Core Capabilities

The EML provides Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) services as well as expertise in the following specialized areas: Immunoelectron microscopy, cryo-electron microscopy, negative stain analysis, ultrastructure analysis and virus particle characterization.

Instrumentation

The EM Lab suite includes four transmission electron microscopes and one scanning electron microscope. Supporting equipment includes an FEI Vitrobot, four ultramicrotomes, a high-resolution sputter coater and light microscopes with digital capture capability.

Established Technologies

• transmission electron microscopy (TEM)
• scanning electron microscopy (SEM)
• cryo-electron microscopy
• electron tomography
• nanoparticle analysis (soft matter and hard matter)
• elemental analysis by energy dispersive x-ray spectroscopy
• selected area electron diffraction
• consultation/training
• image data storage and access

User Guidelines

EML services are available to all CCR Investigators. To request services from this CCR-dedicated core facility, you must submit your requisition through NAS.

Established Technologies

• Anatomic Pathology
• Toxicologic Pathology
• Mouse Embryology
• Genetically Engineered Mouse Phenotyping
• Hematology and Clinical Chemistry (non-human)
• Tissue Embedding/Block Preparation (paraffin, OCT, Resin)
• Tissue Microarray Production
• Microtomy (paraffin, frozen, vibratome). Serial and step sectioning.
• Histological Stain
• Molecular Pathology: immunohistochemistry (IHC); in situ hybridization (ISH)
• Nucleic Acid Isolation from fresh/frozen/fixed tissues, paraffin blocks, cut slides, paraffin cores/ribbons
• Laser Capture Microdissection (LCM) and Laser Cutting (LC)
• Digital Image Capture (Whole slide Bright field / Fluorescent)
• Digital Image Analysis via Indica Labs HALO

User Guidelines

To request services from this core facility, submit a requisition through the NCI at Frederick Accessioning System NAS.

Established Optical Microscopy Technologies

• Bright-field, phase, differential interference contrast (Nomarsky) imaging
• FRAP
• FRET
• Multi-color, high-resolution fluorescence image acquisition of fixed and live specimens (up to 3 colors).
• Ratio imaging of calcium ions and other elements
• Time-lapse imaging of live cells
• Image database server for storing/archiving

Advanced techniques are undertaken in collaboration with OMAL scientists

• Second-harmonic generation imaging microscopy (SHIM)
• Small animal microscope imaging
• FCS/RICS
• FLIM
• TIRF
• SIM
• PALM/STORM
• Light Sheet Microscopy
• Customized image analysis

Training and Scheduling

• Please contact Kim Peifley at 301-846-6561 to schedule a training session.

Developing Technologies

• OMAL technology development projects collectively advance multi-functional microscopy for a comprehensive understanding of molecular interactions in living cells.

User Guidelines

OMAL provides a broad range of training and support to CCR investigators which include experiment design, image acquisition, and quantitative analysis. We provide basic support and training with the aim of individuals becoming independent users. This service is free-of-charge to the NCI-CCR community. Projects involving OMAL acquiring new capabilities, are high risk in nature and/or involve significant OMAL resources, require consultation with OMAL.

Established Technologies

• multi-labeling
• fixed and live
• 2 photon
• SHG
• FRET
• FRAP
• FLIM
• time lapse with heat, humidity, CO2, and multi-position
• tiling

Instrumentation

• Nikon SoRa super-resolution spinning disk microscope
• Zeiss LSM 780 for higher sensitivity confocal imaging (GaAsP detector)
• Zeiss LSM 780 ELYRA for fixed cells  super-resolution SIM imaging 
• Zeiss LSM 880/Airyscan for fixed and live cells confocal, super-resolution, & two-photon imaging
• Zeiss Lighstsheet Z.1 for multiview imaging of large  specimens

Developing Technologies

• Intravital imaging
• Photoactivation

Established Technologies

HiTIF Core provides infrastructure and expertise to NCI intramural scientists for:

