Detailed description of 2P1

  • This page documents the current state of 2P1, since Alex's 2-photon page became obsolete. To follow the step-by-step history of 2P1 see 2P1 log
  • Before planning any changes it's worth visiting the most recent version of supported parts list on Vidrio's website

Lasers

  • Coherent Chameleon Vision (A, below). Specs are here. Tunable wavelength (680-1080nm) Ti-sapphire laser, primary purpose is GCaMP imaging
  • Spark Lasers - ALCOR series 1040nm, 2W. Single wavelength laser, primary purpose is targeted optogenetic excitation

Core Microscope

  • Sutter MOM (Moveable Objective Microscope), see detailed explanation and alignment tips from Sutter
  • Secondary beam path follows the basics shown in this video, beams merged by polarising beam splitter cube (Thorlabs PBS255)

Beam paths

  • Chameleon path
    • Chameleon laser (A) - internal shutter operated from software or via panel on power supply
    • Iris - blocks reflected light from re-entering the Chameleon (B)
    • Thorlabs (WPH05M) rotating half-wave plate (C) - controls beam intensity
    • Fixed (GL10-B) Glan-Laser polariser (D) - controls beam intensity, rejected beam goes into beam trap next to it (Thorlabs BT610/M)
    • Shutter Thorlabs SH05 (E) - This shutter is normally opened by ScanImage during a run. Additionally, it is connected to an interlock on the shroud door, forced closed when the door is open.
    • Conoptics Pockels cell / EOM (F1, F2) - controls beam intensity rapidly based on input voltage (0-2V)
    • Beam sampler and photodiode, reflected portion is angle dependent (g1, g2) - used for calibrating Pockels cell, also displayed on a power meter, relationship with power after objective is linear
    • Fixed half-wave plate (H) - controls beam intensity
    • Variable beam expander (Thorlabs BE02-05-B) (I) to control objective (over/under) filling
    • Sutter MOM periscope mirror (J)
  • ALCOR path
    • ALCOR laser (a) - manual shutter on the side, normally open
    • Shutter Thorlabs SH05 (b) - operated by ScanImage trigger when this beam is operated
    • AOM from AA Opto-electronic MT110 (c ) - controls beam intensity rapidly based on input voltage (0-5V)
    • Beam sampler and photodiode, reflected portion is angle dependent (d1, d2)- used for calibrating AOM, also displayed on a power meter, relationship with power after objective is linear
    • 3x beam expander (not on picture, placed before e) - to fill objective back aperture
    • Second path periscope mirror

Scan Systems

  • Scanner for Chameleon (imaging beam): Fast scan 6mm Galvo-Galvo mirrors (Cambridge 6215) and scanner driver electronics (includes two Cambridge Technologies 671XX-HP Micromax scanner driver boards) (K)
  • Scanner for ALCOR (stimulation beam): LSKGG4/M - 4 mm Galvo-Galvo Scanner and Controller (f)
  • Scan lenses (L, g)
  • polarising beam splitter cube (M) (Thorlabs PBS255)

PMTs

Hamamatsu H7422 P40-MOD unprotected GaAsP PMTs (Hamamatsu part number, H10770PB-40 SEL) and Hamamatsu C3719 pre amps. The power supply is labeled “Sutter PS-2” but it looks suspiciously like two Hamamatsu model C3830 supplies piled into a Sutter box. I can't find any info on the PS-2, but the Hamatsu specs are here or here. This is just a guess on my part (Alex), so if you are really fussed about these specs, it would probably be best to contact Sutter.

Other Parts

  • Piezo for fast z movement (Z stacks)
    • PI725, 1CD PIFOC
    • controlled by PI E665
  • Filters/dichroic for detection
    • Chroma filter cubes housed in Olympus BX51 filter wheel
  • Table
    • T1525P; 1.5 m x 2.5 m x 210 mm Sealed Holes Metric Optical Table with PTS603 700mm High Set Of Active Legs from Thorlabs
  • Luigs-Neumann micromanipulators Mini 23 x 2, controlled by SM-5
  • Computers and Data Acquisition system
    • PC controlling the imaging system, see hardware details at 2P1 Log, hosts all data acquisition systems
      • Dev1: NI PCIe 6323:
        • Dev1/AO 0 : Stim Galvo X
        • Dev1/AO 1 : Stim Galvo Y
        • Dev1/AI 0 : Stim Galvo X position feedback
        • Dev1/AI 1 : Stim Galvo Y position feedback
        • Dev1/AO 2 : Stim AOM control
        • Dev1/AI 16: Stim power meter
        • PFIO: Trigger IN - Acquisition start
      • Dev2: NI PCI 6110 - this one's pretty old, at the next DAQ upgrade it should be changed for compatibility reasons
        • Dev2/AO 0 : Imaging Galvo X
        • Dev2/AO 1 : Imaging Galvo Y
        • Dev2 AI 0: PMT A - red
        • Dev2 AI 1: PMT B - green
      • Dev3: NI PCIe 6321
        • Dev3/AO 0 : Imaging Pockels
        • Dev3/AI 0: Imaging power meter
      • Dev4: NI PCIe 6321; Piezo input voltage and feedback
        • Dev4/AO 0: Piezo Control
        • Dev4/AI 0: Piezo position feedback
      • Dev4 could be used alternatively to be paired with MultiClamp 700B
  • Mac:
    • PointGrey Grasshopper CMOS camera controlled via MicroManager for wide-field view
    • ITC-18 DAQ board current stimuli:
      • Sync Trigger acquisition trigger sent to Dev1 PFIO (5V TTL)
      • 617nm LED controlled via Thorlabs LED driver LEDD1B with analog input (0-5V)
      • FLIR BlackFly S USB 1MP Camera recording trigger (3.3V, 15ms) - used to record behaviour with Computar MLM3X-MP 0.3-1X zoom lens
      • Custom ODD valves (5V TTL) - controller made by Electronics Workshop, Andy Howe is best to ask if they need fixing
      • Picospritzer (5V TTL)

Objectives

Software

Filters for PMTs

Alex set up the filter cube for the PMTs as below: 

         RED CHANNEL
       *-------------*
       | HQ605/75 \/ |
   *---*-------------*   
G  |   |\*
R  | E | \*
E  | T |  \*
E  | 5 |   \*
N  | 2 |    \*
   | 5 |     \*
C  | / |      \*
H  | 7 |       \*
A  | 0 |        \*
N  |   |         \*
N  |   |          \*
E  | > |           \*
L  |   |            \*
   *---*

The filters are by Chroma and the arrows point towards the coated side.


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