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Veterinary Pathology

Clinical Immunodiagnostic Laboratory

Veterinary Clinical Flow Cytometry 

Veterinary Clinical Flow Cytometry Facts [PDF]

What is flow cytometry? 

Flow cytometry is a test that determines the immunophenotype of cells by recognizing normal and abnormal expression of cell surface markers on T cells (helper T cells, cytotoxic T cells), B cells, neutrophils, and monocytes.  We offer clinical flow cytometry for cats and dogs.

Clinical flow cytometry is most often used: 

  1. To help distinguish between reactive and neoplastic lymphocytes
  2. To determine if lymphoma/leukemia is of B or T cell origin and in some cases, identify specific subtypes of B or T cell lymphoma/leukemia which will aid in prognosis
  3. To differentiate between lymphoid and non-lymphoid (myeloid/monocytic) neoplasia

Clinical flow cytometry is used most often in combination with clinical history/presentation and cytology/blood smear examination for diagnosis of disease. In some cases, additional diagnostics (PARR- PCR for antigen receptor rearrangement, histopathology, immunohistochemistry, immunocytochemistry) are also required. 

When might clinical flow cytometry be most helpful?

  1. Lymphadenopathy, organomegaly, or mediastinal mass with the following cytology results:
    1. Confirmed or probable lymphoma composed of intermediate to large lymphocytes
    2. Homogeneous expansion of small to intermediate lymphocytes
    3. Suspected lymphoma or thymoma (for mediastinal mass)
      Sample to submit: Fine needle aspirate of lymph node/enlarged organ/mass (see sample submission guidelines)
      Note: Flow cytometry is not indicated on these samples if cytology results report rare to low numbers of atypical/suspicious cells
  2. Abnormal CBC findings consisting of:
    1. Peripheral lymphocytosis with increased numbers of small mature, intermediate, or immature lymphocytes
    2. Presence of any immature cells/’blasts’ with a normal or elevated white blood cell count
      Sample to submit: Peripheral blood (EDTA tube)
      NOTE: If the lymphocytosis is composed of small to intermediate lymphocytes test for Ehrlichia infection in dogs with lymphocyte counts up to ~50k and ‘stress’ in cats with lymphocyte counts up to ~30k prior to running flow cytometry
  3. Abnormal bone marrow findings with the following cytology/CBC results:
    1. Increased numbers of blasts or small/intermediate lymphocytes with leukopenia on CBC
      Sample to submit: Bone marrow (EDTA tube) for samples with leukopenia on CBC. Peripheral blood (EDTA tube) is preferred if lymphocytosis is present.
  4. Body cavity effusion with the following cytology results:
    1. High numbers of small/intermediate/OR large lymphocytes consistent with or concerning for lymphoma
      Sample to submit: Cavity fluid (EDTA tube and red top tube when possible- 0.5ml in each tube, see sample submission guidelines)

Uncertain if flow cytometry will be helpful for your patient? Email vetclinflow@uga.edu or contact (706)542-9430 alt phone: (706)542-5161

Sample collection and submission guidelines

Clinical Flow Cytometry Submission Form [PDF]

Lymph node, organ or mediastinal mass aspirates

  • Put 1 ml of 0.9% saline in a red or white top tube (no additives, no serum separator)
  • Add 0.1ml of serum from the same patient or another patient of the same species
    • Note: the volume of serum should be ~10% of the total volume (this helps preserve cells)
  • Using a 22ga needle and 6 or 12cc syringe, aspirate with suction
  • Eject the contents into the saline, gently aspirate the saline into the syringe and eject back into the tube to recover all cells
  • Aspirate several times (2-3X if possible) repeating the above process until the saline is slightly cloudy

Blood and bone marrow

  • Provide a minimum of 0.5ml in an EDTA tube
  • A current CBC (within 2days) of the flow sample collection is required
    • Include a copy of your patient CBC
      OR
    • Check the CBC test request box and include: 1) a minimum of 2.0 ml of blood in an EDTA tube (use the appropriate fill volume for your tube) for the CBC and flow cytometry and 2) one unstained blood smear
      • See pricing schedule for CBC

Body cavity fluid

  • Provide a minimum of 0.5ml in an EDTA tube
  • If enough fluid is available please provide an additional red top tube (no additives, no serum separator) with a minimum of 0.5ml
  • If the total protein is < 5mg/dl please add two drops of serum from the same patient or another patient of the same species (if protein measurement is unavailable, add 2 drops of serum regardless)

Sample shipping guidelines

  • Collect samples and ship on the same day overnight for Monday through Friday delivery
  • Keep samples refrigerated (DO NOT FREEZE) until shipped
  • Ship overnight with an ice pack
  • Samples may be submitted via:

University of Georgia, College of Vet Med
Department of Pathology
Veterinary Clinical Flow Cytometry (Lab 155)
501 D.W. Brooks Drive
Athens, GA

Reporting and pricing schedule

  • Samples will processed on the day they are received or the next day to determine the cell count and cell viability and perform flow cytometry
  • Note: If a sample has too few cells or the cell viability is too low, flow cytometry results will not be accurate and flow cytometry will not be performed.
    • A fee of $15 will be charged for the initial sample assessment in these cases
  • Pricing Schedule
    Canine and Feline Flow Cytometry $115.00
    Cytology Review $35.00
    Body Cavity Fluid $40.00
    CBC $20.00
    Blood Smear Review $15.00
    AVDL Accession Fe $3.00
    AVDL Courier Services (Optional) $10.00 (For more information on courier servies, please click here.)
  • Reporting
    • Reports will be generated within approximately 3 business days of sample processing

Questions?

Email: vetclinflow@uga.edu
Phone: (706)542-9430
Alternate phone: (706)542-5161

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