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: CSF + 1 cytospin slide (if cytocentrifuge  is available)
Container/Tube:
Preferred from Referring Sites: Concentrated CSF 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.
Specimen Volume: 5-10 mL requested  (minimum volume 2 mL)
Collection Instructions:
1.  Collect specimen in a sterile vial.  Mix specimen well before taking an aliquot for flow cytometry testing.  CSF contaminated with peripheral blood is undesirable.  
2. For optimum results, specimen must be processed by the Flow Cytometry Lab immediately after collection. If specimen will not be processed by the Flow Cytometry lab immediately after collection, concentrate CSF sample and place in enhanced RPMI Flow Cytometry Transport Medium. Specimens should be prepared for transport as soon as possible after collection, preferably within 1 hour.
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.  Specimen cannot be frozen.
4.  Add-on testing to refrigerated CSF specimens is not possible due to cell loss.  
To concentrate or prepare a CSF specimen for transport in enhanced RPMI Flow Cytometry Transport Medium:
1. If a cytocentrifuge is available, prepare a cytospin slide from unpreserved CSF (use 10-15 drops CSF) specimen and send it with the concentrated specimen.
a.    Label cytospin slide with 2 accurate patient identifiers.  Handwrite the slide labels.  Do not use labels/stickers as patient identifiers on the slide.
b.    Do not stain slide.
2. Obtain a vial of enhanced RPMI Flow Cytometry Transport Medium (stored in a gray UV-blocking bag in the refrigerator).  
3. 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.
4. Place 5 to 10 mL of CSF in a large test tube suitable for centrifugation.
5. Centrifuge the fluid for 7 minutes at 2,000 RPM to form a cell pellet in the bottom of the tube.
6. Carefully remove the CSF from the centrifuge. Do not disturb the cell pellet in the bottom of the tube.
Note: You may not be able to see the cell pellet, but it is present.
7. Using a pipette, remove the supernatant from the tube. Leave the bottom 1 mL of fluid (contains the cell pellet) in the tube.
8. Vortex gently to resuspend the cell pellet in the remaining 1 mL of fluid.
9.  Add 1 mL of concentrated CSF to the vial of enhanced RPMI Flow Cytometry Transport Medium.
8. Mix by inverting the vial 3 times.
9. 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.
10. Place the sample in the gray UV-blocking biohazard bag for transport.
12. If a cytospin slide was prepared from the CSF specimen, send slide in a slide container in the same biohazard bag as the sample.
West Market laboratories please forward slides to Fargo laboratory for pathology interpretation.
 Additional information:  
For questions about sample requirements, please call the Flow Cytometry Laboratory at (218)786-5741 for more information (hours of operation 0630 – 1500).