Pathology, Hematopathology Services

1. Bone marrow aspirate smears and/or Touch Preps, air-dried smears; unstained smears are preferred.

2. Bone marrow aspirate clot in 10% buffered formalin.

3. Bone marrow core biopsy in 10% buffered formalin.

4. Complete blood count (CBC) data with WBC differential. Results from recent testing, automated or manual testing is acceptable. If results are not available, draw and submit blood per instructions under CBC.

5. Peripheral blood smear, preferably corresponding to date of CBC.

Specimen Type: Bone marrow

Container/Tube: See #2 in Collection Instructions

Collection Instructions:

1. This procedure should be followed by a person who is skilled in the preparation of bone marrow particle smears.

2. Pre-assemble the following:

a. Using a #2 lead pencil, pre-label 6 slides with at least 2 sources of patient identification (eg, bar code, patient name, date of birth, or other unique identifier), and date of collection.

b. Standard size lavender-top (EDTA) tube.

c. Standard size green-top (heparin) tube.

d. Container with 10% neutral buffered formalin.

e. 10-mL syringe and needles

f. At least 2 additional patient labels.

3. Operator should pre-heparinize sterile syringe of appropriate size to aspirate at least 5 mL of bone marrow. There should be a dry (non-heparinized) sterile syringe available of appropriate size to aspirate at least 2 mL of bone marrow.

4. After appropriate site prep and local anesthesia, biopsy needle is first inserted, with trocar in place, through cortex of bone just into marrow space.

5. Trocar is removed and a dry (non-heparinized) syringe is attached. One mL to 2 mL of aspirate is withdrawn and syringe is immediately detached and handed to technical assistant.

6. Technical assistant will prepare particle smears from aspirate in the usual manner allowing unused material remaining in syringe to clot. Smears are rapidly air-dried (do not close folder while slides are still wet).

7. Allow remaining bone marrow aspirate in syringe to clot.

8. Place clot in container with enough 10% neutral buffered formalin to cover specimen.

9. Label container as bone marrow clot.

10. If technical assistant reports that no particles have been obtained, operator may elect to reposition needle and attempt another aspirate.

11. After technical assistant has finished preparing smears from initial aspirate, operator attaches second, previously-heparinized syringe to biopsy needle, and an aspirate of approximately 5 mL is withdrawn.

12. Syringe is immediately detached and contents divided between tubes as follows:

a. Flow cytometry - 1 mL of bone marrow in a green-top (heparin) tube (preferred) or a lavender-top (EDTA) tube

b. Molecular or cytogenetics studies - 1 full, green-top (heparin) tube

Note: Assistant removes stoppers and holds tubes as operator transfers bone marrow aspirate without use of a needle. Assistant then replaces stoppers and immediately mixes tubes by inverting multiple times.

13. Needle is then advanced into bone approximately 1 inch, after appropriate maneuvers withdraw needle.

14. Remove core from needle by inserting trocar into tip of needle and pushing core out through base.

15. Place core in container with enough 10% neutral buffered formalin to cover specimen.

16. Label container as bone marrow biopsy.

17. Label container with at least 2 sources of patient identification (eg, bar code, patient name, date of birth, or other unique identifier), date of collection, and type and site of specimen.

18. Forward promptly. If there is a delay in transport, all non-fixed specimens should be transported on wet-ice.

CBC

Specimen Type: Whole blood

Container/Tube: Lavender-top (EDTA) tube

Specimen Volume: Full tube (minimum: 4 mL or 250 µL for microtainer)

Transport Temperature:  Ambient

Collection Instructions: 

1. Invert several times to mix blood.

2. Clotted or hemolyzed specimen, specimen diluted with intravenous fluid, specimen less than minimum volume, or specimen ≥8 hours old at ambient temperature or ≥24 hours old at 4° C is not acceptable.

Tissue

Specimen in formalin fixative and fresh specimen (in saline or nutrient medium) are required.

Sources: Lymph node biopsies, spleen, or tumorous lesions suspected to be lymphoid or hematopoietic neoplasms

Specimen Type: Tissue in formalin

Container/Tube: Container with 10% neutral buffered formalin

Specimen Volume: ≥0.5 cc

Collection Instructions:

1. Submit tissue in container with enough formalin to cover specimen.

2. Orient tissue if appropriate.

3. Label container with at least 2 sources of patient identification (eg, bar code, patient name, date of birth, or other unique identifier), date of collection, and type of specimen.

Specimen Type: Tissue, fresh

Container/Tube: Container with saline or nutrient medium - no preservative

Specimen Volume: ≥0.5 cc

Collection Instructions:

1. Orient tissue if appropriate.

2. Label container with at least 2 sources of patient identification (eg, bar code, patient name, date of birth, or other unique identifier), date of collection, and type and site of specimen.

Note:

1. Complete a SaraPath Diagnostics requisition containing the following information:

a. Patient’s name and at least 1 other unique identifier (eg, bar code label or date of birth)

b. Name of clinician requesting procedure, operator performing procedure, and any other physicians who should receive a report by including phone and fax numbers

c. Appropriate clinical information including working diagnosis, questions of relevance, and special requests

2. Verify patient name and other unique identifiers on requisition match those on specimen.

3. Specimen may be rejected and returned to originating site if any of the following conditions apply:

a. Patient or specimen identification errors or omissions

b. Incorrect specimen submitted for test requested

c. Damaged or leaking specimen

d. Irreparable slide

4. Call SaraPath Diagnostics for:

a. Technical assistance - Monday through Friday, 8 a.m. to 5 p.m. at 941-362-8900

b. Courier service or supplies - Monday through Friday, 8 a.m. to 4 p.m. at 941-362-8932 or 941-362-8900

c. Schedule frozen section - Monday through Friday, 8 a.m. to 4 p.m. at 941-362-8932 or 941-362-8900

Note: STAT Specimen - Call for pick-up of STAT specimen and indicate STAT on requisition.