• Fully automated, high-throughput fluorescence and bright-field microscopy (Yokogawa CV7000, Yokogawa CV8000)
• High-Content Analysis (HCA) to measure up to hundreds of cellular features from microscopy images
• Assistance with miniaturization of cell-based assays in 96-well and 384-well microplates
• Robotic sample preparation, reagent addition, and plate washing
• Characterization of cellular mechanistic pathways using RNAi- and CRISPR/Cas9-based screens
• High-throughput live-cell fluorescence microscopy
• Automated tracking and image processing of subcellular objects in live cells

Developing Technologies

• Third-generation automated microscopy instrument optimization/development
• Automated RNA in situ hybridization methods for single molecule detection of endogenous genomic DNA loci, or mRNA transcripts
• 3D HTI assays
• Deep Learning methods for cellular segmentation and classification of cellular objects

Usage Guidelines

CCR Investigators can request a STARS subsidy to partially defray costs associated with the development and execution of HTI projects at HiTIF.  OSTR provides 50% subsidy towards smaller, pilot projects through the HiTIF.  To request subsidies go to the OSTR Subsidy website. Subsidies for larger projects will be considered through the RRS (Resource Request System) website. When logged into the RRS select “OSTR Technology Supplement.” 

Established Technologies

Quantitative analysis of many physiological and functional parameters, including anatomical and physiological tumor volumes, biodistribution,
tumor vasculature and perfusion, tracer kinetics (pharmacokinetics), blood flow, tissue Doppler, internal radiation dosimetry, metabolic imaging, and cardiac function.

Technologies

• X-ray CT
• MRI
• MRI/PET
• MRSI: hyperpolarized 13C metabolic imaging
• nuclear: PET/CT
• nuclear: SPECT/CT
• ultrasound
• ultrasound image guided injections
• optical: bioluminescence
• optical: fluorescence

Developing Technologies

The SAIP develops new acquisition and image analysis techniques based on the requirements of an imaging project.

Past developments included

• Dynamic Contrast Enhanced-MRI
• Ultrasound techniques using non-targeted microbubbles for determining tumor perfusion
• Ultrasound image guided injections
• Virtual MRI colonoscopy
• Hyperpolarized [13C] Pyruvate for MRI spectroscopy metabolic studies

User Guidelines

Services are available to all NCI investigators located at the Frederick and Bethesda campuses. Excess capacity can be made available to other NIH Investigators on a case-by-case basis. To request services from this NCI core facility, you must submit an SAIP imaging submission: https://ncifrederick.cancer.gov/Lasp/saip/Wizard/Requests/Step1Wizard.aspx.  After the ACUC approves the imaging submission, a NAS (https://ncifrederick.cancer.gov/services/accessioning/) is submitted.

Established Technologies

• Live cell imaging
• Equipment includes a Leica SP8 laser scanning confocal microscope, a spinning disk confocal microscope and a Nikon epifluorescence/TIRF microscope. We also have 4 imaging processing stations.

Developing Technologies

• We are developing protocols for madSTORM for use on our TIRF instrument.

User Guidelines

Facility staff provide training on all the instruments for members of LCMB. We also provide training for outside users who need to use the spinning disk confocal microscope or the Nikon TIRF instrument. We do not provide any training for any Zeiss laser scanning confocal microscope, although we do allow experienced outside users access to
our instruments. Only LCMB members can use the instruments at times when the Facility staff are not present (i.e., evenings and weekends).

