting time of several seconds is not required.

[0033] At the timing to start scanning the excitation light, the excitation light is scanned from the standby position 107 to the scanning start position, thereby moving the irradiation position of the excitation light. Note that the upper left point 301 of the first line in Figure 3 is the scanning start position, and the coordinates of point 301 are (Sx, Sy). The lower right point 306 is the scanning end position, and the coordinates of point 306 are (Sx+m-1, Sy+n-1). m is the amount of movement in the X direction (m is an integer of 1 or more), and n is the amount of movement in the Y direction (number of lines) (n is an integer of 1 or more). The coordinates of the standby position 107 are (a, b). These coordinates of the standby position 107 are the position when projected directly onto the sample 106 without passing through the objective optical system 105. Here, a and b are values ​​that are outside the observable range 108 of the objective optical system 105. Here, the laser light source 101 is turned OFF during the short time it takes to scan the excitation light from the standby position 107 to the scanning start position 301.

[0034] When the excitation light has finished moving to the scanning start position 301, the laser light source 101 is turned ON again, and the excitation light is scanned for one line in the +x direction to move the irradiation position of the excitation light to point 302 (Sx+m-1, Sy) on the right side of the first line, and observation data of the sample 106 is acquired. This observation data is stored in the memory of the control unit 104.

[0035] When the excitation light irradiation position reaches point 302 (Sx+m-1, Sy) on the right side of the first line, the laser light source 101 is turned OFF, and the excitation light irradiation position is reversed to move to point 303 (Sx, Sy+1) on the left side of the second line. The laser light source 101 is kept OFF until it reaches point 303 (Sx, Sy+1). In this way, as described above, extra light is not irradiated onto the sample 106 during the reversal operation where data is not accumulated, thereby reducing phototoxicity and damage to the sample 106. Thereafter, scanning is performed similarly up to the 3rd line, 4th line, ... nth line.

[0036] When the final line, the nth line, is reached, at scanning end position 306 (Sx+m-1, Sy+n-1), the excitation light is moved from scanning end position 306 to standby position 107. During the short time it takes for the excitation light to move from scanning end position 306 to standby position 107, the laser light source 101 is turned OFF. After the excitation light has finished moving to standby position 107, the laser light source 101 is turned ON again and left to wait for a while to stabilize its output value.

[0037] As described above, in order to suppress phototoxicity and damage to sample 106, the laser light source 101 is briefly turned OFF while the excitation light moves from the standby position 107 to the scanning start position 301, during the reversal between points such as point 302 to point 303, and while moving from the scanning end position 306 to the standby position 107. Although the output value of the laser light source 101 may fluctuate slightly due to the brief OFF period, the effect on the output value of the laser light source 101 is limited because the OFF period is short. Furthermore, if a sample 106 that is less susceptible to phototoxicity or damage, or a sample 106 that is not susceptible to phototoxicity or damage, is used, it is not necessary to turn OFF the laser light source 101 during the above periods. Additionally, the laser light source 101 may be turned OFF for only one or two of the above three movement periods.

[0038] Figure 4 shows a timing chart for the ON / OFF control of the laser light source 101 in the first embodiment. The three solid lines in Figure 4, from top to bottom, represent the x-coordinate and y-coordinate of the excitation light irradiation position, and the ON / OFF control of the laser light source 101. The x and y coordinates in Figure 4 are the same as those in Figure 3. At time t0 when the system is started, the excitation light irradiation position is at the initial position (location is undefined). Then, at time t1, the irradiation position moves to the standby position 107 (a, b). This is outside the observable range 108 of the objective optical system 105, and even if the laser light source 101 is output at this position, it will not irradiate the sample 106. For a predetermined time from time t1 to time t2, the laser light source 101 is turned ON and waits at the standby position 107 until the output value of the laser light source 101 stabilizes.

