Specimen Collection Guide

• Autopsy
• Cytogenetics
• Cytopathology
• Flow Cytometry
• Hematopathology
• Electron Microscopy
• Molecular Pathology
• Surgical Pathology

Postmortem Section - Procedures for Obtaining Autopsies or Submitting Materials for Examination

Instructions for having an autopsy performed at the NIH Clinical Center by the Postmortem Section of the Laboratory of Pathology

In general, any patient who has been seen at the NIH for any reason is eligible for an autopsy done by the Laboratory of Pathology. In addition, non NIH patients with diseases of significant clinical or basic research interest may have an autopsy done at the NIH if permission is obtained from the Chief of the Postmortem Section and the sponsorship of a principal investigator at the NIH. For an in-patient death or the death of an NIH or non-NIH patient outside the NIH:

1. Upon the death of an in-patient or notification of the death of an NIH patient outside the NIH, obtain a Death Packet from Admissions.

2. If the death falls within the jurisdiction of the Maryland State Medical Examiner, the Medical Examiner must be consulted prior to obtaining permission for autopsy. In general, patients who die from accidental causes, who arrive at the Clinical Center and are pronounced Dead On Arrival, or who die in association with a therapeutic procedure may fall under the jurisdiction of the Medical Examiner.

3. The Death Packet contains all of the necessary forms and instructions for having an autopsy performed on an NIH patient.

4. The NIH follows the State of Maryland's guidelines for consent to post-mortem examination. Permission from next-of-kin must be obtained even if the patient had a stated desire for an autopsy. The following is exerpted from the Annotated Code of Maryland, Section 5-501.

Consent for postmortem examination

(a) In general - Consent for a postmortem examination of a body by a physician is sufficient if the consent is given as provided in this section.

(b) Persons authorized to consent --

(1) The consent may be given by any one of the following persons if that person, whether alone or with another, has assumed control of the body for its final disposition:

(i) A parent;
(ii) A spouse;
(iii) A child;
(iv) A guardian;
(v) A next of kin; or
(vi) In the absence of these persons, any other person.

(2) If a person does not assume control of a body under paragraph (1) of the subsection, the consent may be given by the State Anatomy Board.

(c) Form of consent--The consent may be in the form of:

(1) A written document;
(2) A telegram; or
(3) A recorded telephonic or other recorded message

5. The patient may be transferred to the morgue from the nursing unit to await either autopsy or transfer to the responsible funeral home. The body must have proper identification (toe tag, wrist band) or the autopsy will not be performed.

6. Once permission for autopsy has been obtained, a member of the clinical care team should contact the pathology resident who will be assigned to the case, so that the clinical history and the questions to be answered may be reviewed with the pathology resident. The name and contact information of the pathology resident responsible for a new case may be obtained from the Surgical/Autopsy Pathology office (6-2441) during normal working hours. During weekday evenings and on weekends or holidays the resident on-call is responsible for performing autopsies. The name and contact information may be obtained through the NIH Page Operator.

7. The autopsy will be performed when the body, chart and permission have all arrived in the Laboratory of Pathology. Autopsies are begun routinely from 8:30 AM to 4:00 PM, Monday to Saturday and from 8:30 AM to 2:00 PM on Sundays and Holidays. Permission to perform an autopsy outside these times should be obtained from Dr. David Kleiner, Chief, Postmortem Section, Laboratory of Pathology. He can be reached by page (102-10333), by work phone (301-594-2942) or through the resident on-call.

Submission of Outside Materials for Review by the Postmortem Section

In general, the Postmortem section accepts materials for review or second opinion on patients who have died and have had an autopsy performed elsewhere.

1. The materials submitted may consist of tissue specimens, fixed or frozen, paraffin tissue blocks or stained slides. A cover letter or outside autopsy report identifying the materials should accompany the request for review. Copies of chart information, patient history summaries and laboratory data are helpful and should be made available to the Laboratory of Pathology.

2. The materials and any associated paperwork should be brought to the Surgical/Autopsy Pathology Office (Building 10, Room 2B50, 496-2441). A tissue examination request form should be filled out by the submitting physician. Frozen or Fixed tissue should be brought in sealed containers within an uncontaminated box.

