Services

Service Name Core Contact Description
Roller Bottle Cultures Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Core has the capability of producing large scale adherent cells cultured in either 850 cm” or 1750 cm” roller bottles grown on a roller apparatus in a 37°C warm room. This is a very suitable method when large amounts of cells are required. This method also works well with suspension cells.
Spinner Cultures Cells Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Useful for the production of large volumes of suspension cells. Vessels come in various sizes that are used in conjunction with a magnetic stirrer spinner base. The core can provide volumes from 100mls-10L.
EBV Transformations/Separation Of Whole Blood Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Cell Core can provide immortalization of lymphocytes with the Epstein Barr Virus (EBV). The core will separate the whole blood, collect the lymphocytes, infect with EBV, and establish a cell line.
Insect Cell Culture Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Core is equipped with a 27°C incubator that is an optimum temperature to support the growth of insect cells.
Mycoplasma Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The core routinely performs a direct and an indirect method of testing for Mycoplasma. The following 2 tests are done in parallel.
Eradication Of Mycoplasma From Cell Cultures Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The core will test for and eradicate mycoplasma from your cells. If mycoplasma contamination is present in your cultures the core will begin to eradicate using specialized antibiotics. This will take several weeks to complete. At the end of the treatment the core will provide clean cells growing in culture as well as several frozen vials.
Cryogenic Storage Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Core can accommodate the storage of cryovials. The core offers this service to LRI researchers for backup storage of precious cell lines.
Serum Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The core screens FBS from different sources for optimum cell growth. The batch that proves most compatible to a broad range of cell types is provided for LRI researchers.
Cell Culture Training Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Core provides training on good lab practice to new researchers. The training can be tailored to individual needs and includes aseptic technique and culturing and maintaining cell lines.
Sterility Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Core offers sterility services using in-house prepared broths along with regular mycoplasma and endotoxin testing. Quarterly testing results are available.
Fusion Of Cells Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

Fusion of spleen cells from chosen mouse to myeloma cell line. Plating of fused cells and collection of media samples for screening by the investigator. Includes labor and supplies. This phase goes 4-6 weeks.
Cloning Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

Limiting dilution cloning of five chosen hybridoma cell lines. Collection of media samples for screening by the investigator. Minimal scale up and preparation of frozen stocks (5 vials) of clones are completed by HCF. Includes labor and supplies. This phase goes 4-8 weeks. Freezing Cells Cells from culture expansion associated with mAb production can be frozen down for storage in Liq N2. The freezing media is 90% FBS:10% DMSO and the cells are at a concentration of 2-4 x 106 cells/ml.
Mouse Injections Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

We will perform intraperitoneal injections of antigen into mice to elicit an immune response for the production of monoclonal antiodies.
Purifications Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

We can purify monoclonal or polyclonal antibodies using Protein G or an epitope specific affinity column
ELISA Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

We can perform antigen-capture or sandwich type ELISA’s to measure antibody levels in tissue culture supernatant.
Western Blot Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

We will run the SDS PAGE gel, transfer and detect using investigator provided primary antibody plus ECL reagent.
Antigen Conjugations Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

Peptide conjugation to KLH by a glutaraldehyde protocol
Large-scale Antibody Production Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

The Hybridoma Core uses a static cell culture system – the Integra Flask – to produce high concentration (>.5mg/ml) monoclonal antibodies. This can be done in serum-free or using ultra-low IgG/IgM serum. Yields can be as high as 100mg/month/flask.
Large-scale Antibody Purification Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

We can purify up to 80mg of IgG from one sample using a protein G column
Freezing Cells Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

The Core will freeze back hybridomas in freezing media from actively growing cells.
Liquid Nitrogen Storage Of Cells Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

Storage for cloned cell lines. The stored cell lines must be mycoplasma free.
Post-translational Modification Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Identification of modification sites from either an in-gel or in-solution protein digestion
2D-Gel-Sypro Ruby Stain Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

2D gel of a single sample with fluorescent staining
Molecular Weight Analysis Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Determination of the molecular weight of a peptide or protein.
Protein Identification Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Identification of proteins from either a 1D or 2D gel band or from proteins in solution
Cell Culture Media Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