[0039] At time t2, the laser light source 101 is turned OFF, and the excitation light irradiation position is moved to point 301 (Sx, Sy), which is the scanning start position. Then, at time t3, the laser light source 101 is turned ON and scanning begins. After turning ON the laser light source 101, the excitation light irradiation position is scanned to point 302 (Sx+m-1, Sy) on the right side of the first line. During this time, the y-coordinate (Sy) of the excitation light irradiation position is maintained. Then, at time t4, the laser light source 101 is turned OFF, and the excitation light irradiation position is reversed to move to the starting point 303 of the second line, and the laser light source 101 is turned ON again to perform scanning of the second line. Scanning continues thereafter until the nth line is completed.

[0040] Subsequently, at time t6, when the excitation light irradiation position reaches point 306 (Sx+m-1, Sy+n-1), which is the scanning end position, the laser light source 101 is turned OFF and moved to standby position 107 (a, b). After the move is complete, at time t7, the laser is turned ON and put into standby mode, waiting for the command for the next scan.

[0041] Figure 5 is a flowchart showing the operation of the scan head 100 in the first embodiment. In step S01, after the system is started, the control unit scans the galvanometer mirror 14 to move the excitation light irradiation position to the standby position 107. After moving to the standby position 107, in the next step S02, the laser light source 101 is turned ON, and in the next step S03, the microscope 200 waits for a few seconds only once immediately after startup until the output value of the laser light source 101 stabilizes.

[0042] In the next step S04, when the user instructs to start scanning, in the next step S05, the laser light source 101 is turned OFF, and in the next step S06, the irradiation position of the excitation light is moved to the scanning start position (first, the scanning start position 301 (Sx, Sy) of the first line). In the next step S07 after the move, the laser light source 101 is turned ON, and in the next step S08, one line is scanned. After that, in the next step S09, the laser light source 101 is turned OFF, the irradiation position of the excitation light is reversed, and the scanning continues sequentially to the next line, up to the nth line which is the final line.

[0043] In the next step S10, if the excitation light irradiation position reaches point 306 (Sx+m-1, Sy+n-1), which is the scanning end position (YES in step S10), in the next step S11, the laser light source 101 is turned OFF and the excitation light irradiation position is moved to the standby position 107. In the next step S12 after the move, the laser light source 101 is turned ON and left to wait until the output value stabilizes. In the next step S13, if there is no longer a scanning range remaining and the scanning is complete, the experiment is terminated (YES in step S13). If there is still a scanning range remaining (NO in step S13), the process returns to step S04 and the above process is repeated.

[0044] In the microscope 200 of the first embodiment, a standby position 107 is provided where the excitation light is waiting. The laser light source 101 is turned ON at the standby position 107 and left waiting until the output value stabilizes. Once the output value is stable, irradiation of the sample 106 is started. This eliminates the problem of the acquired image brightness undergoing unexpected changes, making quantitative analysis impossible, and stabilizes the brightness of the acquired image.

[0045] In the first embodiment of the microscope 200, the laser light source 101 is turned OFF while moving from the standby position 107 to the scanning start position 301, during the reversal from point 302 to point 303, and while moving from the scanning end position 306 to the standby position 107. This reduces phototoxicity and damage to the sample 106 caused by irradiation with excitation light.

[0046] Figure 6 shows a schematic configuration of the scan head 110 in the second embodiment. In Figure 6, components identical to those in the scan head 100 in the first embodiment shown in Figure 2 are denoted by the same reference numerals and their descriptions are omitted. As shown in Figure 6, the scan head 110 in the second embodiment is provided with a light-shielding means 21 at the standby position 107 where the excitation light is kept on standby. By providing the light-shielding means 21 at the standby position 107, the generation of secondary light due to the reflection of excitation light at the standby position, which may occur if the light-shielding means 21 is not provided, is suppressed. This prevents stray light based on such secondary light from irradiating the sample 106 through the microscope port 17.