3. The secretaries in the office will assist the submitting physician so that the case is appropriately handled and accessioned. The case is assigned to the resident pathologist on the postmortem service for that month.

4. If surgical or cytopathology materials are submitted with the autopsy materials, they will be separated and submitted to the appropriate section in the Laboratory of Pathology.

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Clinical Cytogenetics Laboratory Information Sheet

Laboratory Director:

Diane C. Arthur, M.D.
Phone: (301) 435-3712
FAX: ---(301) 435-3716
E-mail
Pager: 102-10334

Laboratory Staff:

Shannon I. Skarshaug, M.S. - Cytogenetic Technologist
Valerie Zgonc, CLSp(CG) - Cytogenetic Technologist
Veronica Krevinko, M.T. (ASCP) – Cytogenetic Technologist

Temporary Location: Building 10 - Room 4B50

Mailing Address:

NIH/NCI/CCR Laboratory of Pathology
Building 10 - Room 2A33
10 Center Drive
Bethesda, MD 20892-1500

Laboratory Phone: (301) 435-3711

Test Ordering: Before entering an order for testing in CRIS, we would appreciate a phone call to the laboratory to schedule the testing at least 24 hours in advance of obtaining the sample(s). Please provide the following information when ordering the test:

Patient name, ID number, and date of birth or age
Attending Physician and/or Fellow's name and phone number
Diagnosis and pertinent clinical history
Type of testing to be done
Indication for testing
Date, time, and place the specimen will be collected

We will make arrangements to pick up all specimens when they are collected.

Reporting of Results: When the analysis is complete, a final report will be typed into the SoftPath system for electronic transmission to the CRIS, and a printed report will be sent to the patient's medical record, with a xerox copy to the physician(s). If abnormal or unexpected results are obtained, Dr. Arthur will contact the responsible physician(s) by telephone or electronically.

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Flow Cytometry Procedure Manual (Specimen Collections)

1. Fluids
2. CSF
3. Bone Marrow
4. Peripheral Blood
5. Fine Needle Aspirates
6. Lymph Node Biopsy

Specimen Acceptance Criteria:

1. Flow cytometry is not appropriate in any specimen for Hodgkin's disease. Flow cytometry is not appropriate in blood or bone marrow for anaplastic large cell lymphoma, lymphomatoid granulomatosis (LYG), thymic B cell lymphoma or large cell lymphoma.

2. Pre-approval required for all specimens: Contact Dr. Maryalice Stetler-Stevenson 301-402-1424 or Dr. Constance Yuan 301-496-4709.

3. Pre-scheduling required for all specimens: After approval, all specimens must be pre-scheduled via e-mail nciflowcytometry@mail.nih.gov or call the Flow Lab (301-402-1716) to schedule and to get bone marrow transport media if needed. Bone marrow transport media is for flow cytometry B1B58 specimens only.

4. A specimen cap is in effect . Even after approval, Emergency specimens will take priority over previously scheduled specimens. Emergency specimens are always processed. Only Dr. Stetler-Stevenson can designate a specimen as "Emergency" or "Rush".

5. Specimen labeling must include patient name and unique patient number . If the specimen is unlabeled or mislabeled, the clinician is required to identify specimen in person. Unlabeled or mislabeled specimens will not be processed.

6. All specimens must be ordered in CRIS prior to delivery . Enter Flow Cytometry orders under Anatomic Pathology as far in advance as possible (6 month advance order entry is not unusual for followup protocol specimens). The CRIS order printout in the Flow Cytometry department will be used as a requisition.

Note: CRIS order entry does not replace pre-approval or pre-scheduling. See #1 and #2, above. Specimens will not be accepted without an Active CRIS order. Processing may be initiated upon a verbal request to the pathologist (in #2, above) but processing will not be completed or reported without a requisition.

7. Phlebotomy Collection: Specimens to be collected by phlebotomy will require a second CRIS order. Enter the order as a Research Blood order and include specimen type, collection tube, volume, and STAT delivery to Building 10 Room B1B58 . If the nurse coordinator or fellow will collect and deliver the specimen directly to flow cytometry, the Research Blood order is not necessary.