A growth medium to support the growth of cells (i.e. RPMI and DMEM)
Insect Media Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Media for insect cell culture
Specialty And Custom Media Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

A custom recipe prepared according to researchers instructions or guidelines
Bacteriological Media Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Media used for the growth of bacteria
LB Broth Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Luria Broth, a nutritionally rich medium used for the growth of bacteria
Buffers Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

A buffer is an aqueous solution that has a highly stable pH. (i.e. Phosphate and Tris Buffered Saline)
LB Agar Plates Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Luria Broth agar plates are typically used as a growth substrate for the culture of bacteria. Selective growth compounds may also be added to the media, such as antibiotics. (i.e. Ampicillin and Kanamycin) Custom plates are also available.
Sterility Testing Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Verifying the sterility of our products or yours through QC broths, endotoxin and Mycoplasma testing
Endotoxin Testing Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The Endoscan V software system uses Kinetic Turbidimetrics to provide quantitative Endotoxin results for in-process and end product samples. The assay sensitivity available for use is 0.06EU/mL using a standard curve of 5-0.05 EU/mL. The Second method uses an Endosafe-PTS- A rapid, point-of-use test system that utilizes Limulus Amebocyte Lysate (LAL) reagents in a test cartridge with a handheld spectrophotometer. The PTS can effectively be used to obtain fast; quantitative LAL test results in about 15minutes.
FBS- Heat Inactivated Or Regular Media Preparation Core

Media Preparation Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Fetal Bovine Serum, the most widely used serum-supplement due to it’s very low levels of antibodies and the fact that it contains more growth factors, allowing for versatility in many different cell culture applications.
Nucleic Acid Quality/quantity Assessment Agilent Bioanalyzer Chip, Nanodrop And Qubit Services Genomics Characterization of the integrity of RNA and DNA samples using Agilent Bioanalyzer chips. Samples may be destined for whole genome gene expression, genotyping, next gen sequencing library preparation and sequencing. Sample quantification via Nanodrop and Qubit are also offered.
Clinical Research Coach Translational Research

Translational Research

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

Help with IRB questions including webkit
Experienced in not only writing consents but in contacts for contacting subjects
How to approach your research subjects
Capillary Sequencing Genomics The Genomics Core is equipped with an Applied Biosystems 3730xl DNA Genetic Analyzer and employs experienced, well-trained personnel to handle sequencing projects. Information on how to prepare and submit samples, how to access and interpret the results is given in more detailed web pages.
High Throughput Sequencing Ilumina Genomics
Nucleic Acid Shearing Covaris Services Genomics Nucleic acid fragmentation is a crucial first step in NGS sequencing workflow. Covaris S220 shears DNA without GC bias or thermal damage. The Adaptive Focused Acoustics™ (AFA) technology is firmly established as the fragmentation method of choice for NGS library generation.
3-D Microscope Imaging Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

"Optical sections" or "Z Stacks" of samples can be obtained on confocal microscopes. The stacks can be reconstructed with software that allows 3-D visualization and analysis.
Confocal Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Laser-based confocal microscopes allow us to focus on a thin "optical section" within a sample, thus removing the out-of-focus light that comes from other layers of the sample. This offers not just a clearer image, but clarifies the signal penetration beyoud the cell surface. Both samples on slides and live samples can be examined.
Cryosectioning Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Blocks of frozen tissue are cut onto slides with a cryotome.
EDAX Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Determination of the elemental composition of an electron microscope sample
Electron Microscope (Scanning And Transmission) Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Use of electron beam (rather than photons of light) to scan the surface of or pass through a sample to achieve ultra-high magnification
Elemental Analysis (with SEM) Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Determination of the elemental composition of an electron microscope sample
EM Sample Preparation Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

The preparation of a sample for cutting and staining that will allow for it to be observed in a transmission electron microscope
Fluorescence Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Microscopes with powerful lamps and special color filter cubes allow the imaging of fluorescently tagged speciments — both on slides and in wells, dishes and flasks.
Frozen Sectioning Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Blocks of frozen tissue are cut onto slides with a cryotome.
Histology Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