[0047] Figure 7 shows a schematic configuration of the scan head 120 in the third embodiment. In Figure 7, components identical to those in the scan head 100 in the first embodiment shown in Figure 2 are denoted by the same reference numerals and their descriptions are omitted. As shown in Figure 7, in order to prevent stray light, the scan head 120 in the third embodiment has a reflective member 22 in the middle of the path of the excitation light toward the standby position 107, instead of a standby position 107 for waiting for the excitation light to wait, and a beam dump 23 that absorbs the excitation light is provided at the destination of the excitation light reflected by the reflective member 22.

[0048] The reflective member 22 has the function of separating the excitation light directed toward the standby position 107 from the excitation light directed toward the incident surface 15a, which is the principal beam. The beam dump 23 has the function of further absorbing at least a portion of the excitation light separated from the principal beam by the reflective member 22 by mechanical or optical means. By installing the reflective member 22 in the path to the standby position 107 and combining it with the beam dump 23, the generation of secondary light can be suppressed, and stray light can be prevented from irradiating the sample 106 through the microscope port 17.

[0049] Figure 8 shows another example of ON / OFF control of the laser light source 101. Figure 8(a) shows a portion of the timing chart shown in Figure 4. In Figure 4, while the excitation light is waiting in standby position 107, the laser light source 101 is controlled to be OFF at time t0, ON from time t1 to time t2, OFF at time t2, and then ON again at time t3. As shown in Figure 8(a), overall, the laser light source 101 is controlled to be ON and OFF at a time ratio of approximately 3:1 to 4:1.

[0050] Furthermore, in the embodiment shown in Figure 4, the laser light source 101 was controlled to be constantly ON while the excitation light was waiting at the standby position 107. However, while the excitation light was waiting at the standby position 107, the ON / OFF state of the laser light source 101 may be controlled as shown in Figure 8(b). Figure 8(b) shows another control example, illustrating in detail the time t1 to time t2 while the excitation light was waiting at the standby position 107.

[0051] As shown in Figure 8(b), in other control examples, instead of keeping the laser light source 101 constantly ON during the second state in which the excitation light is waiting at the standby position 107, the laser light source 101 is alternately turned ON and OFF at the same timing as in Figure 8(a). As shown in Figure 8(b), the ON / OFF time ratio of the laser light source 101 is approximately 3:1 to 4:1, similar to Figure 8(a). In other words, the control in Figure 8(b) is similar to the control in Figure 8(a). By controlling the ON / OFF of the laser light source 101 as shown in Figure 8(b) while the laser light source 101 is waiting at the standby position 107, the output timing and output state can be made similar to the ON / OFF control during actual scanning, thereby further stabilizing the output value of the laser light source 101.

[0052] In the above embodiment, a galvanometer mirror 14 was used as the optical path changing member. However, instead of the galvanometer mirror 14, a DMD (Digital Mirror Device), a MEMS (Micro Electro Mechanical Systems) shutter, a spatial light modulator (SLM), etc., may be used as the optical path changing member.

[0053] In the above embodiment, the laser light source 101 was turned OFF to reduce phototoxicity and damage to the sample 106 while the excitation light was moving from the standby position 107 to the scanning start position 301, during the reversals from point 302 to point 303, and while moving from the scanning end position 306 to the standby position 107. However, instead, the laser light source 101 may be temporarily shut off using a mechanical shutter with a motor or the like. In this case, scanning may be performed taking into account the opening and closing speed of the mechanical shutter.

[0054] In the above embodiment, the excitation light switches between a first optical path that passes through the scanning optical system 15 located downstream of the galvanometer mirror 14 and a second optical path that does not pass through the scanning optical system 15, thereby switching between the first and second states. However, a mirror may be placed between the scanning optical system 15 and the objective optical system 105, and the mirror may be controlled to switch the excitation light between an optical path that passes through the objective optical system 105 and an optical path that does not pass through the objective optical system 105. Specifically, the standby position 107 may be placed outside the objective optical system 105 (similar to the side surface 16a of the lens barrel 16, on the side surface of the lens barrel of the second objective lens 105b), that is, in a position where the excitation light does not enter the objective optical system 105. Instead of the laser light source 101, a superluminescent diode (SLD) that can be directly controlled and switched ON or OFF by the control unit 104 may be used. In that case, a filter that allows only a predetermined wavelength of light emitted from the SLD to pass through may be placed to generate excitation light of a desired wavelength. Furthermore, the laser light source 101 and the SLD are light sources that emit coherent light.