8. Specimen rejection: Poor quality specimens are rejected unless the flow cytometry laboratory director approves processing for specimens that are rare; acquired at significant risk or suffering to the patient; or collection that cannot be repeated (i.e. a spleen can not be removed again).

   1. Specimen is examined grossly for clots, hemolysis and other gross characteristics. Viability assessment is performed for most specimens. Criteria for flagging specimen include: clotted specimen due to inadequate heparin, large volumes of bone marrow aspirate indicating heavy peripheral blood contamination, low viability due to excessive age of specimen or poor handling, coagulated or frozen specimen indicating not maintained at appropriate temperature, cell number too low, radioactive specimen, and hemolyzed specimen. These specimens are only processed if cleared by the laboratory director.

   2. Cells derived from blood, bone marrow, fluids and tissue must be viable for flow cytometry testing. Any specimen with fixative added for any length of time i.e. formalin, B5 fixative, alcohol, etc. will be rejected for flow cytometry testing. Fixation is not reversible. In addition, lithium heparin should not be used as it is toxic to cells.

   3. If no cells detected or less than 60% viability, Hematopathology and Cytopathology specimens are returned to these services. For other clotted or non-viable specimens, tests may be performed at discretion of the medical director if specimen is rare, acquired at significant risk or suffering to patient or can not be repeated. Results will be interpreted with caution.

   Emergency, Weekend and Evening Specimens:

• Contact the Pathology resident on call.
• Weekend and evening specimens will only be processed in extreme cases where there is a life threatening clinical situation requiring immediate treatment that will be based, at least partially, on Flow Cytometry results.
• Routine weekend and evening specimens can be sent to local reference laboratories at the expense of the clinical protocol.

Fluids Specimens:

1. Specimens go to Cytopathology. Note on the requisition that an aliquot should be sent to Flow Cytometry room B1B58.

CSF Specimens:

1. Because of the small number of cells present, only a limited panel can be set up.

2. CSF should be delivered ASAP as cells rapidly disintegrate and are not usable for flow cytometry after 3 hours. Specimen will be spun down, so supernatant could be used for other tests. Note accurate time of draw on the requisition or specimen may be rejected.

3. If specimens require a morphological diagnosis, they should be sent to Cytopathology with a note on the requisition that an aliquot should be sent to Flow Cytometry.

4. Deliver flow specimen and requisition immediately to the Flow Cytometry Laboratory, Building 10 Room B1B58.

5. If specimen is drawn because of patient neurologic changes, inform the flow cytometry lab and the specimen will be processed as a stat order.

Bone Marrow Specimens:

1. Notify Hematology that flow immunophenotyping is being performed. They will prepare an extra smear for the Flow Cytometry Laboratory when possible. This is in addition to scheduling with the Flow Lab (in 2 above) and entering a CRIS order (in 5 above).

2. Pick up bone marrow transport media from the Flow Cytometry Lab B1B58. Alternately, make arrangements to pickup transport media up to 24 hours in advance by calling 301-402-1716. Transport media is for B1B58 specimens only.

3. Get sterile heparin suitable for injection from the nurse's station.

4. Rinse syringe and needle with sterile heparin, leaving no more than 0.2-0.5 mL in syringe.

5. After aspirate for morphology, reposition needle and, for cellular specimens, slowly aspirate 2-3 mL of bone marrow. Aspiration of greater than 3mL results in significant dilution of the specimen in peripheral blood. For specimens with low cellularity, reposition and aspirate 3-5mL of bone marrow a second time. Note this on the requisition or the specimen will be rejected.

6. Immediately discharge aspirate into transport media, cap tube tightly and mix by gentle inversion 5-6 times. Label tube with patient name, unique identifier number and date.

7. The Active CRIS order should include the fellow's name and pager number, the patient history and clinical question. This information is vital as to the set-up strategy used. If rapid diagnosis and notification of result is required to begin treatment, indicate STAT nature, explain reason and leave name as well as pager number of fellow on evening call.

8. Deliver immediately to the Flow Cytometry Laboratory B1B58 (specimens containing hematopoietic neoplasms have a tendency to clot and must be processed immediately).

