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docs:manual:assembly [2015/06/25 22:31] Jon Daniels created |
docs:manual:assembly [2015/06/26 18:29] Jon Daniels [With 2015 piezos] |
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Next remove the piezo objective mover (APZOBJ) from the SPIM arm mount as shown in <imgref piezo_screws_close> | Next remove the piezo objective mover (APZOBJ) from the SPIM arm mount as shown in <imgref piezo_screws_close> | ||
+ | If needed, now is the time to insert the objective on the other side of the diSPIM. It is easy to access the side of the diSPIM that was left intact now that much of the other side is removed (if needed you can raise the upper Z stage). | ||
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+ | Re-attach the piezo objective mover with objectives by screwing it into the SPIM arm mounting plate. Finally, reattach the SPIM arm to the arm mount. | ||
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+ | With everything reassembled, | ||
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+ | We recommend avoiding use of the vertical objective position screw (<imgref LowerAdj> | ||
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+ | < | ||
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+ | In further adjustments we will move the objective focus bushings on both sides to focus and manipulate the right objective laterally using the lateral objective screw (<imgref LateralAdj> | ||
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+ | ==== With 2015 piezos ==== | ||
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+ | Inserting the objectives is significantly easier with the 2015 piezo mounting scheme. The piezo actuators and everything attached to them slide in and out of the SPIM arm mount (RAO-0046 ) using a dovetail mechanism. A new lateral fine adjuster sits between the piezo objective mover and the SPIM arm mount. The objectives are brought into co-focus in 3-dimensional space by three orthogonal adjustments: | ||
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+ | To remove the piezos from the SPIM arm mount, first loosen the set screw on the bottom of the arm mount; it is the center screw of the three on the side of the SPIM arm mount as shown in <imgref piezo_dovetail> | ||
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+ | Upon re-insertion, | ||
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+ | With the objectives inserted and everything reassembled, | ||
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+ | ===== Care of piezo objective movers ===== | ||
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+ | The piezo objective movers are the most failure-prone component of the diSPIM system. It appears that the piezo actuators can be damaged by external stresses, including as screwing the objective bushings in so far that the piezo top plate is moved mechanically (impossible with 2015 piezos or more recent; for older systems be sure to follow the instructions in Section [sub: | ||
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+ | During normal use the piezos are also stressed electronically. Because the electronic stress scales quadratically with applied voltage, being in the extreme negative position (near the sample) wears out the piezo faster than using the piezo near the center position. Of course electronic stresses are part of normal operation, but if the diSPIM is not being used for a long period of time then we recommend either turning off the Tiger controller or else using the command to disable the piezo axes (this reduces the applied voltage to even less than the center position). In Micro-Manager this can be done easily using the “MotorOnOff” property of the piezo stages, setting its value to be “On” in the System-Startup configuration preset and “Off” in the System-Shutdown preset (see the [[https:// | ||
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+ | As of December 2014 we recommend 150 um travel piezo objective movers with increased flexure width, which reduces the tendency for side-to-side vibration of the piezo top plate in response to other vibrations (e.g. from the camera fans or imperfect isolation from the floor). | ||
+ | |||
+ | ===== Tiger Controller ===== | ||
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+ | The motion components of the diSPIM microscope are controlled by ASI’s modular Tiger controller. The controller should be located near the microscope such that all cables reach and so that the indicator lights on the face of the control units can be seen easily. | ||
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+ | < | ||
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+ | A typical diSPIM controller contains the following modules: | ||
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+ | ==== TG/COM ==== | ||
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+ | Communications card with USB connection to host computer. Supports four-axis joystick/ | ||
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+ | ==== X/Y ==== | ||
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+ | Two-axis XY stage card. Also supports a dimmable LED illuminator. | ||
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+ | ==== Z/F ==== | ||
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+ | Two-axis card for SPIM focus (F) and lower microscope focus control (Z). May also have an LED illuminator. | ||
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+ | ==== MICRO-MIRROR ==== | ||
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+ | Four-axis micro-mirror controller card. Controls two standard light sheet scanners, or a single scanner with anti-striping micromirrors. Control logic for synchronizing the light sheet, camera triggers, and piezo motion resides on processor in this unit. | ||
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+ | ==== TTL / PLC ==== | ||
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+ | TTL buffer card or Programmable Logic card outputs lasers and camera control signals. | ||
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+ | ==== PIEZO P ==== | ||
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+ | P-axis objective piezo positioner card. | ||
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+ | ==== PIEZO Q ==== | ||
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+ | Q-axis objective piezo positioner card. | ||
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+ | ===== Connecting cables ===== | ||
