Virtually conjugate galvo-galvo pair
Build alignment jig
Mount galvo mirrors on shafts
Setup alignment laser
Wire up and turn on galvos
Do alignment
XXXXX Prepare V-blocks — insert photo of parts with screws
IMOHolderAssemb
1) Add
Imaging objective mounting
Assemble imaging objective mount
Gluing kinematic mount
Assembling primary dichroic cap
Putting in spring plungers and set screws
Emission splitting dichroic assembly
PMT assembly
The goal is to couple the lens onto the PMT face with oil
Seal the edge of the PMT face to the PMT housing with 5 min epoxy - careful not to soil the plate above the photocathode
Add 30 - 50 ul of high viscosity oil
Place condensor lens on oil. Make sure there are no bubbles and carefully remove overflow oil from the housing.
Place a thin ring of quick seal (WPI) on the PMT housing around the lens
Assemble alignment jigs
Alignment jig cage assembly
Imaging objective alignment jig mount
Remote focus objective assembly
The polarizing beam splitter (pbs) needs to be glued to the remote focus objective mount
The PBS needs to have the three sides marked by arrows facing the beam.
The beam needs to be flush with the alignment edges on the mount
The UV transmission of the cube is low, so be sure to expose the glue to UV for ~ 2 minutes or so
Mount the quarter wave plate as shown in the model.
Mount the remote focus objective (using 2.5 mm captive screws - Mouser 761-m0277-ss)
Left, remote focus objective mount, quarter waveplate holder, quarter waveplate, and polarizing beamsplitter. Right, components mounted on remote focus mount, including remote focus objective.
Mounting the large relays 1, 2, and 3
Relays 1 & 2 require 1.5" 1/4 20 socket screws
Relays 3 requires 2.5" 1/4 20 socket screws
Mount as shown in the photos
Left, mounts for relays 1 & 2. Right, relay 3
Assembling the covers
Assembling objetive mounted optic holder
Accessory optics
1) Remove galvo mirrors from mount
Mounting galvo driver and resonant driver boards (Spencer / Steve documentation)
Making samples
Beads
Fluorescein
Accessory optics
SECTION 2: MOUNT ON BREADBOARD
Mounting on the breadboard
Put dowel pins into holes
Mount brackets for PupilRealy3 first, make them pressed against the pins and their angled side facing down
Mount pupil relay 3, push so all the way to the left (edge against mounting surface). Tighten down clamps with screws with springs. Make sure nuts are in right place.
Mount IMOholder and tray, pushed it against the table and pins.
Mount primary dichroic, make sure caret faces down
Add primary dichro cap, pressing it against vertically mounted breadboard
Add bigDetLensHolderAssby - note need a 1/2 inch 10-24 screw to secure top right part
Add detLensCap
Mount the galvo block, pushing it against pins. Note there is not a lot of space between the galvo mirrors and pupil relay 3
Attached pupil relay 2 to the resonant chamber
Use long handle quarter-20 hex drive to mount resonant chamber to breadboard. Use paper trick to get screw sitting on ball end of hex drive, and then screw chamber down.
Attach pupil relay 1 mount to breadboard
Attach pupil relay 1
Attach remote focus objective assembly
Attach fold mirror assembly
Blow all dust out of resonant mirror chamber before sealing it up
Attach emisson splitting assembly
Attach PMT assembly, with PMT cover on. Have to thread cables through cover. Screw on filters to PMT assembly.
Attach periscope
SECTION 3: WIRING
Set up scanimage
Machine data file setting
Wiring galvos / voice coil / PMTs / etc.
SECTION 4: TABLE OPTICS ALIGNMENT
Align table optics at 900nm with the laser in alignment mode
1) Mount pockels cell just after laser, and align so that beam goes through it. Then manually set pockels cell power to about 10% for the rest of the alignment procedure.
2) Place beam expander after pockels cell. Adjust its height and the position of the front entrance such that the beam passes through the center of the iris on the beam expander. Adjust the position of the back end of the beam expander such that the beam passes straight through it. Adjust the beam expander so that the beam is collimated.
3) Position two silver mirrors such that the beam hits their centers and is steered towards the PPC. Make sure the beam remains level at its original height. Do this using an IR card stuck to a support to check beam height at different positions.
4) Place the uniblitz shutter just before the PPC in the path of the beam.
5) Adjust the positions of the enterance and exit irises at the beginning and end of the PPC so that the beam passes centeres through them.
6) Adjust height and lateral position of first two mirrors such that the beam hits both of them in the center. Then adjust the orientation of the mirrors so that the exiting beam direction is normal to entrance beam.
7) Position the large silver mirror such that the beam is centered on it, and adjust the mirror position such that the reflected beam is parallel to table and then adjust its angle it is entering prism close to its far edge.
8) Now adjust angle of prism to find the angle of minimum deviation using rotation stage (coarse). Do this by placing an IR card far away and rotating prism to find angle where beam is furthers to the right.