[0055] Furthermore, various embodiments of the present invention may be described with reference to flowcharts and block diagrams, where a block may represent (1) a stage in a process in which an operation is performed or (2) a section of a device having the role of performing an operation. Specific stages and sections may be implemented by dedicated circuits, programmable circuits supplied with computer-readable instructions stored on a computer-readable medium, and / or processors supplied with computer-readable instructions stored on a computer-readable medium. Dedicated circuits may include digital and / or analog hardware circuits, and may include integrated circuits (ICs) and / or discrete circuits. Programmable circuits may include reconfigurable hardware circuits, including logical AND, logical OR, logical XOR, logical NAND, logical NOR, and other logic operations, flip-flops, registers, memory elements such as field-programmable gate arrays (FPGAs), programmable logic arrays (PLAs), etc.

[0056] Computer-readable media may include any tangible device capable of storing instructions to be executed by a suitable device, and as a result, computer-readable media having instructions stored therein will comprise a product containing instructions that can be executed to create means for performing operations specified in a flowchart or block diagram. Examples of computer-readable media may include electronic storage media, magnetic storage media, optical storage media, electromagnetic storage media, semiconductor storage media, etc. More specific examples of computer-readable media may include floppy disks, diskettes, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), electrically erasable programmable read-only memory (EEPROM), static random access memory (SRAM), compact disk read-only memory (CD-ROM), digital versatile disk (DVD), Blu-ray (RTM) disk, memory stick, integrated circuit card, etc.

[0057] Computer-readable instructions may include assembler instructions, instruction set architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state setting data, or source code or object code written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Smalltalk®, Java®, C++, and traditional procedural programming languages ​​such as the C programming language or similar programming languages.

[0058] Computer-readable instructions may be provided locally or via a wide area network (WAN), such as a local area network (LAN) or the internet, to a processor or programmable circuit of a general-purpose computer, a special-purpose computer, or other programmable data processing device, and these instructions may be executed to create means for performing operations specified in a flowchart or block diagram. Examples of processors include computer processors, processing units, microprocessors, digital signal processors, controllers, microcontrollers, and the like.

[0059] Figure 9 shows an example of a computer 2200 in which multiple aspects of the present invention may be embodied in whole or in part. A program installed on the computer 2200 can cause the computer 2200 to function as an operation or one or more sections of an apparatus according to an embodiment of the present invention, or to execute such operation or one or more sections, and / or to cause the computer 2200 to execute a process or a stage of such process according to an embodiment of the present invention. Such a program may be executed by the CPU 2212 to cause the computer 2200 to perform a particular operation associated with some or all of the blocks in the flowcharts and block diagrams described herein.

[0060] The computer 2200 according to this embodiment includes a CPU 2212, RAM 2214, a graphics controller 2216, and a display device 2218, which are interconnected by a host controller 2210. The computer 2200 also includes input / output units such as a communication interface 2222, a hard disk drive 2224, a DVD-ROM drive 2226, and an IC card drive, which are connected to the host controller 2210 via an input / output controller 2220. The computer also includes legacy input / output units such as a ROM 2230 and a keyboard 2242, which are connected to the input / output controller 2220 via an input / output chip 2240.

[0061] The CPU 2212 operates according to programs stored in the ROM 2230 and RAM 2214, thereby controlling each unit. The graphics controller 2216 retrieves image data generated by the CPU 2212 from a frame buffer provided in RAM 2214 or from itself, and displays the image data on the display device 2218.

