October 23, 2014

CMF Guide

The Central Microscopy Facility supply year round and summer investigators as well as courses with instrumentation, some owned by MBL and some on loan, so please read through this guide to familiarize yourself with the Facility. Feel free to contact the Facility personnel for more information. Some of the CMF rooms are on the MBL One Card access system. You will need to get CMF authorization for your card to work on those CMF doors.

Scheduling Time and Trainings:
We use an on-line system by Stratocore for scheduling time and for setting up training sessions on individual microscopy systems. The address is: www.ppms.info/mbl

TEM JEOL JEM-200CX – 80-200KV TEM equipped with:

  • -Low magnification mode (+/- 60 degrees tilt and 300 degree rotation)
  • -standard specimen cartridge and goniometer specimen cartridge
  • -AMT digital side mount and bottom mount cameraThe charge is $30 per hour.

SEM Zeiss Supra40VP – 0.1 – 30kV SEM equipped with:

  • -field emission source for high resolution
  • -digital image acquisition
  • -Oxford Instruments Inca Synergy 350 Energy Dispersive X-ray (EDS) microanalysis system
  • -HKL Premium electron backscatter diffraction (EBSD) system
  • -Leica cryo preparation and cryo stage components.The charge is $30 per hour for standard SEM and an additional $20 per hour for cryoSEM and EDS.

LIGHT OPTICS LABORATORIES - including widefield, confocal and image analysis:

Microscopic and photomicrographic equipment is available year round some of it through the courtesy of Carl Zeiss, Inc. The latest in optical microscopy equipment for such techniques as phase, dark-field, fluorescence, polarization, reflected light, and DIC are available.

Widefield Zeiss AxioImager.Z2
Upright microscope set up for transmitted light techniques such as brightfield, phase and DIC. Also set up for epi-fluorescence with a Colibri LED illuminator and mercury halide lamp. Includes a motorized stage, HRc and HRm digital cameras as well as the CRI Nuance FX system. Managed with Zeiss Zen software for acquisition and analysis.

CRI Nuance FX
The Nuance is spectral imaging system installed on the AxioImager Z2 microscope. Provides fluorescence and transmitted images consisting of a spectral data cube. Possible to separate closely overlapping signals with linear unmixing and auto fluorescence subtraction.

Zeiss ApoTome
The ApoTome is an innovative structured illumination slider that can be installed on the AxioImager Z2 microscope. Provides improved image quality and enhanced contrast in conventional fluorescence microscopy. Deblurred 2D planes, even with thick specimens. Utilizes a high-end technique for artifact-free structured illumination. Flexible technique used for the entire spectrum of potential fluorochromes with the appropriate filter cubes.

LC-PolScope
The CRI LC-PolScope unit can be mounted on the Imager. It is a quantitative imaging system that measures and displays the birefringent structure in transparent objects with computer controlled polarized LM techniques.

Widefield Zeiss AxioImager.M2 Upright microscope set up for brightfield and DIC transmitted light. Also set up for epi-fluorescence with a mercury halide lamp. The system has the MRc color and MRm monochrome cameras and AxioVision software.

Widefield Zeiss Axiovert 35
Inverted microscope set up for brightfield transmitted light and standard epi-fluorescence. MRm camera and AxioVision software allow image capture.

Stereo Zeiss Discovery V20
Stereo dissecting microscope set up for transmitted light techniques with a VisiLED base and reflected light with a fiber optic illuminator. Includes a motorized stage, fluorescence module, MRc camera and controlled with AxioVision software.

Standard light microscopy and image analysis is charged at $10.00 per hour.

MICRODISSECTiON and OPTICAL LASER TWEEZERS MICROSCOPY SYSTEM:

Zeiss Combi PALM Microdissection and optical laser tweezers paired with the Axiovert 200M inverted microscope. Equipped with a digital camera. This unit is capable of catapulting (dissecting) single cells off a cover slip and into microtube caps. It is also capable of manipulating cells through the medium with a laser beam.

The charge is $20 per hour.

CONFOCAL MICROSCOPY SYSTEMS:

Zeiss LSM-710 NLO– Inverted
The MBL has a Zeiss LSM-710 NLO laser scanning confocal system. The Zeiss 710 LSM has three independent simultaneous spectral detection channels for reflection and fluorescence and an extra detector for transmitted light. System is capable of confocal imaging using 7 visible lines – 405, 458, 488, 514, 561, 594 and 633 nm as well as two photon excitation using the Coherent Chameleon Vision II laser. The non-descanned system includes a 2 channel GaAsP detector for reflected light and a 2-channel module on the transmitted path. This system operates with the ZEN software interface. The Zeiss 710 LSM is configured on an inverted Observer Z1 microscope with a motorized stage along with a variety of objectives and a complete incubator chamber system.