The processing, embedding, cutting and staining of tissue for observation in a microscope
Image Analysis/Quantitation Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Various software programs allow microscope images to be examined for data such as area, intensity, volume, velocity, trajectory, etc. as required for 2-D, 3-D, and time-lapse experiments.
Immuno EM / Immunogold Labeling Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Pre- and Post-Embedding Immunogold Labeling for transmission electron microscopy
Immunohistochemistry Immunohistochemistry

Immunohistochemistry

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

This chemical technique allows us to visualize the expression levels and distribution of specific proteins within cells and tissues.
Infrared Scanner (Odyssey) Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Infrared Scanning of gels, membranes, slides on Li-Cor Odyssey
Laser Capture Microdissection Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Use of microscope and a laser to cut and collect individual cells or small sections of tissues or cultures.
Light Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Samples can be viewed on a microscope by allowing light to pass through a sample (transmitted light) or to shine on it (incident light, as in fluorescence imaging).
Live Cell Imaging Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Inverted microscopes allow the imaging of live cells in culture acquiring either still photos or time-lapse movies.
Multi-Photon Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

A multi-photon microscope allows deeper penetration of light into a sample (up to 500um rather than the 100um of standard confocals) and can be used for tissue slices or anesthetized animals.
Plastic Embedding Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Special form of histological tissue preparation that uses plastic resin rather than paraffin
Spinning Disk Confocal Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

This technology for imaging live samples cuts down on photodamage in time-lapse experiments and allows three-dimensional imaging.
Stereomicroscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

An automated dissecting microscope with color digital camera allows the imaging of large unmounted samples with brightfield and/or fluorescence illumination.
Thick Sectioning (for EM) Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Thick sections in the 1-2 µm range can be stained and viewed in a light microscope to determine the right area of the specimen for ultra-thin sectioning.
Thin Sectioning (for EM) Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

The embedded sample must be cut with a diamond knife into extremely thin slices for viewing in the electron microscope. Ultra-thin sections range from 50-100 nm in thickness.
Tissue Processing Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

The preparation of tissue for cutting and staining that involves dehydration and infiltration with paraffin or plastic
Tissue Staining Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Use of various dyes to render tissue visible and to mark particular features
Time-lapse Imaging Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Inverted microscopes allow the imaging of live cells in culture over a determined period of time and at set intervals, producing time-lapse movies.
Whole Slide Scanner (microscope) Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

A large region of interest - or even the whole surface of a slide - can be imaged on a special scanner.
Preclinical Research Facilities Translational Research

Translational Research

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

AFIC/ GCIC Operating Theater Two Theaters are Available
Rodent Survival Surgery Procedure Room Translational Research

Translational Research

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

Two bays with surgical microscopes and machines to administer volatile anesthetics (isoflurane).Vivid 7 echocardiography machine.
Phlebotomy Services Translational Research

Translational Research

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

Phlebotomy services are available free of charge. Blood draws of up to 100 cc can be performed by trained personnel. We will draw according to your protocol and consents. Sample Processing and Storage We can assist with your sample processing and short term storage needs. Spinning and aliquoting of serum, plasma, whole blood, buffy coat, urine and other sample types.
Antibody Titration Immunohistochemistry

Immunohistochemistry

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Experimental determination of appropriate antibody concentration for optimal imaging
Screening Chemicals Molecular Screening

Molecular Screening

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

Stage I: establishment of the readout assay
Stage II: primary screening
Stage IV: optimizing hits, SAR
Readout Systems - Using The Synergy4 Multi-detection Multi-plate Reader Molecular Screening

Molecular Screening

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

General requirements
Optimization
Screening
Data processing
shRNA bank
Preparing Your Samples For Mycroplasma Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The cells should be grown without antibiotics for 3-4 days. Collect cells for testing by scraping the adherent cells and collecting about 5mls of cells and media. For suspension cells, grow without antibiotics and supply 5mls for testing.
Direct Mycoplasma Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