[0062] The communication interface 2222 communicates with other electronic devices via a network. The hard disk drive 2224 stores programs and data used by the CPU 2212 in the computer 2200. The DVD-ROM drive 2226 reads programs or data from the DVD-ROM 2201 and provides them to the hard disk drive 2224 via the RAM 2214. The IC card drive reads programs and data from the IC card and / or writes programs and data to the IC card.

[0063] The ROM 2230 stores boot programs and / or programs that depend on the computer 2200's hardware, which are executed by the computer 2200 when activated. The input / output chip 2240 may also connect various input / output units to the input / output controller 2220 via parallel ports, serial ports, keyboard ports, mouse ports, etc.

[0064] The program is provided on a computer-readable medium such as a DVD-ROM 2201 or an IC card. The program is read from the computer-readable medium and installed on a hard disk drive 2224, RAM 2214, or ROM 2230, which are also examples of computer-readable medium, and executed by the CPU 2212. The information processing described within these programs is read by the computer 2200, resulting in coordination between the program and the various types of hardware resources described above. The apparatus or method may be configured to realize the manipulation or processing of information in accordance with the use of the computer 2200.

[0065] For example, when communication is performed between a computer 2200 and an external device, the CPU 2212 may execute a communication program loaded into RAM 2214 and, based on the processing described in the communication program, instruct the communication interface 2222 to perform communication processing. Under the control of the CPU 2212, the communication interface 2222 reads transmission data stored in a transmission buffer processing area provided in a recording medium such as RAM 2214, a hard disk drive 2224, a DVD-ROM 2201, or an IC card, transmits the read transmission data to the network, or writes received data received from the network to a reception buffer processing area provided on the recording medium.

[0066] Furthermore, the CPU 2212 may read all or necessary parts of files or databases stored on external storage media such as the hard disk drive 2224, DVD-ROM drive 2226 (DVD-ROM 2201), or IC card into the RAM 2214, and perform various types of processing on the data in the RAM 2214. The CPU 2212 then writes the processed data back to the external storage media.

[0067] Various types of information, such as various types of programs, data, tables, and databases, may be stored on the recording medium and subjected to information processing. The CPU 2212 may perform various types of processing on the data read from RAM 2214, including various types of operations, information processing, conditional judgments, conditional branching, unconditional branching, information retrieval / replacement, etc., as described throughout this disclosure and specified by the program instruction sequence, and write the results back to RAM 2214. The CPU 2212 may also retrieve information in files, databases, etc., within the recording medium. For example, if multiple entries are stored in the recording medium, each having an attribute value of a first attribute associated with an attribute value of a second attribute, the CPU 2212 may search among the multiple entries for an entry that matches the condition for which the attribute value of the first attribute is specified, read the attribute value of the second attribute stored in that entry, and thereby obtain the attribute value of the second attribute associated with the first attribute that satisfies a predetermined condition.

[0068] The programs or software modules described above may be stored on or near computer 2200 on a computer-readable medium. Alternatively, recording media such as hard disks or RAM provided within a server system connected to a dedicated communication network or the Internet can be used as computer-readable media, thereby providing programs to computer 2200 via the network.

[0069] Although the present invention has been described above using embodiments, the technical scope of the present invention is not limited to the scope described in the above embodiments. It will be apparent to those skilled in the art that various modifications or improvements can be made to the above embodiments. It will be clear from the claims that such modified or improved forms may also be included in the technical scope of the present invention.

[0070] It should be noted that the execution order of operations, procedures, steps, and stages in the devices, systems, programs, and methods shown in the claims, specifications, and drawings is not explicitly stated as "before," "prior to," etc., and that these can be performed in any order unless the output of a previous process is used in a later process. Even if the operation flow in the claims, specifications, and drawings is described using phrases such as "first," "next," etc. for convenience, this does not mean that it is mandatory to perform the operations in that order. [Explanation of Symbols]