Peripheral Blood Specimens:

1. Draw at least 10mL blood into sodium heparinized (green) tubes. If the WBC is low, draw extra blood.

2. Deliver specimen and requisition immediately to the Flow Cytometry Laboratory, Building 10 Room B1B58.

Fine Needle Aspirates Specimens:

1. Specimens go to Cytopathology. Note on the requisition that an aliquot should be sent to Flow Cytometry room B1B58. Note specimen site on the requisition.

Lymph Node Biopsy Specimens:

1. Specimens go to Hematopathology. Note on the requisition that an aliquot be sent to Flow Cytometry room B1B58. Note specimen site on the requisition.

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Hematopathology Section Specimen Collection Manual

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

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Pediatric Tumor Biology & Ultrastructural Pathology

Specimen Submission for Electron Microscopy (EM) Exam

• Label specimen container with patient name and the submitting physician.  Fill out a request form and place it with the specimen container.
• Place fresh specimens for EM examination in glutaraldehyde fixative.  Buffered glutaraldehyde fixative is available from our lab upon request.
• Formalin-fixed specimens and paraffin-embedded tissues can also be submitted.  Place tissue from paraffin block into xylene before submission.
• Specimens can be mailed to: Section of Ultrastructural Pathology, Bldg. 10, Rm. 2A10, NIH, Bethesda, MD  20892.
• For inquiries, please contact: Mones Abu-Asab, (301) 496-2164.

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Specimen Collection and Handling For The Molecular Diagnostics Unit of the Laboratory of Pathology/NCI

The Molecular Diagnostics offers PCR-based molecular testing primarily designed to assist in the diagnosis of lymphoproliferative disorders.

I. Submission of IN-HOUSE samples:  

Specimens delivered to Rm. 2N110 in Bldg. 10 will be accepted no later than 4:30 pm.  

When submitting patient samples for PCR assays, the clinician or requesting physician must inform the Molecular Diagnostics Laboratory prior to the submission. This notification can be accomplished by entering a CRIS order at least 24 hr prior to submission of the sample. The CRIS order is listed under the heading of Anatomic Pathology, sub-heading, Molecular Pathology . The information submitted in the order must include the type of specimen and the requested tests.

When submitting patient samples for RT/PCR (for RNA), the clinician or requesting physician must inform the Molecular Diagnostics Laboratory prior to the submission. This notification can be accomplished either by faxing a completed "RT-PCR Request Sheet" 24 hr prior to submission of the sample (Fax number is: 301-402-2415) or by calling Dr. Lynn Sorbara (Technical Supervisor) at (301) 435 - 2627, or Ms. Tina Pham (Medical Technologist) at (301) 435-2632. The RT-PCR Request Sheet must be completed in order for the specimen to be accepted. Pertinent information regarding the clinical history, name, age, gender, and preliminary diagnosis as well as the name and phone number of the clinician must be included.

(1). BLOOD - 8 cc of whole blood is collected in "tiger topped" (Vacutainer CPT-Mononuclear Cell Preparation Tube by B-D) blue citrate tubes or regular light blue citrate tubes. The tubes are wrapped in plastic bags, (at room temperature) and sent immediately to the Molecular Diagnostics Unit.

(2). BONE MARROW ASPIRATES, CSF, FINE NEEDLE ASPIRATIONS, AND/OR OTHER TISSUES  

(a). Bone Marrow Aspirates - 1 cc of 1000 Units/ml of heparin is needed for each 2-3 ml sample of bone marrow aspirate. The syringe should be pre-treated with the heparin and 1 ml of the heparin must remain in the syringe with the sample. The syringe is capped and placed in a plastic bag (at room temperature). The bone marrow aspirate should be delivered immediately to the Molecular Diagnostics Unit.

(b). CSF - samples are placed in a sterile, capped plastic tube, wrapped in a plastic bag and then placed in a second bag or cup with wet ice. At no time should the ice and the tube have direct contact.  

If fresh CSF is not available, cytospins of the CSF that are, air-dried, and Diff-Quick stained may be submitted in lieu of fresh samples.

(c). Fine Needle Aspirations - all samples are placed into tubes with fresh, sterile RPMI media at room temperature. If fresh samples are not available, "smears" of the samples that are air-dried, and Diff-Quick may be submitted in lieu of fresh material.