This method uses an enriched agar to support the growth of colonies. Cells and supernatant are swabbed onto agar and incubated in a modular incubator chamber. Samples are viewed microscopically every other day for 2 weeks. Mycoplasma contamination is detected by the appearance of a “fried egg “-like growth.
Indirect Mycoplasma Testing Cell Culture

Cell Culture

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

The core also uses a quick method to detect mycoplasma. This kit detects the four most common types of mycoplasma to contaminate cells. This is useful in determining the type of mycoplasma present.
2D-Gel-DIGE Analysis Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

2D gel of multiple samples in a single gel using different fluorescent stains
Isotope Enrichment Analysis Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Determination of protein turnover rates by analyzing the degree of heavy isotope enrichment
Quantitative Analysis Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Quantitation at either the protein or modification level. Can be done using either label free methods or methods that include the incorporation of stable isotopes
FACS Applications Flow Cytometry Immunophenotyping
Intracellular staining
Molecular and cellular probe analysis
Apoptosis analysis and viability detection
Detection of soluble proteins
Sorting
Automated cell counts
Consultation Flow Cytometry All investigators are encouraged to meet with the Flow Cytometry Core technologists to discuss the experiment.
Training Flow Cytometry The LRI Flow Cytometry Core offers training programs for Beginning and Advanced Flow Cytometer Operators.
Laboratory Testing Laboratory Diagnostic Core

Laboratory Diagnostic Core

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

Automated clinical chemistry assays
Drugs of Abuse/Toxicology
Specific Proteins
Metabolic
Special Chemistry
Fertility/Pregnancy
Therapeutic Drug Monitoring
ELISA based testing
Peptide Synthesis Molecular Biotechnology

Molecular Biotechnology

Smarajit Bandyopadhyay Ph.D.
Project Staff

Location:NB2-37
Phone:(216) 445-7095
bandyos1@ccf.org
Fax:(216) 444-9404

Peptides are complex molecules and each peptide sequence is unique with regard to its chemical and physical properties. Peptides are synthesized by the solid-phase method using Fmoc chemistry using a liberty peptide synthesizer(CEM Inc.) based on microwave technology and an Omega 396 multiple peptide synthesizer (Advanced ChemTech).
Targeted Metabolomics Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Small molecules (100-800 Daltons) have a variety of biological functions, serving as cell signaling molecules, as tools in molecular biology, and as drugs in medicine. Liquid chromatography on-line tandem mass spectrometry (LC/MS/MS) is the method of choice for small molecule quantitation because it provides accurate, reliable and consistent data and requires fewer specimens. We provide services using LC/MS/MS for quantitation of small molecules in complex biological matrices, such as plasma, urine and cell extracts.
Novel Compound Identification Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

Anyone isolating compounds from biological sources has an interest, first, in establishing the activity of compound, then its molecular weight, and finally, the compound’s definitive structure. We provide services using HPLC for purification of the compound interested and LC/MS/MS for identifying the molecular weight of the compound and for producing structure confirmation if needed.
Method Development Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

It is essential to provide accurate, reliable, and consistent data in analytical services. Based on the need of investigators, we provide services for developing analytical methods using a) HPLC-UV, b) HPLC-Fluorescence, and c) LC/MS/MS for analysis of endogenous compounds and xenobiotics in biological matrices like plasma, urine and tissues.
Surface Plasmon Resonance (SPR) Molecular Biotechnology

Molecular Biotechnology

Smarajit Bandyopadhyay Ph.D.
Project Staff

Location:NB2-37
Phone:(216) 445-7095
bandyos1@ccf.org
Fax:(216) 444-9404

Surface Plasmon Resonance (SPR) has been used to monitor macromolecular interactions in real time. The Biacore 3000 system is an instrument that uses SPR technology for measuring the interactions of macromolecules with each other, and with small ligands. One of the ligands is immobilized on carboxymethylated dextran over a gold surface, while the second partner (analyte) is captured as it flows over the immobilized ligand surface. Most ligands can be directly immobilized onto the surface of the chip via amino groups, carbohydrate moieties, or sulfhydryl groups. Others are immobilized indirectly through the use of biotinylation of the ligand (such as biotinylated peptides or oligonucleotides), or through immobilized monoclonal antibodies (such as anti-GST). Typical amounts of a protein ligand needed for an immobilization reaction is about 1 µg. The immobilized ligands are remarkably resilient and maintain their biological activity.
Glassware Services Glassware Core

