Specimen Requirements

Specimen source, date of collection, and an order comment stating the reason for request including pertinent patient history is required at order entry.

Specimen Type: Tissue or fine needle core biopsy (FNA)

Container/Tube:

Preferred from Referring Sites: Tissue or fine needle core biopsy/FNA sample in enhanced RPMI Flow Cytometry Transport Medium

NOTE: The vials of enhanced RPMI Flow Cytometry Transport media are shipped to referring sites in gray UV-blocking biohazard bags to prolong the media shelf-life. The media is stored in the refrigerator.

Acceptable: Tissue or fine needle core biopsy/FNA in Hanks Balanced Salt Solution

Specimen Volume: Entire Specimen

Collection Instructions:

1. Specimens should be prepared for transport as soon as possible after collection, preferably within 1 hour. See directions below.

3. Forward promptly to Flow Cytometry Lab at ambient temperature. Specimens should be received in the Flow Cytometry Lab within 24 hours of collection or as soon as possible thereafter. It is recommended that testing be completed within 48 hours of specimen collection for optimal cellular viability.

4. Add-on flow cytometry testing to specimens that have been refrigerated is acceptable, however alterations in antigen expression may be seen following refrigeration. Specimens cannot be frozen.

5. Specimens placed in formalin are not acceptable for flow cytometry testing.

To prepare a tissue specimen for transport in enhanced RPMI media:

1. Check that the enhanced RPMI Flow Cytometry Transport Medium is salmon/pink in color and has an acceptable expiration date. Do not use the media if it is yellow in color.
2. Obtain a vial of enhanced RPMI Flow Cytometry Transport Medium (stored in a gray UV-blocking bag in the refrigerator).
3. Obtain a representative piece of tissue approximately the size of a number 2 pencil eraser. Use a scalpel and forceps to cut this piece of tissue into 5 to 6 smaller pieces.
4. Using a forceps, place the small pieces of tissue into the vial of enhanced RPMI Flow Cytometry Transport Medium.
   • Mix by inverting the vial 3 times or gently vortex the vial containing the tissue.
   • Label the vial with a patient label that includes:

a. The patient’s name

b. The patient’s medical record number

c. The flow cytometry order number (if applicable)

d. The date and time of specimen collection

7. Place the sample in the gray UV-blocking biohazard bag for transport.

To prepare a fine needle core biopsy/FNA specimen for transport in enhanced RPMI media:

Note: It is ok to transfer a piece of tissue to RPMI media without centrifuging. If fine needle core biopsy/FNA specimen is collected in a large amount of saline or Hanks Balanced Salt Solution (HBSS), or is bloody, then centrifuge.

1. Label the sample vial with a patient label that includes:
2. Mix by inverting the vial 3 times or gently vortex the vial containing the core needle biopsy/FNA.
3. Transfer the cell pellet into a vial of enhanced RPMI Flow Cytometry Transport Medium and mix gently to resuspend the cell pellet in the remaining 1 mL of fluid.
4. Using a pipette, remove the supernatant from the tube. Leave the bottom 1 mL of fluid (contains the cell pellet) in the tube.
5. Carefully remove the fluid from the centrifuge. Do not disturb the cell pellet in the bottom of the tube.
6. Centrifuge the fluid for 7 minutes at 2,000 RPM to form a cell pellet in the bottom of the tube.
7. Check that the enhanced RPMI Flow Cytometry Transport Medium is salmon/pink in color and has an acceptable expiration date. Do not use the media if it is yellow in color.
8. Obtain a vial of enhanced RPMI Flow Cytometry Transport Medium (stored in a gray UV-blocking bag in the refrigerator).

a. The patient’s name

b. The patient’s medical record number

c. The flow cytometry order number (if applicable)

d. The date and time of specimen collection

9. Place the sample in the gray UV-blocking biohazard bag for transport.