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Specimen Collection Guide

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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Chromosome Pathology Instructions for Specimen Submission
Download FFPE-FISH Downtime Request Form

  1. In-house Cases
    For specimens collected at the NIH Clinical Center, requesting physicians will enter the request for FFPE FISH in the Clinical Research Information System (CRIS) in the same order used for requesting Surgical Pathology services. The box labeled “Tissue FISH Requested” should be checked and the specific tests should be listed in the ‘Special Instructions’ section at the bottom of the order.

  2. Outside (Submitted) Cases
    If FISH testing is needed on patient blocks or slides submitted from an outside facility for enrollment to a NIH protocol, a CRIS order for “Surgical Pathology – Outside Material”, should be entered. The Tissue FISH Requested box should be checked and specific tests entered into the “Special Instructions” box as for specimens collected at NIH. If a CRIS order cannot be entered because the patient is in pre-admit statues, fill out an “Outside Tissue Examination Form” (available in the Surgical Pathology Office). This form must be filled out completely and the specific FISH tests listed. Submit the CRIS order or Outside Tissue Examination Form along with the slides/blocks and the outside laboratory’s pathology reports. Please include the Outside Case Number, Block ID, and Tissue Source on your request.

  3. Iformation about tissue quality and submission of slides/blocks from other hospitals
    The amount of tissue required for molecular tests on solid tumors is variable and depends upon both specimen size and tumor content of the specimen. When you are requesting specimens from outside hospitals for evaluation at the NIH, a tissue block is preferred, for then we can control how the slides are prepared. However, many institutions will not release paraffin blocks and unstained slides must be requested instead. Please follow the guidelines below for specimen collection and handling:

  • Optimally, tissues should be fixed in formalin for 6-48 hours prior to processing and embedding. Tissue from B-5 or other mercuric fixatives generally does not hybridize well and will not be accepted.
  • Unstained sections should be cut at 5 micron thickness and mounted on TrueBond 380 (preferably) or positively charged slides.
  • For brain sections, two separate sections on a single slide are preferred.
  • Slide should be air (not oven) dried.
  • Slides must be clearly marked with the patient’s IDs.
  • Slides must be tested within 6 weeks of microtomy.
  • Store blocks and slides at room temperature (20-23.5°C) until submission.
  • Submit 4 unstained slides and 1 H&E slide (or 5 unstained slides) for each test.
 

Contact Information

Svetlana Pack, Ph.D
Head, Chromosome Pathology Unit,
Laboratory of Pathology, CCR, NCI, NIH
10 Center Dr., RM 2N115
Bethesda, MD 20892
Phone (301) 451-2723
Fax (301) 480-9488
Email

Zied Abdullaev, Ph.D
Molecular Biologist
Chromosome Pathology Unit,
Laboratory of Pathology, CCR, NCI, NIH
10 Center Dr., RM 2N115
Bethesda, MD 20892
Phone (301) 451-2711
Email

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Flow Cytometry Procedure Manual (Specimen Collections)

Flow Cytometry Specimen Scheduling and Delivery:

  1. Flow Cytometry is not appropriatein 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 is required for all specimens: Contact Dr. Maryalice Stetler-Stevenson or Dr. Constance Yuan.

  3. Scheduling is required for all specimens: After pathologist pre-approval, all specimens must be scheduled with the Flow Lab (301-402-1716).

  4. 1 p.m. Specimen Delivery cutoff time, Monday - Friday: Schedule Flow Cytometry specimen collection for first morning collection and Escort STAT pickup and delivery to B1-B58
    Flow Cytometry Lab, Room B1 B58 Specimens received before 1 p.m. will be processed the same day. Specimens arriving after 1 p.m. will not be processed without pathologist pre-approval. For late processing approval, contact Dr. Maryalice Stetler-Stevenson or Dr. Constance Yuan.

  5. 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".

  6. Emergency, Weekend, Holiday and Evening specimen delivery: Blood specimens collected 24 hours before a workday will be processed the next day i.e. on Sunday before a Monday or on a holiday before a workday. Deliver to the after hours drop box outside B1 B58. Blood specimens collected more than 24 hours before a workday will not be processed.

    For weekend or holiday delivery or processing of CSF, Fluid, or BM specimens, contact the Pathology Resident On-Call: (301) 496-1211 (NIH Page Operator). Weekend and evening specimens will only be processed in extreme cases when a life threatening clinical situation requires immediate treatment based, at least partially, on Flow Cytometry results. Send routine weekend and evening specimens to local reference laboratories billed to the clinical protocol.

Flow Cytometry Specimen Ordering and Labeling:

No specimen will be accepted without an Active CRIS order prior to delivery .
Under Order Entry, go to Anatomic Pathology and choose either “Flow Cytometry CSF” or “Flow Cytometry Order Set” (see specimen type 1 - 4, below). Complete the requested information. Include the fellow’s name and pager number, patient history and clinical question.