Zeiss LSM-510 NLO META – upright
The MBL has a LSM-510 NLO with META detector laser point scanning confocal system configured on a fixed stage Axioskop 2FS microscope with a variety of objectives and a custom motorized stage. It is configured for electrophysiology with Sutter MP-285 manipulators, Axon Instruments Digidata 1440A, Multiclamp 700A and Axoclamp-2B. The Zeiss 510 LSM has three independent simultaneous confocal channels for reflection and fluorescence and an extra detector for transmitted light. System is capable of confocal imaging using 6 visible lines – 458, 477, 488, 514, 543 and 633 nm as well as two photon excitation using the Coherent Chameleon laser. This system operates with the ZEN software interface.

Zeiss LSM-780 Quasar– Inverted
The MBL has a Zeiss LSM-810 laser scanning confocal system. The Zeiss 780 LSM has the 34 spectral detection GsAsP channels Quasar detector for reflection and fluorescence and an external detector for transmitted light. System is capable of confocal imaging using 7 visible laser lines – 405, 458, 488, 514, 561, 594 and 633 nm. Included is a module for polarization anisotropy. This system operates with the ZEN software interface. The Zeiss 780 LSM is configured on an inverted Observer Z1 microscope with a motorized stage along with a variety of objectives.

Zeiss Cell Observer SD – Inverted
The MBL has a Zeiss Cell Observer SD spinning disk confocal system. The SD has a Photometrics Evolve 512×512 camera. System is capable of confocal imaging using 4 visible lines – 405, 488, 561 and 635 nm This system operates with the Zeiss Zen software interface. The Zeiss SD is configured on an inverted Observer Z1 microscope with a motorized stage along with a variety of objectives, an MRm camera and a complete large incubator chamber system. It is mounted with electrophysiology equipment and a FRAP module.

REPRESENTATIVE FOR SEASONAL LOAN:

Zeiss LSM 700
Zeiss has provided the LSM 700 laser point scanning confocal system capable of triple label fluorescence and high resolution DIC. The 700 has two independent confocal channels for reflection and fluorescence and an extra channel for transmitted light. Laser lines are 405, 458, 488, 514 and 543 nm. The Zeiss LSM 700 is configured on an inverted Observer microscope with a variety of objectives available. The system has image processing, colocalization analysis, quantitative measuring software and 3D rendering.

Nikon A1R
Nikon has provided the A1R resonant scanner laser point scanning confocal system capable of triple label fluorescence and high resolution DIC. The A1R has four independent confocal channels for reflection and fluorescence and an extra channel for transmitted light. Laser lines are 405, 457, 488, 514 and 561 nm.

The confocal microscope use is charged at $28.00 per hour, consumables are extra.

SUPER RESOLUTION SYSTEMS

Super resolution systems are available through Zeiss and Nikon. Contact Louie Kerr, Jim McIlvain of Zeiss or Lynne Chang of Nikon.

ELECTRONIC IMAGING FACILITIES:

Molecular Devices MetaMorph and Image J Fiji: Processing and analysis systems are available.

Copy stand with digital camera

Stand alone image processing stations – PC software: Zeiss LSM confocal, MetaMorph, Zeiss Vision and Zen, HKL EBSD, Fiji

Image analysis and computer use is charged at $10.00 per hour.

PREPARATIVE MICROSCOPY TECHNIQUE SERVICE

Please discuss with facility staff for support including fixation, embedding, sectioning, staining and scope time. The cost for this service, excluding scope time, is $28 per hour. Inquiries should be made regarding special technique requirements.

HISTOLOGY SERVICES – We are in the process of setting up a histology processing line. Please contact the CMF staff to discuss possible use for the summer of 2013.

PREPARATIVE AND ANCILLARY EQUIPMENT – Denton Vacuum evaporator equipped to:

  • - glow discharge
  • - stationary and tilting omni rotary evaporative coating and shadowing using carbon and metals

Leica MED 020 coating station:

  • - carbon thread and platinum sputter coating
  • - glow discharge
  • - cryo SEM prep chamber

Oxygen plasma etcher

Chemical fume hood, refrigerator, pH meter, osmometer, prep areas, Stereo viewer capable of parallax heightening, curing oven

Tousimis Samdri-780A critical point dryer with various specimen carriers (Liquid CO2 provided)

Reichert knife breaker and glass breaking pliers, dissecting microscope for block trimming, knife inspection, and specimen viewing, compound microscope, hot plates, and stains for viewing thick sections

The microtomes are located in a separate sectioning room and include:

  • -Ultramicrotomes: Reichert Ultracut E, and Leica EM Trim, (seasonal loan Leica UT7)
  • -Leica Vt1000S vibratome, Spencer sliding microtome, Sorvall JB-4 microtome and an AO 820 paraffin microtome
  • -Microm HM505N and Leica CM1850 Cryostats.There is a $10 per hour charge for microtome use.

The CMF maintains a supply of Liquid Nitrogen for purchase by the liter.

Contact Louis Kerr at Marine Biological Laboratory, Woods Hole, Ma, 02543; Phone: (508) 289-7273, Email: lkerr@mbl.edu for more information.