Confocals

Nikon Ti2E AXR NSPARC (Porter B049)

Lasers & Filters

Lasers:

Launch: LUD-S4Ìý

Excitation at: 405, 488, 561, 640 (nm)

Emission (Chroma) and molecules imaged:

  • Emission filter: 425-475 (nm): DAPI, Hoechst, DyLight-405, Alexa405, etc.
  • Emission filter: 500-550 (nm): Alexa488, FITC, EGFP, Opal520, 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)

Light source and filters for viewing by eye:

• Nikon D-LEDI; 385, 475, 550, 621 nm for DAPI, GFP, RFP, Cy5 with multi-band emitter, brightfield

NSPARC super-resolution lens:

  • 60X PLAN APO TIRF oil NA 1.49 WD 0.12 (mm), DIC

Additional standard objectives:

  • 2x PLAN APO Ìýλ NA 0.10, WD 8500 (mm)
  • 10X PLAN APO λ D NA 0.45 WD 4.0 (mm), DIC
  • 20X CFI60 PLAN APO λ D ÌýNA 0.80 WD 0.80 (mm), DIC
  • 40XC PLAN APO λ S ÌýSILICONE NA 1.25 WD 0.30 (mm), DIC, correction collar (23-37 degrees C)
  • 60X PLAN APO IR SR water 1.27NA WD 0.18 (mm), DIC

Available upon request:

  • 60x 1.40NA Plan Apo OIL VC λ WD 0.13 (mm)
  • 40x NA 0.95 WD 0.18 (mm)
  • inquire
  • DUX-VB, 4 PMT-GA GaAsP
  • Nikon Spatial Array Confocal (NSPARC) pinhole array sensor
  • High frame-rate imaging using resonance scanner
  • Super resolution to 100nm XY using NSPARC detector
  • Adjustable GFP and RFP emission bandwidth (confocal only)
  • TOKAI-HIT Cell Incubator 1 x 35 (mm) dish, chamber side (coverslip bottom), multiwell plate
  • Motorized X, Y, Z and separate Prior QGNPS083-NI Piezo 600 micron Z-stage
  • Multi-point, time-lapse, optical sectioning, large-area montage with stitching
  • Advanced analysis tools via separate HP Z4 workstation, including spectral deconvolution, and full analysis suite (inquire with Director)
  • NIS Elements AR 6.02.03 with JOBS, general analysis
AX NSPARC front view

in progress

THOR - Nikon TiE A1 laser scanning, structured illumination and TIRF (Porter B047A)

Lasers & Filters

Lasers:

Launch: LUNV6, high-output

Excitation at: 405, 445, 488, 514, 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 super-resolution lens:

  • 100x 1.49NA OIL SR Apo TIRF

Additional standard objectives:

  • 4x 0.2NA Plan ApoÌýλ WD 20 (mm)
  • 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 1.27NA Water SR Plan Apo WD 0.17 (mm)

Available upon request:Ìý

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

Laser Scanning Confocal:

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

N-SIM:

  • Andor iXON 897 Ultra EMCCD (512x512)

Ìý

Ìý

Ìý

  • 3D structured illumination, 'fast' 2D SIM
  • TOKAI-HIT Cell Incubator 1 x 35 (mm) dish, chamber side (coverslip bottom), multiwell plate
  • Motorized X, Y, Z and separate MCL Piezo Z with 200 micron range
  • Multi-point, time-lapse, optical sectioning, large-area montage with stitching
  • Advanced analysis tools via separate workstation, including spectral deconvoltion, and full analysis suite (inquire with Director)
  • ÌýAR 5.04; 64 bit
Thor
PSF of 100 nm tetraspeck bead
  • Please contact Director for training

BLACK WIDOW - Nikon TiE Eclipse spinning disk with FRAP (Porter B049)

Lasers & Filters

Lasers

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

Emission (Single bandwidth are Chroma and molecules imaged:

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

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:Ìý

  • 4xÌý0.13NA Plan Fluor WD 17.2 (mm)
  • 60xÌý1.27NA 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 nmÌýlaser (50mW)
  • Motorized X, Y, Z
  • TOKAI-HIT Cell Incubator; 1 X 35 (mm) dish or chamber slide (coverslip bottom).
Black Widow

CHRONOS - Yokogawa CellVoyagerâ„¢ CV1000 spinning disk scanner (Porter B047A)

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

Standard objectives: All lenses are Olympus

  • 10x 0.4NA UPLSAPO WD 3.1 (mm)
  • 20x 0.75NA UPLSAPO WD 0.65 (mm)
  • 20xÌý0.45NAÌýLUCPLFLN 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 13Ìýmicron 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

ChronosÌýis equipped with an ultra-sensitive, high-resolutionÌýmode Hamamatsu EMCCD cameraÌýand a Yokogawa X1 spinning disk with the option ofÌýdual microlens-enhanced Nipkow scanning disks with pinholes ofÌý50 μm or 25 μm; disks can also be removed for widefield imaging. Zero Drift (Olympus) autofocusing and software focusing.ÌýThis microscope has minimalÌýphototoxicity and photobleaching, andÌýis 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 wellsÌý(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.ÌýPlease 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
Chronos, front