Established Technologies

• Cardiovascular Phenotyping
• Metabolic Phenotyping
• Pulmonary Phenotyping
• Neuromuscular Phenotyping
• Behavioral Phenotyping
• Exercise Physiology
• Echocardiography
• In vivo functional vascular phenotyping
• Advanced Imaging Modalities (whole body high frequency ultrasound imaging, vascular imaging)

User Guidelines

Investigators who are interested in working with NHLBI Murine Phenotyping Core should contact Dr. Danielle Springer at ds628k@nih.gov

Established Technologies

Zeiss AxioObserver.Z1 epifluorescent microscope

• 2D and 3D imaging of live and fixed specimens
• Phase contrast, DIC, and multi-fluorescence imaging
• Equipped with Hamamatsu OrcaFlash4.0 camera and Zen Blue software

Zeiss Axio Scan.Z1 slide scanning microscope

• Fully automated slide scanner able to create brightfield and fluorescent virtual slides
• Equipped with a Colibri 7 LED source with a wide range of dye combinations available
  
• Objectives available include 2.5x, 5x, 10x, 20x, and 40x
• Equipped with Hamamatsu OrcaFlash 4.0 camera
  
• Images are acquired and analyzed using Carl Zeiss Zen 2.3 software

Zeiss LSM 780 confocal microscope

• Equipped with a stage top environmental chamber for live cell imaging
• 7 laser lines (405nm, 458nm, 488nm, 514nm, 543/561nm, 594nm & 633nm)
• Enables 3D reconstruction and animation of photo sections generated from thick specimensvvvv
• Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Loss
  in Photobleach (FLIP) to study dynamics of molecular mobility
• Examine molecular interactions using Fluorescence Resonance Energy Transfer (FRET)

Nikon Eclipse Ti2 widefield microscope

• Optimized for single molecule RNAFISH imaging
• Equipped with a Photometrics Prime BSI sCMOS camera and Lumencore SOLA SE 365 FISH light engine
  
• Motorized scanning stage with Z piezo insert for fast Z and multi position time lapse imaging
  
• Objectives available include a 10x, 20x, 40x Oil, 60x Oil, 100x Oil
  
• Filter cubes are narrow band optimized for RNAFISH imaging using DAPI, FITC, Gold, Red, and Cy5
  

• Images are acquired and analyzed using Nikon Elements software
  

  
  

• Carl Zeiss LSM780 confocal microscope
• Carl Zeiss AxioObserver.Z1 epifluorescent microscope
• Carl Zeiss AxioScan.Z1 slide scanning microscope
• Nikon Eclipse Ti2 widefield microscope
  

User Guidelines

The Core is open to all NCI intramural researchers, although LGCP scientists have priority use of the equipment in the facility. Researchers from other institutes are also welcome to use the facility. Facility staff provides individual training for all new users. Unless special arrangements are made, the equipment in the facility may be used Monday to Friday, 10:00 am – 6:00 pm

Established Technologies

Facility personnel are available for consultation about the design of imaging experiments and/or the analysis of image data. Special expertise is available for assistance with live cell imaging as well as deconvolution microscopy. Facility personnel are also experienced in the other techniques listed below and can provide expertise and software for quantitative analysis of images.

• 2D or 3D imaging of fixed and living specimens
• High-resolution imaging using confocal microscopy or deconvolution
• Protein colocalization
• Fluorescence photobleaching techniques, such as FRAP
• Analysis of protein interactions by FRET
• Super Resolution (SIM; STORM)

Developing Technologies

• Quantification procedures for FRAP, FCS (TICS) and procedures for cell mobility tracking
• Single molecule tracking inside of cell nuclei is under development.

Instrumentation

• Delta Vision Elite wide-field deconvolution microscope
• Delta Vision Core deconvolution microscope
• ELYRA Super Resolution Microscope
• Carl Zeiss LSM780 Microscope

User Guidelines

Open to all NCI intramural researchers. Researchers from other NIH institutes are also welcome to contact us.

Both new and experienced users have used the Facility to carry out short- and long-term microscopy research projects.

New Users interested in using the core’s microscopes must first contact Dr. Karpova.
Following the initial request, Dr. Karpova will meet with the user and his/her principal investigator to discuss the research project. Advice will be provided to the user regarding the advantages/limitations of the different forms of microscopy available, both in this core and elsewhere. The staff will also help the user develop an overall research strategy, design experiments, and solve specific experimental problems.

Consultation is also available regarding image analysis and interpretation and/or selection and purchase of microscopy hardware and image processing software.