9) Now make sure reflected beam and incident beam are coplanar by adjusting the angle of the prism and looking at the height of the reflected beam on the IR card. The reflection is very faint and needs an IR viewer to be seen.
10) Now make sure the refracted beam is coplanar with the reflected and incident beam by adjusting the other angle of the prism.
11) Redo finding angle of minimum deviation using fine adjustment. This will be the angle where the beam is the rightmost possible.
12) Algin retroreflector axis. Adjust height of retreflector such that the return beam is centered on the roof mirror. Then adjust close side of rail such that the returning beam is hitting the prism a little further into the prism (2 to 3 mm). Then adjust the back position such the rail is exactly parallel to the beam. Slide the retroreflector back and forth along the rail to check that it is moving along the axis of the beam.
13) Adjust lateral position of roof prisms so that return beam is entering the prism, and rotate it so that the outbound beam 3 is hitting the retroreflector. Do fine angle adjustments of the prisms angle such that return beam 4 is level to table and colinear with track.
14) Mount output coupler. Place in bypass mode, and set target card after exit iris. Left bypass mode. Align exit beam to pass through iris and hit target by adjusting height of retroreflector so that beam passes through center of exit iris and move mirror to translate mirror, and use angle of mirror to make beam hit target on iris card.
15) Add two mirrors such that beam is level as it goes into the entrance of periscope.
16) Set periscope entrance mirror center to be 120 mm heigh (height of laser beam).
Pockels cell
Shutter
2x table beam expander
Assembling the compressor
Build enclosure for table optics
SECTION 5: MICROSCOPE ALIGNMENT
Multistage periscope alignment
Quarter wave plate adjustment for multistage periscope
Alignment of beam to remote focus objective
Adjustment of quarter wave plates for maximum transmission
Alignment of RF mirror assembly
Alignment of beam to imaging objective using compensators
Adjustment of galvo voltage divider gain
SECTION 6: IMAGING
Dye pool imaging to set compensators
Dye pool imaging to set retroreflector position
Image beads and measure PSF
APPENDIX A:
Accessory optical path
Assembly, alignment, calibration, adjustment
General description: The accessory optical systems of the 2p-RAM couple through a 52 x 72 x 1 mm dichroic/mirror that is placed in the 2p-RAM detection path. This optic can be moved in and out of the optical path with a motor. Its size does NOT support the full FOV and NA of the 2p-RAM imaging or detection paths, but the full FOV is supported at a reduced NA. All light to/from the accessory optical systems goes through the 2p-RAM primary dichroic, so must have a wavelength shorter than the prim dichro cutoff (currently ~720nm) for efficient transmission. The main 2p-RAM tube lens is not part of the accessory optical systems path, so an accessory tube lens is used. All accessory optical systems are light-tight, so little ambient light will enter the 2p-RAM detection path.
The following functionality is implemented:
- Scanning 1p photostimulation system (S1PS): The S1PS roughly images the face of an SMA-terminated optical fiber onto the sample, with a magnification of 1.85x. It supports a fiber NA of up to 0.22 without vignetting. A galvo-galvo scan mirror pair dedicated to this system can scan the fiber-face image anywhere within the 2p-RAM 5mm FOV. It is intended to be used with multimode fibers, such that the spot size at the sample can be changed by changing the fiber core diameter. No light source or fiber is provided. A manual, quick-change filter mount holds a dichroic mirror that couples the S1PS system with the wide-field imaging system, which can be used simultaneously, if their wavelengths are separable.
- Wide-field imaging system (WFIS): The WFIS images the sample onto a CCD camera, with epi-illumination. A quick-change filter cube couples the epi-illumination and imaging, allowing switching between epi-fluorescence imaging and polarization- or absorption-based imaging using a polarizing/non-polarizing beamsplitter cube (some scattering off of this cube from the epi-illumination is visible in the image obtained in the non-fluorescence mode). The epi-illumination module accepts light from a 3mm core diameter liquid light-guide (LLG face is imaged onto objective pupil; NA .24 accepted from LLG; field stop is provided). The imaging goes through a 35mm fixed focal length imaging lens, providing an aperture stop. As configured, FOV at the sample is 4.8 x 6mm, magnification is 1.13x onto the camera, and resolution should be limited by pixel size (5.3 um; 1024 x 1280).
- Coverglass alignment system (CAS): The CAS provides real-time measurement of the angle of the sample coverglass relative to the 2p-RAM objective lens. A 100um pinhole is illuminated by an LED, and then imaged onto the 2p-RAM objective pupil, producing a nearly collimated beam at the sample (beam width is controlled by a variable iris placed next to LED). If no immersion water is used, then the reflected beam from the top coverglass surface is focused onto a camera through a 50:50 beamsplitter. The spot position on the camera gives the coverglass angle, with resolution exceeding 0.1 degree. The CAS is coupled to the S1PS and WFIS with a motorized flip-mirror. The CAS needs to be calibrated for zero-angle. The design for a jig for this calibration, which precisely holds a piece of glass relative to the 2p-RAM objective, will be provided.