Two-photon excitation laser-scanning microscope

The LMCF two-photon microscope is in beta form and ready for use. The system is physically complete but we are waiting for LSMtech to complete the new software (currently averaging is unstable, and it lacks the capability to acquire z-stacks).

Duke University Light Microscopy Core Facility 2 photon microscope Capabilities:
  • Multi-photon excitation
  • Imaging thick samples
  • Multi-wavelength
  • Standard 1 photon imaging

Location: Room 4228 French Family Science Center (613-8097)

Cost: $40 per hour

User manual:

Request training | Reserve instrument

Detailed specifications of the system (further information)

The system is based on a Zeiss 410 confocal.

MPE laser:

Detectors:
  • Two channel non-descanned external detector (green/red, can add other filters)
  • Three descanned PMTs (see emission filters below)

Objectives available:

  • 10x/0.50 (Zeiss Fluar 44 01 33)
  • 20x/0.50 Ph2 (Zeiss Plan-NeoFluar 44 03 41)
  • 25x/0.80 multi-immersion (Zeiss Plan-NeoFluar 44 05 42)
  • 40x/0.8 W Achroplan dipping objective (44 00 90)
  • 40x/1.30 oil DIC (Zeiss Plan Neo-Fluar 1056-602)
  • 63x/0.95 W dipping objective (44 00 67)
  • 63x/1.40 oil (Plan Apochromat 44 07 60)
  • 100x/1.30 oil (Zeiss Plan Neo-Fluar 44 04 80)
  • Others as requested

Microscope and stage:

Computer:

  • Custom-built Windows XP computer
  • LabView based LSM controller
  • GUI for laser and pre-comp control
  • Original Zeiss software and computer available in parallel (used for maintenance)

 

Conventional single photon imaging:

Lasers:
  • Omnichrome KrAr (488, 568 nm)
  • HeNe (633 nm)

Dichroics:

  • DBS1 Two-photon, NT 80/20, UV/488/568/633 quad
  • DBS2 LP560, LP630, open
  • DBS3 open, LP460, LP560

PMTs:

  • R/FL1 - cooled
  • R/FL2 - cooled
  • R/FL3 - standard

Emission Filters: Dichroic/filter combinations: gif pdf

PMT1 emission filters

  • LP 400 (DAPI)
  • BP 470-500 (INDO)
  • LP 505 (GFP, FITC, Alexa 488)
  • LP 510-560 (GFP, FITC, Alexa 488)
  • LP 590 (CY3/Rhod/TxRd)
  • BP 575-635 (CY3/Rhod/TxRd)
  • BP 615-645 (TxRed)
  • BP 670-810 (CY5)

PMT2 emission filters

  • BP 400-440 (DAPI)
  • BP 390-420 (INDO)
  • BP 500-530 (GFP, FITC, Alexa 488)
  • BP 500-540 (GFP, FITC, Alexa 488)
  • BP 510-560 (GFP, FITC, Alexa 488)
  • BP 580-620 (Rhod/CY3/TxRd)
  • BP 575-650 (CY3/TxRd)

PMT3 emission filters

  • LP 400 (DAPI)
  • BP 400-440 (DAPI)
  • BP 460-500 (INDO)
  • BP 500-530 (GFP, FITC, Alexa 488)
  • BP 500-540 (GFP, FITC, Alexa 488)
  • BP 510-560 (GFP, FITC, Alexa 488)
  • BP 575-650 (CY3/TxRd)