Confocals

THOR - Nikon A1 laser scanning, structured illumination and TIRF

Lasers & Filters

Lasers:

Lauch: LUNV6, high-output.

Excitation at: 405, 445, 488, 515, 561, 647 (nm)

Emission (Chroma) and molecules imaged:

  • Emission filter: 425-475 (nm): DAPI, Hoechst, DyLight-405, Alexa405, etc.
  • Emission filter: 467.5-502.5 (nm): CFP, cerulean, etc.
  • Emission filter: 500-550 (nm): Alexa488, FITC, EGFP, Opal520, etc.
  • Emission filter: 521.5-554.5 (nm): YFP, Venus, etc.
  • Emission filter: 575-625 (nm): Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, Opal570, etc.
  • Emission filter: 650-720 (nm): Alexa647, Cy5, Opal620, etc.
  • DIC imaging possible (laser scanning confocal only)

Filters for widefield:

DAPI, GFP, RFP, Brightfield

N-SIM certified objectives:

  • 100x 1.49NA OIL SR Apo TIRF WD 0.12 (mm)

  • 60X 1.27NA Water SR Plan Apo WD 0.17 (mm)

Standard objectives:

  • 4x 0.2NA Plan Apoλ WD 20 (mm)
  • 10x 0.45NA Plan Apo λ WD 4 (mm)
  • 20x 0.75NA Plan Apo VC λ WD 1 (mm)
  • 40x 0.6NA S Plan Fluor ELWD 3.6-2.8 (mm)
  • 40x 0.95NA Plan Apo λ WD 0.25-0.17 (mm)

Available upon request:

  • 60x 1.40NA Plan Apo OIL VC λ WD 0.13 (mm)

Laser Scanning Confocal:

  • 2 x GaAsP high-sensitivty PMTs (CFP, GFP, YFP, RFP)
  • 2 x multi-alkali PMTs (DAPI, far-red)

N-SIM:

512 X 512 16 bit

  • TOKAI-HIT Cell Incubator 1 x 35 (mm) dish or chamber side (coverslip bottom)
  • Motorized X, Y, Z and separate Mad City Labs Piezo Z
  • Multi-point, time-lapse, optical sectioning, large-area montage with stitching
  • Increase power over imaging depth
  • 5.21.03; 64 bit

THOR - Nikon Structured Illumination Super-resolution and A1 Laser Scanning Confocal Microscope (Left View)

THOR - Nikon Structured Illumination Super-resolution and A1 Laser Scanning Confocal Microscope (Front View)

THOR - Nikon Structured Illumination Super-resolution and A1 Laser Scanning Confocal Microscope (Right View)

THOR - Raw Data of 3-D SIM Image Showing 5 Phase Translations Per Grid Rotation and 3 Rotational Angles.

THOR - Nikon Raw Data of 3-D SIM Image Showing Interference Fringes (Images provided by the Orth Lab).

BLACK WIDOW - Nikon Ti Eclipse spinning disk with FRAP

Lasers & Filters

Lasers

  • 405, 488, 561, 640(nm)
  • Quad used for excitation transmission in photobleaching or photoactivation experimentsis Semrock Brightline Di01-T405/488/568/647

Emission (Single bandwidth are Chroma and molecules imaged:

  • Emission filter: 430-480(nm): DAPI, Hoechst, DyLight-405, Alexa405, BFP, etc.
  • Emission filter: 500-540 (nm): Alexa488, FITC, EGFP, Opal520, etc.
  • Emission filter: 602-662(nm): Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, Opal570, etc.
  • Emission filter: 669-741(nm): Alexa647, Cy5, Opal620, etc.

Standard objectives:

  • 10x 0.45NA Plan Apo λ WD 4 (mm)
  • 20x 0.75NA Plan Apo VC λ WD 1 (mm)
  • 40x 0.95NA Plan Apo λ WD 0.25-0.17 (mm)
  • 60x 1.40NA Plan Apo VC λ OIL WD 0.13 (mm)
  • 100x 1.45NA Plan Apo λ OIL WD 0.13 (mm)

    Available upon request:

    • 4x0.13NA Plan Fluor WD 17.2 (mm)
    • 60x1.27NA water SR Plan Apo WD 0.17 (mm)
    • Hamamatsu ImagEM EM-CCD (widefield)
    • Both cameras are equipped with 512 X 512 16-bit imaging array
    • Fast Acquisition
    • Multipoint, montaging, stiching
    • Single point FRAP/PA with 405 nmlaser (50mW)
    • Motorized X, Y, Z
    • TOKAI-HIT Cell Incubator; 1 X 35 (mm) dish or chamber slide (coverslip bottom).

    “Live cell imaging of a sarcoma cell expressing lamin A-EGFP and subsequent photo-bleaching event. Note the photobleached spot that appears in the lamin A on the left, near the end of the movie. 100X NA1.45 oil and 405 nm laser was used for photobleaching.”