(3). CT-GUIDED CORE BIOPSIES - For the submission of these samples, prior notification is essential. The Technical Supervisor will provide specific instructions for the preparation of these samples. Multiple services, such as, pediatrics, radiology, cytopathology, molecular, and surgical pathology need to be coordinated in their efforts.

A brief summary of the procedure - multiple passes of core biopsies are placed in a cryomould. The cores are then completely covered with liquid OCT and frozen in an alcohol/dry ice bath until completely solidified. The samples are kept frozen and delivered to the Molecular Diagnostics Unit immediately.

II. Submission of Samples from OUTSIDE of the NIH:

Notification of submission of samples from outside the NIH must to be made as a consultation to the Laboratory of Pathology. The Molecular Diagnostics Unit is not a reference laboratory.

(1). BLOOD - 8 cc of whole blood is collected in "tiger-topped" (Vacutainer CPT-Mononuclear Cell Preparation Tube by B-D) blue citrate tubes or regular light blue citrate tubes. The tubes are wrapped in plastic bags and "bubble wrap" to prevent breakage. The wrapped tubes are then placed into a Styrofoam box with "cold packs" (i.e., "picnic-type" frozen ice packs) and transported overnight by courier to the address given below.

(2). BONE MARROW ASPIRATES - 1 cc of 1000 Units/ml of heparin is needed for each 2-3 ml sample of bone marrow aspirate. The syringe should be pre-treated with the heparin and 1 ml of the heparin must remain in the syringe with the sample. The sample is transferred to a plastic sterile tube, wrapped in a plastic bag and "bubble wrap," placed into a Styrofoam box with "cold packs" for shipping by overnight delivery by courier to the address given below.

(3). FROZEN TISSUES AND CORE BIOPSIES - Dry ice or liquid nitrogen are used to "snap" freeze tissues. (The tissues are placed in small polycarbonate tubes or wrapped in aluminum foil prior to snap freezing.) The tubes and/or foil are wrapped in parafilm or small plastic bags and placed into a second larger tube to prevent "crushing" of the tissue during transportation. For overnight shipping, 7-10 lbs of dry ice is recommended during winter months and 10-12 lbs during summer months. Ship to the address below using the quickest courier service available.

Samples are sent directly to:
Molecular Diagnostics Unit
Lab. of Pathology/NCI/NIH
9000 Rockville Pike
Bldg. 10/ Room 2N110
Bethesda, MD 20892

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SPECIMEN COLLECTION

CONTACT INFORMATION

Intra-Operative Consult (Frozen Section) Resident: 102-10375 (8:30 - 5PM)

Hot Seat Resident: 102-10358 (8:30 to 5:00 PM)

Pathology Resident On-Call (before 8:30 AM, after 5 PM, weekends, holidays): (301) 496-1211 (NIH Page Operator)

Surgical Pathology Office: (301) 496-2441

Clinical Laboratory Manager: (301) 594-9532

Tissue Procurement Nurse: (301) 402-3106 or page 102-11902

Hematopathology Laboratory (301) 496-1567

Hematopathology Office (301) 496-0183

Histology Laboratory (301) 496-3126

Frozen Section Laboratory (301) 496-1110

INTRAOPERATIVE CONSULTS (FROZEN SECTIONS)

Page the Intra-Operative Consult (Frozen Section) Resident: 102-10375 for assistance. The Hot Seat Resident may be paged at 102-10358, as a back-up. Please provide the patient information and Operating Room Number when calling.

Please page the Pathology Resident On-Call if it is after 5 PM or on weekends or holidays by calling the NIH Page Operator at (301) 496-1211.

FRESH SPECIMENS FROM OPERATING ROOM

All fresh specimens collected in the Operating Room should be delivered to the Frozen Section room (10/2C533) with the proper identification labels. Each specimen must be recorded in the log book at the time of drop-off. All paperwork should be placed with the specimen or with the specimen log book. All specimens must be placed in the locked refrigerator labeled "For Surgical Pathology Specimens."

NOTE: If specimen is place in refrigerator after 4 PM or on weekends and holidays, please page the Pathology Resident on-call, through the NIH page operator at (301) 496-1211. The Resident may instruct you to place the specimen in 10% formalin (see instructions below).