Enter Flow Cytometry orders for all follow up orders as far in advance as possible (6 month advance order entry is not unusual for follow up protocol specimens).

STAT results: If rapid diagnosis and notification of result is required to begin treatment, indicate STAT nature in the CRIS order Special Instructions section. Include name and pager number of fellow on evening call.

CRIS order entry does not replace pre-approval or pre-scheduling. Specimen processing may be initiated upon a verbal request to the pathologist, but processing will not be completed or reported without a CRIS order.

Specimen labeling must include patient name and unique patient number. If the specimen is unlabeled or mislabeled, the clinician is required to identify the specimen in person. Unlabeled or mislabeled specimens will not be processed.
  1. CSF: In CRIS under Order Entry, select Anatomic Pathology / Flow Cytometry CSF. Complete the requested information.

  2. Peripheral Blood: In CRIS, select Anatomic Pathology / Flow Cytometry Order Set. Complete the Flow Cytometry order requested information.

    Complete the Research Blood order by adding this information: Peripheral blood, 20ml sodium heparin green top tube. Deliver STAT to Flow Cytometry Room B1-B58. For specimens shipped overnight, deselect Research Blood order.

  3. Bone Marrow: In CRIS, select Anatomic Pathology / Flow Cytometry Order Set. Deselect the Research Blood order. Complete the Flow Cytometry order requested information.

  4. Non-CSF Fluids, non-BM biopsies and aspirates (LN/Tissue/FNA) : Enter two Anatomic Pathology orders: Cytopathology and Flow Cytometry Order Set. Fluids, FNAs and biopsies require triage through Cytopathology. Note on the Cytopathology CRIS order an aliquot should be sent to Flow Cytometry.

    Deselect the Research Blood order in the Flow Cytometry Order Set. Complete the Flow Cytometry order requested information including source / location prior to delivery to Cytopathology.

Flow Cytometry Specimen Transport Media and Delivery:

Specimen delivery cutoff time is 1 p.m. Monday – Friday workdays (no weekends or holidays).
Deliver blood, bone marrow, CSF, FNAs, fluid or other cell suspensions at ambient temperature (20-25C). Deliver frozen cells on dry ice.

  1. Peripheral Blood
    -- requires 10 - 20ml blood in sodium heparin green top vacutainer tubes. For Absolute Lymph # less than 1.0 K / uL, collect an additional 10ml. Deliver STAT at ambient temperature (room temperature, no cold packs).
    -- Call for STAT Escort pickup and delivery (301-496-9295) to Flow Cytometry B1 B58 if you cannot deliver the blood specimen yourself.

  2. Bone Marrow aspirate
    Notify the CCR Hematology lab that Flow Cytometry immunophenotyping is being performed (301-496-4473). The hematology bm collection tech will bring a 10ml sodium heparin green top vacutainer tube to the specimen collection site and prepare an extra smear for the Flow Cytometry Laboratory.

    Bone Marrow Collection IMPORTANT!! The fellow or assisting nurse must dispense 3 - 5ml BM aspirate IMMEDIATELY into the BM transport tube, cap the tube and invert 5 times to mix well. Label with patient information and deliver STAT to B1B58 at ambient temperature (room temperature, no cold packs)

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

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

    3. After aspirate for morphology, reposition needle and slowly aspirate 3-5 mL of bone marrow for flow cytometry.

    4. Immediately discharge syringe into open sodium heparin green top tube, cap tube and mix by gentle inversion 5-6 times.

    5. Label tube with patient name, unique identifier number and date.

      Call for STAT Escort pickup and delivery (301-496-9295) to Flow Cytometry B1 B58 or deliver STAT to room B1 B58 yourself.

  3. Cerebral Spinal Fluid (CSF)
    Rrequires a CSF Transport media tube. Before collection, pick up a transport media tube from CCR 3SW North Procedure UnitEndoscopy suite (3-3613 Utility room refrigerator) or from the Flow Lab B1-B58 or call the flow lab (301-402-1716).

    CSF collection: The fellow or assisting nurse must dispense 3-4ml CSF IMMEDIATELY into the CSF transport tube, cap the tube and invert 5 times to mix well. Label the tube with patient information. Deliver STAT to Flow Cytometry B1 B58.

    Call for STAT Escort pickup and delivery (301-496-9295) on ice to Flow Cytometry B1 B58 if you cannot deliver CSF specimen yourself.

  4. Non-CSF Fluid, Fine Needle Aspirate or LN / Tissue Biopsy
    All fluids, FNAs and biopsies require triage through Cytopathology. Note on the Cytopathology CRIS order an aliquot should be sent to Flow Cytometry.
    IMPORTANT!! Do not add any fixative to the Flow Cytometry aliquot i.e. formalin, B5 fixative, alcohol, etc. Fixation will cause cell death and is not reversible.

