Multiphoton Resources

1. Zeiss LSM 510 META NLO Two-Photon Laser Scanning Confocal Microscope System

Coupled with:

  • Zeiss Inverted Axiovert 200 M Microscope
  • Zeiss CTI Controller 3700
  • Zeiss Tempcontrol 37-2 Digital

(1) Laser Lines

  • Argon: 458/477/488/514 nm, 25mW
  • HeNe1: 543 nm, 1mW
  • HeNe2: 633 nm, 5mW
  • Chameleon XR Laser System: tunable, 705 nm – 980 nm

Chameleon laser excites fluorophores from Blue to Red range, but most efficient with UV and near-UV excitable dyes.

(2) Objectives

Objectives available for Zeiss Axiovert 200 M inverted microscope:

Mag. Objective NA Medium WD(mm) For 2-Photon
10x Plan-Apochromat 0.45 Air 2.10 No
20x Plan-Apochromat 0.75 Air 0.61 Yes
40x Plan-Neofluar 1.30 Oil (DIC) 0.17 Yes
63x C-Apochromat 1.20 Water (Corr.) 0.24 No
100x Alpha Plan-Fluar 1.45 Oil 0.10 No

 

(3) Fluorescence Filter Sets

Fluorescence filter sets available for use with the Zeiss Axiovert 200 M inverted microscope:

  • CFP filter set
  • FITC filter set
  • Rhodamine filter set
  • YFP filter set
  • DAPI filter set
  • DAPI/ FITC/Texas Red filter set

 

2. Olympus BX61WI InVivo Two-Photon Confocal Microscope

(1) Laser Line

Coherent Chameleon XR Laser System: tunable, 705 nm – 980 nm

(2) Objectives

Objectives available for Olympus BX61WI upright microscope:

Mag. Objective NA Medium WD(mm) For 2-Photon
10x UMPLFL 0.30 Water 3.30 No
60x LUMPLFL (IR2) 0.90 Water 2.00 Yes

(3) Filter Sets

Fluorescence filter sets available for Two-Photon imaging:

  • GFP/DsRed Filter Set (Dichroic mirror: T560LPXR):

    Channel 1: GFP (ET510 - 80m 2p) –Emission filter: 470-550 nm

    Channel 2: dsRed (ET620 - 60m 2p) –Emission filter: 590-650 nm

  • CFP/YFP Filter Set (Dichroic mirror: T505LP):

    Channel 1: CFP (ET480 - 40m 2p) –Emission filter: 460-500 nm

    Channel 2: YFP (ET535 - 30m 2p) –Emission filter: 520-550 nm

3. Nikon N-SIM Super Resolution Microscope System

(http://www.nikon.com/products/instruments/lineup/bioscience/s-resolution/nsim/index.htm)

N-SIM can produce nearly two times the resolution of conventional optical microscopes by combining “Structured Illumination Microscopy” (SIM) technology and Nikon Eclipse Ti research inverted microscope, with lateral resolution to approximately 100nm and axial resolution to approximately 200nm. 3D capable. TIRF capable. Laser lines: 405nm, 488nm, 561nm, 640nm.

4. Nikon N-STORM Super Resolution Microscope System

(http://www.nikon.com/products/instruments/lineup/bioscience/s-resolution/nstorm/index.htm)

N-STORM can produce 10 times the resolution of conventional optical microscopes by combining “Stochastic Optical Reconstruction Microscopy” (STORM) technology and Nikon's Eclipse Ti research inverted microscope, with lateral resolution to approximately 20nm and axial resolution to approximately 50nm, extending the role of the optical microscope to near molecular level resolution. 3D capable. Laser lines: 405nm, 488nm, 561nm, 647nm.
          STORM Protocol-Sample Preparation
(https://content.ilabsolutions.com/wp-content/uploads/2014/06/N-STORM-PROTOCOL.pdf)

5. Image Processing Software

One standalone workstation with Nikon NIS-Element (Ar) software, STORM images, and SIM images off-line analysis licenses.
One standalone workstation with Zeiss LSM software (version 4.0) off-line image analysis license

6. Preparative Equipment

(1) Cell Culture Room
• Class II Biological Safe Cabinet, Baker Company
• CO2 Incubator, SANYO
• 4 C° Refrigerator, SANYO