    BLACK WIDOW - Nikon Inverted Spinning Disk Confocal (Right View)

    BLACK WIDOW - Nikon Inverted Spinning Disk Confocal (Laser On)

    BLACK WIDOW - Nikon Inverted Spinning Disk Confocal (Right View)

    Chronos - Yokogawa & Olympus CellVoyager™ CV1000 spinning disk scanner

    Lasers & Filters

    Lasers

    • 405, 488, 561 (nm) & Brightfield via far-red LED

    Emission filters are from Olympus and molecules imaged:

    • Emission filter: 447-507 (nm): DAPI, Hoechst, BFP, etc..
    • CFP has been imaged using the 405 nm laser with 447-507 (nm) emission filter.
    • Emission filter: 500-550(nm): Alexa488, FITC, EGFP, etc.
    • Venus has been imaged using the 488 nm laser with 500-550 (nm) emission filter.
    • Emission filter: 580-654(nm): Alexa568, 594, TxRed, TRITC, mRFP, mCherry, mKO, TdTomato. etc.

    Standard objectives:

    • 10x 0.4NA UPLSAPO WD 3.1 (mm)
    • 20x 0.75NA UPLSAPO WD 0.65 (mm)
    • 20x0.45NALUCPLFLN LWD WD 6.4-7.6 (mm)
    • 40x 0.6NA LUCPLFLN LWD WD 2.7-4.0 (mm)
    • 40x 1.30NA UPLFLN OIL WD 0.2 (mm)
    • 100x 1.40NA UPLSAPO OIL WD 0.13 (mm)

    Objectives available upon request:

    • 4x 0.13NA UPlanFl N UIS2 WD 17 (mm)
    • 60x 1.20NA UPLSAPO WATER WD 0.28 (mm)
    • 60x 1.35NA UPLSAPO OIL WD 0.15 (mm)

    • This is a 1024 x 1024, 16 bit detector with 13 x 13micron pixels
    • Back thinned FT CCD with >90% QE from 500-700 (nm)
    • Cooled to -55 degrees C to reduce detector dark noise
    • Camera binning 2x2, 4x4 to increase sensitivity

    The nitty gritty

    Chronosis equipped with an ultra-sensitive, high-resolutionmode Hamamatsu EMCCD cameraand a Yokogawa X1 spinning disk with the option ofdual microlens-enhanced Nipkow scanning disks with pinholes of50 μm or 25 μm; disks can also be removed for widefield imaging. Zero Drift (Olympus) autofocusing and software focusing.This microscope has minimalphototoxicity and photobleaching, andis therefore great for live cell imaging at high temporal resolution, multi-point, and multi-day time-lapse. Multi-point montaging of large tissue samples in X-Y-Z and low-to-medium throughput screening. Excellent brightfield imaging through the disk. Can reload previous sample map and imaging positions to re-image the same locations at different magnifications or over time.

    Standard sample holders

    • Standard slides
    • Multiwell slides
    • Multiwell plates, including 6, 24, 96 and 384 wells(please inquire, there are some plate restrictions for live-imaging)

    Environmentally controlled chambers

    • Single 35 (mm) dish
    • Triple 35 (mm) dish
    • Multiwell plates including 6, 24, 96 and 384 wells. Please note: this incubation chamber can only accept low profile plates, standard profile do not fit into the chamber.Please inquire.

    Acquisition

    • CV1000 software 1.06.06

    Viewer

    • CV1000 viewer software version 1.06.06 available for users (please email Director or Specialist for copy)
    • ImageJ

    Kronos - Yokogawa CellVoyager™ CV1000 Confocal Scanner System (Front View Stage Out)

    Kronos - Yokogawa CellVoyager™ CV1000 Confocal Scanner System (Side View)

    Kronos - Yokogawa CellVoyager™ CV1000 Confocal Scanner System (Environmental Controls)

    IRON MAN - Zeiss 510 laser scanning confocal - OUT OF SERVICE

    Lasers & Filters

    Lasers

    • Argon/2 - 458, 477, 488, 514 (nm)
    • HeNe - 543, 633 (nm)
    • Diode - 405/30 (nm)

    Molecules imaged:

    • DAPI, Hoechst, similar
    • CFP
    • Alexa488, FITC, EGFP
    • Alexa568, 594, TxRed, TRITC, mRFP, mKO
    • Alexa647, Cy5

    Filters

    • DAPI, EGFP, RFP, Brightfield

    Standard objectives

    • 5x 0.15NA EC Plan-Neofluar WD 3.1 (mm)
    • 10x 0.30NA EC Plan-Neofluar WD 4.0 (mm)
    • 20X 0.80NA Plan Apochromat WD 0.5 (mm)
    • 40x 1.30NA EC Plan-Neofluar OIL WD 0.21 (mm)
    • 63x 1.40NA Plan Apochromat OIL WD 0.19 (mm)
    • 100x 1.40NA Plan Apochromat OIL WD 0.17 (mm)
    • 3x PMT, standard
    • Multichannel
    • FRET
    • FRAP
    • Z-Drive
    • 2009

    Iron Man - Zeiss 510 Confocal Laser Scanning Microscope (Front View)

    Iron Man - Zeiss 510 Confocal Laser Scanning Microscope (Left View)