Laboratory Animal Sciences Program
LASP logo   NCI-Frederick
  
spacer
spacer
PHL PHL banner
spacer
spacer


Molecular Pathology

The Molecular Pathology Group (MPG) is a recent addition to the laboratory Animal Science Program (LASP), at the Frederick, Maryland campus of the National Cancer Institute. This Leidos Biomedical Research, Inc. group is actively working to expand high-quality, pathology-based research application support available to NCI investigators and their collaborators. Its primary role is to extend the capabilities of the existing Leidos Biomedical Research, Inc. Pathology and Histology Laboratory (PHL) core group by coordinating and performing advanced, tissue-based molecular imaging studies at the request of investigators. Our goal is simple: we want to be a flexible extension to your laboratory; your collaborating laboratory.

Click on the plus icon ( + ) to view details for each section heading.

+ or - detailsNucleic acid isolation from fixed (archival) tissues coupled with appropriate integrity analysis / quality assessment

It is often desirable to confirm intrinsic nucleic acid quality prior to committing an archive sample into a relatively expensive follow on application. The MPG can help accomplish these money and time saving tests for you by “sub-sampling” a portion of the total sample via optimized protocols and then assessing quality via an appropriate technique. For example, our in-house Agilent 2100 bioanalyzer can be used to assess quality of ng to pg amounts of total RNA via a RIN calculation assay. Similarly, in-house quantitative PCR-based technology or fluorimetry can be used to define the amount of amplifiable DNA isolated including at values below the level of detection of standard spectrophotometry.

+ or - detailsPilot study

It is sometimes desirable to perform a pilot or feasibility study on a small number of samples prior to committing to a larger study. The MPG can help accomplish this for you by assessing suitability. PCR based technology can also be adapted to produce template material suitable for whole genome amplification (WGA) prior to comparative genomic hybridization (CGH). Analogously, optimized reverse transcription can be coupled with whole transcript amplification (WTA) prior to gene expression profiling. Purified PCR products can also be utilized directly for DNA sequencing using chain termination or, in the near future, pyrosequencing approaches.

+ or - detailsNucleic acid isolation from fixed (archival) tissues coupled with appropriate integrity analysis / quality assessment and/or a pilot study (follow-on application)

It is often desirable to confirm intrinsic nucleic acid quality prior to committing an archive sample into a relatively expensive follow on application. It is also desirable to perform pilot studies (or feasibility study) on a small number of samples prior to committing to a larger study. The MPG can help accomplish these money and time saving tests for you by “sub-sampling” a portion of the total sample via optimized protocols and then assessing quality via an appropriate technique. For example, our in-house Agilent 2100 bioanalyzer can be used to assess quality of ng to pg amounts of total RNA via a RIN calculation assay. Similarly, in-house quantitative PCR-based technology or fluorimetry can be used to define the amount of amplifiable DNA isolated including at values below the level of detection of standard spectrophotometry. PCR based technology can also be adapted to produce template material suitable for whole genome amplification (WGA) prior to comparative genomic hybridization (CGH). Analogously, optimized reverse transcription can be coupled with whole transcript amplification (WTA) prior to gene expression profiling. Purified PCR products can also be utilized directly for DNA sequencing using chain termination or, in the near future, pyrosequencing approaches.

+ or - detailsAdvanced tissue processing tailored to your specific follow on application

Are you concerned that your tissue processing method may be having an unwanted influence in your gene expression profiling study? As a partner in the recently completed mouse transcriptome project responsible for eliminating this type of variation we have developed “rapid necropsy” and “tissue type specific” nucleic acid processing variations procedures optimized to retain RNA or DNA integrity. We invite you to take advantage of our experience to help guide your next project.

Pathology consultation

Do you need expert help identifying and harvesting tumor cells from within a heterogeneous micro environment? Not certain of the physical changes associated with the progression of specific cancers in different animal models? Our staff of pathologist and research technologists will work with you to identify appropriate biomarkers, define the milestone.

Novel, immunohistochemistry (IHC) assay development

+ or - detailsTissue culture cell processing

Would your group benefit from identification or creation of positive and negative control reagents suitable for IHC or ISH assay development? Interested in changes that take place to defined cell lines following introduction into an animal model or as the result of therapeutic treatment. We have significant experience in rapidly creating appropriately formatted reagents that can meet your specifications.

+ or - detailsDevelopmental dynamics

In situ hybridization assay development including: molecular probe design, generation and validation, identification / production of control tissues/cell lines, study design, ISH assay, image documentation, and pathology result interpretation.

+ or - detailsMolecular Phenotyping

Pathology driven laser capture micro-dissection (DNA and RNA)

Examples of successfully completed projects include:

  • Analysis of temporal and spatial gene expression patterns in a variety of intact tissues through immunohistochemistry, in situ hybridizations and nucleic acid analyses either alone or in combination.
  • Isolation and quantization of PCR amplifiable DNA from sarcoma biopsies
  • Quality analysis of DNA or RNA following laser capture micro dissection
  • The group also acquires, develops and has banked a range of research reagents including molecular probes, tissue, DNA, RNA, and cDNA that it can contribute to collaborative projects.

Key Technologies

    + or - detailsIntact Tissue Processing

    A complete facility for the processing of live animals and/or tissue biopsies into formatted research reagents. This includes necropsy and the production of frozen, fixed-frozen (cryoprotected), fixed “wet” tissue, and their formatting for microtomy (frozen embedded in OCT or fixed embedded in paraffin). The group also has the capacity to produce tissue micro arrays and for tissue sectioning and histological analyses including professional pathology interpretation.

    + or - detailsCell Culture

    A set of cell lines and plasmid vector constructs used for the derivation of control cell lines used in tissue based molecular imaging.

    + or - detailsNucleic Acid Isolation

    A complete facility for the isolation of nucleic acid from tissue biopsies.

Key Equipment

    + or - detailsAgilent 2100 Bioanalyzer

    A microfluidics-based platform for the analysis of DNA, RNA, proteins and cells

    + or - detailsAlpha Innotech FluorChem SP Image Doc System

    A high-performance chemiluminescence & visible Imaging System with a 12 bit, 2.0 million pixel cooled camera and ML-26 dual wavelength UV transilluminator. Includes FluorChem AlphaEase FC 32-bit software for image acquisition, enhancement, archiving, documentation, and analysis.

 

 

  National Cancer Institute (NCI)      National Institutes of Health (NIH)      Department of Health and Human Services (HHS)      FirstGov.gov  
 
     Privacy Policy  |  Developed by C&SS |  Internet Toolkit