PREVENT SPECIMEN AUTOLYSIS:

To prevent autolysis of fresh samples (without fixative) or drying out too quickly, it is i mportant to keep them cold (2°C to 6°C).

It is usually best to place a fresh specimen directly in the specimen container and keep cool in the refrigerator as soon as it is collected. Exceptions are noted below.

-To prevent small samples, such as needle core biopsies from drying out too quickly, place in Teflon pad and wet with cold (2°C to 6°C) saline, (pH 7.0 -7.4).

-Small curettage collections may be wetted with a very small volume cold (2°C to 6°C) saline, (pH 7.0 -7.4). , if necessary, if there is no blood of body fluid present from the collection process.

- NEVER USE WATER OR WARM SALINE!

IF TRANSPORT OF FRESH SPECIMEN IS DELAYED, place the specimen container in a specimen bag (double zip lock bag) and place on ice. Do NOT allow the container or specimen to come in contact with ice, because as it melts, the water may enter the specimen container.

NOTES:

1) For small biopsies, such as those collected during endoscopy, cystoscopy, and GYN procedures, please place specimens in 10% formalin fixative as soon as possible and affix a “ 10% formalin warning label”. For questions about fixation, please see below or page the HOT SEAT resident.

2) For Hematopathology specimens (lymph node biopsies) please contact the Hematopathology at (301) 496-1567 or the Hematopathology office at (301) 496-0183. Lymph node biopsies should be delivered to the Hematopathology Section directly for fresh tissue procurement.

DO NOT USE FIXATIVE ON LYMPH NODE SPECIMENS, UNLESS DIRECTED BY HEMATOPATHOLOGY.

For more information on Hematopathology specimens, please see: http://home.ccr.cancer.gov/LOP/Clinical/hematopath/hem_spec.asp

FIXED SPECIMENS

Specimens obtained in the clinics should be fixed in 10% formalin unless otherwise specified and brought to the Surgical Pathology Service, room 2A22 with a corresponding requisition form. Specimens without requisition forms will not be processed until proper identification of the specimen is received in the laboratory.

Containers should be properly identified with labels showing, name, medical record number, clinic location, type of specimen & site of biopsy (e.g., skin biopsy, left upper thigh).

Containers with fixative should have a label identifying the fixative (10% formalin warning label).

The amount of Formalin should be at least TWICE the volume of the specimen, but preferably TWENTY times the volume.

SPECIMEN LABELS (ALL SPECIMENS)

• All specimens must be labeled with name of the patient, medical record number, date of birth, unit number or OR room #, description ( site description, specimen number). The information on the labels MUST match the paperwork and MIS order. Please PRINT the name of the submitting physician on the label.

All RADIOACTIVE OR INFECTIOUS SPECIMENS, should be properly labeled and bagged.

PAPERWORK

Operating Room Specimen Record must be delivered by 4 PM. If patient is still undergoing surgery, please submit specimen record ASAP, but no later than 9 AM on the morning after surgery.

Research Specimens: Request & Certification for Research Procurement of Human Biological Materials (NIH-2803-1) This form must be delivered with research tissue; see section below regarding Research Procurement.

REQUISITION FORMS (MIS ORDERS FOR A LL CLINICAL SPECIMENS)

MIS requisition forms will print in 2A22: These forms must have the following information:

Request number

Name of patient, Medical Record number, DOB, Age, Sex, Unit Number

Intitial diagnosis, Protocol number, Institute

Name of Physician & Name of Physician entering the request

Specimen & site of biopsy (e.g., skin biopsy, left upper thigh).

MIS orders must be placed by the Surgeon or Surgical Fellow as soon as possible, and no later than 5 PM on the day of surgery. If the MIS is down, you must submit a MIS Downtime Requisition for Tissue Exam/Cytology Exam. The form is available at here.

The specimen description in the MIS must match the Operating Room Specimen Record and the specimen label as accurately as possible.

SPECIMENS WILL NOT BE ACCEPTED OR PROCESSED IN THE LABORATORY UNLESS PROPERLY LABELLED AND WITH THE CORRESPONDING REQUISITION FORM (MIS ORDER). Specimens without a MIS order will not be processed until proper identification of the specimen is received in the laboratory. In urgent situation, the pathologist may place the specimen in fixative while waiting for the order. Unlabelled or mislabeled specimens will not be processed unless properly identified, resolved to the pathologist satisfaction, and documented in the CC Occurrence Reporting System.

