Sofroniew, Flickinger et al eLife 2016

CAD Models 

You can view the files in the Autodesk free viewer https://a360.autodesk.com/viewer/

Understanding and interpreting the models is critical for building. 

2p-RAM full model with gantry and accessory optics

2p-RAM_20160725.dwfx

2p-RAM without gantry, with accessory optics

2p-RAM without gantry and accessory optics

2p-RAM_NoGantryNoAccessoryOptics_20160725.dwfx

Prism pulse compressor – on optical table

PPCompressor_Version201607.dwfx

Mounting the resonant mirror - including alignment jig

ResMirrAlignmentAssembly.dwfx

GalvoAlignmentAssby.dwfx

2p-RAM main optical path

Preprint

Accessory path

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: 

1. 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. 
2. 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).
3. 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.