Flow Cytometry Specimen Rejection:

Specimens arriving after 1 p.m. will not be processed without pathologist pre-approval. Contact Dr. Maryalice Stetler-Stevenson 301-402-1424 or Dr. Constance Yuan 301-496-4709 for late processing approval.

Unlabeled or mislabeled specimens will not be processed unless the clinician or collecting personnel can identify the specimen in person.

Poor quality specimens will be rejected (see examples below); poor quality specimen processing is at the discretion of the Flow Cytometry laboratory director. The director may approve processing of rare specimens; specimens collected at significant risk or suffering to the patient; or specimens that cannot be recollected i.e. a spleen can not be removed again. Results of poor quality specimens will be interpreted with caution.

The Flow Cytometry laboratory director or alternate will contact the clinician for recollection of rejected specimens. The rejected specimen will be accessioned and finalized. In addition, the clinician will be notified and the specimen will be held at room temperature for retrieval by the sending service i.e. Hematopathology, Cytopathology or clinical group for alternate testing. Specimens remaining in the Flow Cytometry lab will be discarded the following work week.
Poor quality specimens include:

    1. Damage or improper handling detected by gross examination of Specimen: Specimen clotting may be due to inadequate heparin. Coagulated, hemolyzed or frozen specimen may indicate non-ambient temperature conditions. Inappropriate collection containers include lithium heparin tubes or other non-approved anticoagulants. Specimens labeled radioactive or tuberculin hazard are rejected due to unavailable safety equipment in the testing area.

    2. Inappropriate specimen volume:
      Bone marrow aspirate volume greater than 5ml indicates probable peripheral blood contamination unless needle repositioning is noted. Specimens with very low cellular content may be rejected.

    3. Fixed Specimen / Specimen in Fixative:
      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.

    4. Low Viability / Non-Viable cells: Viability less than 60% may be rejected. Cells derived from blood, bone marrow, fluids and tissue must be viable for flow cytometry testing. Cell death may be due to excessive age of specimen, poor handling, or patient therapy.

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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

Contact Info: Tina Pham - 301-435-2632 / Trinh Pham 301-435-5234 / Winnifred Navarro 301-435-2627 / Liqiang Xi 301-594-1525

The Molecular Diagnostics Unit offers PCR-based molecular testing designed to assist in the diagnosis of both solid tumors and lymphoproliferative disorders.

I. Submission of IN-HOUSE samples:  

Specimens delivered to Rm. 2N116A in Bldg. 10 will be accepted no later than 4:30 pm.  

When submitting patient samples to the molecular diagnostics unit, the clinician or requesting physician must inform the Molecular Diagnostics Unit 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. Pertinent information regarding the clinical history, name, age, gender, and preliminary diagnosis as well as the name and phone number of the clinician should also be included.

(1). BLOOD - 5 cc of whole blood is collected in 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, AND OTHER TISSUES  

(a). Bone Marrow Aspirates -

  • Draw 2-3 cc in plain syringe with no anticoagulant. DO NOT USE HEPARINIZED SYRINGE

  • IMMEDIATELY place sample into the EDTA tube (lavender top) and mix gently to prevent clotting

(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.

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 - 5 cc of whole blood is collected in 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 -

  • Draw 2-3 cc in plain syringe with no anticoagulant. DO NOT USE HEPARINIZED SYRINGE

  • IMMEDIATELY place sample into the EDTA tube (lavender top) and mix gently to prevent clotting

The sample is 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. Do not freeze.

(3). Submitted FFPE tissues – All FFPE biopsy tissues performed for molecular diagnostic testing should be submitted routinely through surgical pathology. The surgical pathology staff is responsible for triaging the case for all specialized testing, including molecular diagnostic assays. It is essential that separate CRIS orders for both surgical pathology examination, and molecular diagnostic testing are submitted, and the required tests are listed on both CRIS forms. If the patient does not have an active MRN, then it is essential that a tissue examination sheet be completed in the Surgical Pathology main office (Building 10/2B50), and all required assays are listed on this form. Click the links to see specific detailed submission guidelines for Neuro-oncology and Thoracic Oncology .

Blood and bone marrow samples should be sent directly to: Molecular Diagnostics Unit, Lab of Pathology/NCI/NIH 9000 Rockville Pike, Bldg. 10/ Room 2N116A, Bethesda, MD. 20892

All other tissues samples should be brought to the main Surgical Pathology office (Building 10/2B50).

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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 CRIS 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 (CRIS ORDERS FOR ALL CLINICAL SPECIMENS)

  • CRIS 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).

    • CRIS 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 CRIS is down, you must submit a CRIS Downtime Requisition for Tissue Exam/Cytology Exam. The form is available at here.

    • The specimen description in the CRIS 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 (CRIS ORDER). Specimens without a CRIS 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:

    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 2/6/2012 4:22:50 PM

     
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