Glassware Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Collection of glassware from labs daily
Storage of sterile glassware
Daily delivery and stocking of glassware in lab areas
Sterilization And Autoclaving Glassware Core

Glassware Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Washing and Sterilization of all types of glassware
Sterile Pipettes and Pasteurs
Autoclaving of liquids and dry materials
Washing and Sterilization of special glassware
Sterile Tips and custom tips available
Sterile DI water
Quarterly Testing Of DI Water Glassware Core

Glassware Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

DI water from throughout the Lerner complex is tested for Endotoxins on a quarterly basis. Results are available in NB1-15.
Biohazard Waste Processing Glassware Core

Glassware Core

Carmel M. Burns
Manager

Location:NB1-25
Phone:(216) 444-5814
burnsc@ccf.org

Live or contagious waste can be decontaminated by the autoclave process before disposal; this service is available through the glassware core.
Analysis Of Whole Blood CBCs Hematology Analysis

Hematology Analysis

Alan Pratt MT (ASCP)
Manager

Location:NE6-205
Phone:(216) 444-4299
pratta@ccf.org
Fax:(216) 636-2070

Absolute and percent Reticulocyte Count (Retic)
CBC plus white cell differential counts (CBC/Diff)
CBC/Diff plus retic (CBC/diff/retic)
CBC/retic
Complete Blood Count (CBC)
Integra Culture System Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

For production of mAb from cell lines This system is intended for production of 30-60mg of mAb per month in the smaller system and 100-200mg of mAb in the larger system. Ave. concentration is 1.5mg/ml. The production schedule ave. is 8 weeks.
Polyclonal Antibody Production Hybridoma

Hybridoma

Earl Poptic
Manager, Hybridoma Core and Molecular Screening Core

Location:NB1-25
Phone:(216) 445-6635
poptice@ccf.org
Fax:(216) 445-0515

This service will be offered on a limited basis depending on available housing. One rabbit per antigen. Includes animal purchase and board, 6 injections, 1 test bleed, 3 production collections, final bleed and labor for 120 days. Time course can be extended for additional fee based on animal board and labor charges. The goal is to provide a total of 100mls of serum, but this is not guaranteed. Allow 1-2 weeks for animals to arrive.
Slide Scanning Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

A large region of interest - or even the whole surface of a slide - can be imaged on a special scanner.
Two-Photon Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

A multi-photon microscope allows deeper penetration of light into a sample (up to 500um rather than the 100um of standard confocals) and can be used for tissue slices or anesthetized animals.
TIRF Microscope Digital Imaging Microscopy

Digital Imaging Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Total Internal Reflection Fluorescence with a microscope using a laser and specially designed optics
Critical Point Drying Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Process for drying a sample for scanning electron microscopy in a way that does not cause surface deformation
Glow Discharge Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Removal of positive charge on an electron microscope grid to prevent dispersion of sample
Sputter Coating Electron Microscopy

Electron Microscopy

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Samples for scanning electron microscopy are first prepared by depositing an ultra-thin layer of gold on the surface
Paraffin Embedding Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

Placement of processed sample into wax block for sectioning
Sectioning Histology

Histology

Judith A. Drazba Ph.D.
Staff, Core Director

Location:NB1-46
Phone:(216) 445-3760
drazbaj@ccf.org

The cutting of embedded tissue onto slides (also "cryosectioning")
Isothermal Titration Calorimetry Molecular Biotechnology