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+ | Cables to all components are labelled. On the diSPIM there are identical cameras scanners and piezos on both sides so it is important to keep straight which cables go where so that the control software can work as intended. | ||
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+ | ==== Tiger Controller ==== | ||
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+ | As a safety precaution the Tiger controller should always be powered off when connecting or disconnecting cables. | ||
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+ | Install all motion control cables to the appropriate Tiger controller module. Be careful that X/Y and Z/F cables are not interchanged. Check serial numbers on the piezo objective movers and controller face-plates to be sure to connect the cables to the correct controller module. | ||
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+ | Connect the Tiger USB connection to the host computer. If the USB to serial driver doesn’t install automatically, | ||
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+ | ==== Programmable Logic Card (or TTL) Card Connections ==== | ||
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+ | The Programmable Logic Card, or PLC, was introduced in 2015 and is used for the TTL control of the cameras and lasers. Its outputs are generally | ||
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+ | - Path A camera trigger (right-side camera) | ||
+ | - Path B camera trigger (left-side camera) | ||
+ | - Not connected (can be configured as global shutter or ongoing acquisition signal) | ||
+ | - Laser side select | ||
+ | - Laser #1 on/off | ||
+ | - Laser #2 on/off | ||
+ | - Laser #3 on/off | ||
+ | - Laser #4 on/off | ||
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+ | Which lasers are #1–4 does not matter, because the mapping between PLC output and a user-defined label is specified in software. In Micro-Manager, | ||
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+ | If you have a TTL card (non-upgraded systems shipped in 2014 or earlier), CAM0 and CAM1 are for camera triggers of paths A and B respectively. LSR0 is for the laser on/off control, and LSR1 corresponds to the laser side select. LSR0 and LSR1 may be connected differently if you have a non-standard laser configuration; | ||
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+ | ==== Light Path A Component Connections ==== | ||
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+ | * Left-side scanner is connected to cable end marked **BA**.((Before March 2014 other cables and scanner orientations were used. Contact ASI for new cables.)) | ||
+ | * Right-side imaging piezo P is connected to the P-axis Piezo card. | ||
+ | * Right side camera Trigger is connected to PLC #1 (CAM 0 on the TTL card). | ||
+ | |||
+ | ==== Light Path B Component Connections ==== | ||
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+ | * Right-side scanner is connected to the cable end marked **DC**. | ||
+ | * Left-side imaging piezo Q is connected to the Q-axis Piezo card. | ||
+ | * Left-side camera Trigger is connected to PLC #2 (CAM 1 on the TTL card). | ||
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+ | ==== Cameras ==== | ||
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+ | Install the camera cards in the host computer according to the manufacturer’s instructions, | ||
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+ | Connect the camera data cables to the camera cards in the computer. The camera trigger cables should be connected as described. | ||
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+ | ===== Laser and Fiber Installation ===== | ||
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+ | There are many possible laser configurations that are theoretically supported. A bare-bones approach uses a single fiber coupled laser which is split with a fiber splitter into two outputs for the two scanners. In this configuration both output fibers have light simultaneously. The scanners act as imperfect shutters (~0.1% transmission) when steered to their blanking position. | ||
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+ | More commonly, users have a laser merge module with dual outputs which controls of multiple laser lines and includes a routing switch that will direct the laser output one of two output fibers. The outputs on the TTL or PLC cards control the lasers. For a single-color setup, connect the TTL laser on/off control to LSR 0 and the fiber-switching signal to LSR 1 on the TTL card. For control of multiple laser lines, the PLC card is required. Generally PLC outputs #5-#8 are connected to the respective laser on/off controls and #4 is connected to the fiber-switching input. | ||
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+ | ===== Install filters, Filter cubes and Mirrors ===== | ||
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+ | Dichroic mirrors, emission and excitation filters should be installed in the C60-D-CUBE for each SPIM arm. Similarly, install the right angle mirrors for the camera tubes in the appropriate cubes. Details are shown in <imgref D-Cube>. | ||
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+ | Remove the two thumbscrews the hold the front dovetail mount, C60-DOVE-II , in place. Give a slight tug and twist so the magnets holding the part in place will release the cover assembly from the cube body. Remove the dovetail mount section from the MIM-CUBE-II’s covers as shown in <imgref Dove-II> | ||
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+ | ==== How to adjust the mirror cubes ==== | ||
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+ | The kinematic adjusters are used during alignment to tilt the mirrors in the MIM-CUBE-II ’s, both for camera mirrors and dichroic mirrors. You will have best results if you follow these steps. | ||
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+ | - Loosen the thumb screws several turns. **Do not manipulate the adjustment screws while the thumb screws are engaged; doing so can strip the adjustment screws.** | ||
+ | - Grasp the cube body a apply modest pressure to the center of the adjustable face with your thumb to firmly push the kinematic adjusters into their seats. See Figure [fig: | ||
+ | - Turn the three adjustment screws as necessary to steer the mirror using a 3/32” Allen driver. | ||
+ | - When where desired, lightly snug down the thumb screws and only then release your pressure on the cube face. Slight movement may occur depending on the order and tightness in which you tighten the thumb screws, which can be taken advantage of to make tiny adjustments. |