Confocals

THOR - Nikon A1 laser scanning, structured illumination and TIRF

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 Apo λ 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:

Andor iXon X3 EM-CCD 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
SOLA Light engine
Nikon Perfect Focus System

NIS Elements 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 - Nikon Super-resolution (SIM) - Raw Data of 3-D SIM Image Showimg 5 Phase Translations Per Grid Rotation and 3 Rotational Angles

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

THOR - Nikon Structured Illumination Super-resolution - Raw Data of 3-D SIM Image Showimg 5 Phase Translations and 3 Rotational Angles (Close Up)

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

THOR - Nikon SIM/A1

BLACK WIDOW - Nikon Ti Eclipse spinning disk with FRAP

Lasers

405, 488, 561, 640 (nm)
Quad used for excitation transmission in photobleaching or photoactivation experiments is 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:

4x 0.13NA Plan Fluor WD 17.2 (mm)
60x 1.27NA water SR Plan Apo WD 0.17 (mm)

Andor iXon Ultra 897 EM-CCD
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 nm Coherent OBIS laser (50mW)
SOLA Light engine
Motorized X, Y, Z
Nikon Perfect Focus System
TOKAI-HIT Cell Incubator; 1 X 35 (mm) dish or chamber slide (coverslip bottom).

NIS Elements

Lamin A Laser Depletion

“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 (Left View)

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 (Left View)

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

Black Widow User Guide

BLACK WIDOW - Nikon SDC

Chronos - Yokogawa & Olympus CellVoyager™ CV1000 spinning disk scanner

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)
20x 0.45NA LUCPLFLN 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)

Hamamatsu Photonics | ImagEM X2 EM-CCD Camera C9100-14 High Resolution Format With 1024 x 1024 Pixels

This is a 1024 x 1024, 16 bit detector with 13 x 13 micron 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

Chronos is equipped with an ultra-sensitive, high-resolution mode Hamamatsu EMCCD camera and a Yokogawa X1 spinning disk with the option of dual microlens-enhanced Nipkow scanning disks with pinholes of 50 μm or 25 μm; disks can also be removed for widefield imaging. Zero Drift (Olympus) autofocusing and software focusing. This microscope has minimal phototoxicity and photobleaching, and is 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 (Stage Out Front View)

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)

CHRONOS - Yokogawa/Olympus CV1000

IRON MAN - Zeiss 510 laser scanning confocal

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

Zeiss Zen 2009

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

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

Iron Man User Guide

IRON MAN - Zeiss 510 LSM

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Confocals

THOR - Nikon A1 laser scanning, structured illumination and TIRF

Lasers:

Filters for widefield:

N-SIM certified objectives:

Standard objectives:

Available upon request:

Laser Scanning Confocal:

N-SIM:

BLACK WIDOW - Nikon Ti Eclipse spinning disk with FRAP

Lasers

Standard objectives:

Available upon request:

Chronos - Yokogawa & Olympus CellVoyager™ CV1000 spinning disk scanner

Lasers

Standard objectives:

Objectives available upon request:

The nitty gritty

Standard sample holders

Environmentally controlled chambers

Acquisition

Viewer

IRON MAN - Zeiss 510 laser scanning confocal

Lasers

Filters

Standard objectives