Molecular Biotechnology

Smarajit Bandyopadhyay Ph.D.
Project Staff

Location:NB2-37
Phone:(216) 445-7095
bandyos1@ccf.org
Fax:(216) 444-9404

Isothermal Titration Calorimetry (ITC) is the gold standard for measuring biomolecular interactions. ITC simultaneously determines all binding parameters (n, K, δH and δS) in a single experiment – information that cannot be obtained from any other method. When substances bind, heat is either generated or absorbed. ITC is a thermodynamic technique that directly measures the heat released or absorbed during a biomolecular binding event. Measurement of this heat allows accurate determination of binding constants (KB), reaction stoichiometry (n), enthalpy (δH) and entropy (δS), thereby providing a complete thermodynamic profile of the molecular interaction in a single experiment. Because ITC goes beyond binding affinities and can elucidate the mechanism of the molecular interaction, it has become the method of choice for characterizing biomolecular interactions.
CD Spectroscopy Molecular Biotechnology

Molecular Biotechnology

Smarajit Bandyopadhyay Ph.D.
Project Staff

Location:NB2-37
Phone:(216) 445-7095
bandyos1@ccf.org
Fax:(216) 444-9404

A Circular Dichroisms (CD) Spectropolarimeter (Model J-815 from Jasco) is a type of light absorption spectroscopy that can provide information on the structures of optically active biological macromolecules. CD spectra of proteins between 250 and 185 nm can be analyzed for different secondary structural types such as, alpha helix, parallel and antiparallel beta sheet, turn and other random structures.
Storm 820 Phosphorimager Molecular Biotechnology

Molecular Biotechnology

Smarajit Bandyopadhyay Ph.D.
Project Staff

Location:NB2-37
Phone:(216) 445-7095
bandyos1@ccf.org
Fax:(216) 444-9404

The Storm system is an optical scanner that produces digital images of radioactive or fluorescently labeled samples. The Storm 820 Phosphorimager is used only to scan a phosphor screen that has been exposed with a radioactive sample. Users will need to provide their own phosphor screen. One can analyze the results of a scan with ImageQuant TL which has been installed on the data acquisition computer. Storm 820 is located in NB-5, near the elevators. A sign-up log book is available next to the instrument.
Gene Expression Profiling Services Genomics
  • Clients provide the Core with arrays (human or mouse) and total RNA (minimum of 1 microgram, 10 ul @ 100 ng/ul).

The Core will perform an RNA quality control step, process the RNA samples if they pass, hybridize to the arrays and produce relevant output files (at the Cleveland Clinic the Core can install the Illumina GenomeStudio software on a PC in your lab).

Genotyping Services Genomics
  • Clients provide the Core with arrays (Infinium or GoldenGate) and DNA (generally 10 ul @ 50 ng/ul). The Core will process the DNA samples, hybridize to the arrays and produce relevant output files (at the Cleveland Clinic the Core can install the Illumina genomeStudio software on a PC in your lab).
  • Supported products: (most) Infinium and all Goldengate products. E.g.
Methylation Services Genomics
  • Clients provide the Core with arrays (Infinium HumanMethylation450) and DNA (10 ul @ 50-100 ng/ul). The Core will process the DNA samples, hybridize to the arrays and produce relevant output files (at the Cleveland Clinic the Core can install the Illumina GenomeStudio software on a PC in your lab).
  • Bisulphite conversion can also be done in the Core
  • Supported products: Infinium HumanMethylation450.
Untargeted Metabolomics Proteomics And Metabolomics

Proteomics And Metabolomics

Belinda Willard Ph.D.
Director

Location:NE1-251
Phone:(216) 444-7170
willarb@ccf.org
Fax:(216) 444-9404

The unbiased analysis of small molecules (100-800 Daltons) derived from a variety of biological matrices such as plasma, urine, and cell extracts can be performed in the Metabolomics Core. These experiments involve the extraction of the small molecule metabolites, a survey LC-MS/MS analysis, chromatographic alignment of the LC-MS data, and quantitative comparison of these metabolites across groups. The data is analyzed with two different methods. The first involves the identification and quantitation of all metabolites based on the observed m/z ratio and retention time. This analysis results in the identification of 500 to 1000 metabolites many of which are unnamed compounds. The second method of data analysis involves the comparison of the observed metabolites to an in-house metabolite library. This method of data analysis results in the determination of the relative abundance of 100's of named metabolites in these samples.