The Hematopathology Section of the Laboratory of Pathology, NCI, offers specialized testing to assist in the diagnosis of lymphoproliferative disorders and immunodeficiency states. Testing is done in collaboration with the Immunohistochemistry Service of the Specialized Diagnostic Unit, and the Flow Cytometry Service. Specimen Collection and Handling

Specimen Collection and Handling

NIH Biopsy Specimens

1. Tissue biopsy specimens should be submitted to the Hematopathology Laboratory, Building 10, Room 2N110, within two hours of surgical removal.

2. Specimens should be submitted unfixed, in a sterile container, suspended in sterile normal buffered saline or tissue culture media (RPMI). The container should be labeled with the patient’s name, NIH identification number, and the identity of the tissue biopsy site. The specimen should be adequate in size to permit preparation of routinely fixed and processed material for histological examination, as well as frozen section histochemistry and/or flow cytometry.

3. A CRIS Requisition for Anatomic Pathology Services must be completed for all submitted specimens, according to NIH guidelines. Under "Special Instructions", indicate the need for Lymphocyte Surface Marker studies and state the NIH protocol and need for evaluation of particular antigens, if required; e.g. please evaluate for expression of CD20 or CD25.

4. In order to facilitate expeditious processing of fresh tissue specimens, biopsies may be delivered in person to the Hematopathology Section Laboratory by NIH clinical staff or the NIH Tissue Procurement Nurse prior to completion of the CRIS requisition. In such instances, the NIH protocol and need for special studies may be communicated verbally. The CRIS requisition should be completed as soon as the operative procedure is completed, within 1 hour.

5. Physicians are encouraged to consult with the staff of the Hematopathology Section regarding any specialized testing required. (Dr. Elaine S. Jaffe, 496-0183, or Dr. Stefania Pittaluga, 402-0297). Questions regarding delivery of specimens may also be directed to Medical Technologist, Theresa Davies-Hill, 496-1567. The Hematopathology Fellow on Service may be reached through the Section Office, 496-0183. The Hematopathology Fellow will assist in the preparation of specimens for analysis. Therefore, special requests should be discussed with the Hematopathology Fellow prior to submission of the tissue specimen.

6. Results of Immunohistochemistry will be considered in the context of the routine histological findings, and included in surgical pathology report.

Processing of Outside Fresh Lymph Node Biopsy
Specimens for Immunohistochemistry for Protocol Evaluation

Under certain circumstances, biopsies may be performed outside of the NIH and submitted for surface marker analysis for NIH protocol evaluation. Fresh or snap frozen tissue specimens should be accompanied by routinely processed slides and/or paraffin blocks for routine histological evaluation, in concert with special studies.

If the biopsy is performed in the Washington, D.C. area, it may be submitted in sterile saline or tissue culture media, according to the guidelines outlined above for NIH biopsy specimens. The tissue should be delivered to the Hematopathology Section, Building 10, Room 2B42, within 6 hours of removal. If it cannot be submitted fresh within the time limit, it may be snap frozen according to the procedure listed below.

1. Intact lymph node (or other involved site) should be sectioned at 2-3 mm intervals.

For immunohistochemical confirmation of phenotype in a patient with an established diagnosis, needle core biopsies may be used if an excisional biopsy cannot be performed. LP/NCI must have the original diagnostic biopsies for review, if needle biopsies are to be used for protocol evaluation.

2. A representative cross section of the lymph node should be snap frozen in OCT embedding compound (using a cryomold, if possible).

3. Freezing should be performed in liquid nitrogen bath or freezing bath of dry ice and 2-methyl butane (temperature -70 °C or less).

4. Wrap frozen block in aluminum foil and place inside zip lock bag. Label specimen with patient's name and institutional surgical pathology number. Hold at -70° C until shipping.

5. Ship specimen by overnight express on dry ice. Pack securely in styrofoam shipping container containing 15-20 lbs of dry ice.

6. In addition to snap frozen specimen, submit:

1. H&E stained slides of fixed, paraffin embedded tissue to be retained by NCI.

2. Representative paraffin block or 12 paraffin embedded recut slides on charged slides for immunohistochemistry/ in situ hybridization.

3. A copy of the outside surgical pathology report for all specimens submitted.

In order to avoid delay in performing immunohistochemical studies, please submit the above materials at the same time as the frozen tissue. We cannot perform immunohistochemical studies until we have reviewed the routine H&E stained slides of the submitted material.

7. Notify NIH staff physician in advance of shipping, and indicate that someone will be available to receive tissues. .

8. Complete an NIH Tissue Examination Form and submit with all materials to the Hematopathology Section Office, 10/ 2B42.

Drafted : November 8, 1996

Revised Procedure: April 6, 2007

Hematopathology Consultations

The staff of the Hematopathology Section provide consultation in selected cases of diagnostic difficulty. The Section is not a routine reference laboratory, and does not accept cases for routine immunophenotyping. The following is a list of instructions regarding cases to be submitted in consultation. We ask that each case be accompanied by a cover letter from a physician involved in the patient’s care (Pathologist or Clinician), and a copy or copies of the outside pathology report(s):

The cover letter should provide:

_____ 1. Pertinent clinical information (Brief clinical history)

_____ 2. The reason for the consultation

_____ 3. Specific questions to be answered

_____ 4. The referring pathologists working diagnosis or differential diagnosis

In addition, please submit representative H&E stained slides along with the paraffin block. For PCR studies to detect clonal Ig or T-cell receptor gene rearrangements, we require a formalin (not B5) fixed block. If a paraffin block cannot be submitted, please submit at least 12 unstained slides on charged slides.

 The Hematopathology Section will retain representative H&E stained slides on all cases submitted in consultation. It is preferable to submit recut slides, rather than originals, as we cannot take responsibility to return the original slides after our review. Outside special stains and immunohistochemical slides will be returned to the contributing laboratory. In selected cases if recuts cannot be prepared due to limitations in the biopsy material, original slides will be returned.

 Please direct the materials to

Dr. Elaine S. Jaffe
Building 10, Room 2B42
10 Center Drive , NIH, MSC-1500
Bethesda , MD 20892-1500
Office Phone: 301-496-0183
Office FAX 301-402-2415
Email