ATTEMPTS WILL BE MADE INMEDIATELY TO CONTACT THE RESPONSIBLE PHYSICIAN AND/OR PRIMARY CARE DOCTOR. IN URGENT SITUATIONS, THE CHIEF OF SURGERY BRANCH AND/OR INSTITUTE DIRECTOR MAY BE NOTIFIED.

THE CHIEF OF THE SURGICAL PATHOLOGY SECTION WILL BE NOTIFIED INMEDIATELY.

RESEARCH PROCUREMENT

When a patient has been scheduled for a surgical procedure or a biopsy

during which specimens will be collected for research the research team

or designated clinical staff should initiate the form, "Request &

Certification for Research Procurement of Human Biological Materials"

(NIH-2803-1) and obtain the signature of the PI/AI.  For inpatients, the

form is placed in the inpatient chart.  For outpatients, the form can be

sent to the PACU on the day of and prior to the procedure taking place.

We know there will be times when proactive planning will not work and a

patient is taken to the procedure area without the required Form NIH

2803-1.  You can bring the completed form to the PACU or the OR Front

Desk and the staff will make sure it is delivered to the right hands.

Research material will not be released unless the Request &

Certification for Research Procurement of Human Biological Materials

(Medical Records form NIH-2803-1) is complete and signed. Refer to

Medical Administrative Series M01-2 for more details.

If the specimen is time sensitive the research team should call the resident before the procedure takes place to coordinate quick release. If a critical research specimen requires prompt transport to the lab, the pathologist can be notified prospectively to make appropriate arrangements.

Most specimens removed during a surgical procedure must be reviewed grossly by an NCI pathologist before being released for research to insure that sufficient diagnostic material required for the clinical care of the patient has been submitted for clinical pathology evaluation. This includes any specimens that are divided in the Operating Room, but does not include the following exceptions:

• Protocol driven needle core biopsies or other small biopsies.
• Samples transported to other CLIA certified labs for division and test reporting of clinical results (e.g., bone marrow biopsies, muscle biopsies. ocular specimens, if they are sent to a CLIA certified lab that issues a diagnostic report).
• Samples which are transported to areas for division by LP Pathologists outside the operating room (e.g., TIL specimens, UOB nephrectomy & prostate specimens, lymph node biopsies divided by Hematopathology).

The OR team will call the Hot Seat resident (page 102-10358) to review

the specimen in either the surgical suite or frozen section room.

Shortly thereafter, the OR staff will notify the person designated on

Form NIH 2803-1 when the specimen is procured and/or ready for release

and give the location for pick up.

• If the Hot Seat is not available, the intra-operative consult resident (page 102-10375) will serve as a back up in emergency situations and the on-call resident will assist after hours and holidays (NIH pager operator, 6-1211). 

• The OR should also tell the pathologist and research team the OR suite # and phone number from which it can be obtained, if necessary.        
• If the research team arrives before the pathologist, please be patient and do not take the specimen until the pathologist reviews and releases it. 
• In some cases, the pathologist will review the specimen in the OR, but most research specimens will exit the operating room suite via the Frozen Section Laboratory (room 10/2C533) and be reviewed in that location.       

• Exempt samples most likely will not be passed through the Frozen Section Lab, since the lab is usually locked when not in use.

• While the OR is staffed until 11 PM Monday thru Friday, the doors do lock after 6 PM.  If you are picking up a specimen after 6:00 p.m., please use the phone located outside the PACU to call the Main OR number (301-496-5646) to advise them of your arrival.

For more information see:

http://home.ccr.cancer.gov/LOP/Clinical/labres/hbm.asp

RESERVATION OF MATERIAL All specimens will be kept, properly fixed in formalin for a period of a month in the Gross Surgical Pathology room (2A22). Then the specimens will be moved to the Autopsy room where they will be preserved for another month before discarding them. Processed (paraffin embedded) from clinical specimens are not discarded. OTHER TYPES SPECIMENS FOR LABORATORY OF PATHOLOGY

Last Updated 4/24/2008 11:31:11 AM