[0071] 11 Input port, 12 Collimating lens, 13 Dichroic mirror, 14 Galvano mirror, 15 Scanning optics, 15a Incident surface, 16a Side view, 16 Lens barrel, 17 Microscope port, 18 Focusing lens, 19 Pinhole, 20 Output port, 22 Reflecting element, 23 Beam damper, 100 Scan head, 101 Laser light source, 102 Photodetector, 103 Microscope body, 104 Control unit, 105 Objective optics, 106 Sample, 107 Standby position, 108 Observable range, 200 Microscope, 2200 Computer, 2201 DVD-ROM, 2210 Host controller, 2212 CPU, 2214 RAM, 2216 Graphics controller, 2218 Display device, 2220 Input / Output controller, 2222 Communication interface, 2224 Hard disk drive, 2226 DVD-ROM drive, 2230 ROM chips, 2240 input / output chips, 2242 keyboard chips

Claims

1. A light source capable of directly controlling an ON state in which coherent light is emitted and an OFF state in which the light is not emitted, An illumination optical system that irradiates the sample with the aforementioned light to form an illuminated area, A light path changing member that changes the optical path of the aforementioned light, A control unit that controls the light source and the optical path changing member, It has, The control unit controls the optical path changing member to switch between a first state in which, when the light source is in the ON state, the light forms the illumination area on the sample, and when it is in the OFF state, the light does not form the illumination area, and a second state in which, regardless of whether the light source is in the ON or OFF state, the light does not form the illumination area on the sample. After controlling the light source to the ON state in the first state to form the illumination area on the sample with the light and terminating image acquisition, in the second state, until a command to start image acquisition is received, the light source is controlled to always be in the ON state, or controlled to alternate between the ON state and the OFF state at predetermined intervals. A microscope that, upon receiving a command to start image acquisition, controls the light source to the ON state in the first state, forms the illumination area on the sample with the light, and starts image acquisition.

2. The microscope according to claim 1, wherein the control unit controls the light source to the OFF state during at least one of the periods when switching from the first state to the second state and when switching from the second state to the first state.

3. The illumination optical system further comprises a focusing lens, The microscope according to claim 1, wherein the first state is when the light passes through the focusing lens, and the second state is when the light does not pass through the focusing lens but is incident on a predetermined standby position.

4. The device further includes a holding member for holding the light-gathering lens, The microscope according to claim 3, wherein the standby position is part of the holding member.

5. The microscope according to claim 4, wherein a light-shielding member is provided in part of the above-mentioned portion.

6. A reflective member provided at the aforementioned standby position that reflects the light, An absorbing member that absorbs at least a portion of the light reflected from the reflective member, The microscope according to claim 3, further comprising the above.

7. The microscope according to claim 1, wherein the optical path changing member is a galvanometer mirror or a spatial light modulator.

8. The microscope according to any one of claims 1 to 7, wherein the control unit, in the second state, turns the light source to the ON state, waits for a predetermined time, and then switches the light to the first state.

9. The microscope according to claim 8, wherein the predetermined time is the time it takes for the output value from the light source to stabilize.

10. A light source capable of directly controlling an ON state in which coherent light is emitted and an OFF state in which the light is not emitted, An illumination optical system that irradiates the sample with the aforementioned light to form an illuminated area, A light path changing member that changes the optical path of the aforementioned light, A control method for a microscope, comprising a control unit for controlling the light source and the optical path changing member, By controlling the optical path changing member, a first state is reached where, when the light source is in the ON state, the light forms the illumination area on the sample, and when it is in the OFF state, the illumination area is not formed. Regardless of whether the light source is in the ON state or the OFF state, it is possible to switch between a second state in which the light does not form the illumination area on the sample, After controlling the light source to the ON state in the first state to form the illumination area on the sample with the light and terminating image acquisition, the light source is controlled to remain in the ON state in the second state, or controlled to alternate between the ON state and the OFF state at predetermined intervals, until a command to start image acquisition is received. A microscope control method that, upon receiving a command to start image acquisition, controls the light source to the ON state in the first state, thereby forming the illumination area on the sample with the light and starting image acquisition.

Images