Exam 1 - Canine Lymphoma Flow Cytometry Flashcards
how must samples be presented for flow cytometry in canine lymphoma?
in suspension!! blood, body fluids, & solid tissues that can be processed into single cell suspension
why is flow cytometry used for canine lymphoma?
immunophenotyping & to look at the DNA content of cells
T/F: flow cytometry is highly sensitive & specific & qualitative & quantitative
true
what are the 3 basic principles of the technology used in flow cytometry?
- cells in single file are interrogated by light
- light scatter & emission signals are captured
- specimens suitable for flow have to be sampled as a fluid or transformed into a cell suspension
how does flow cytometry work?
capture light scatter properties & light emitted by individual cells that innately fluoresce or that are labeled with fluorescent antibodies & then exposed to light
what 3 components can be measured with flow cytometry?
- forward scatter light signals
- side scatter light signals
- fluorescence emission signals
how is forward scatter light signal measured?
light that is refracted by a cell in the forward direction & continues in the same direction that the light was already traveling
what is forward scatter used for measuring?
cell size
what has a stronger forward scatter, a small cell or a big cell?
large cells
how is side scatter light signal measured?
light that is refracted by cells & travels in a different direction than its original path
what is side scatter light signals used for?
determining cell granularity & complexity of the cell
what cells have a higher side scatter?
highly granular cells with a high degree of internal complexity (neutrophils)
how is fluorescence emission signal measured?
fluorophore is used to stain for a protein of interest - most common are fluorescently conjugated antibodies - emit light after excitation with the laser
each dye or label has its own excitation & emission spectrum & creates a pulse of photon activity over time (‘peak’)
what components of antibody staining are expressed by surface antigens in fluorescence emission signals?
cell type, biological function, & development stage of the cell
what are the sample requirements for flow cytometry?
sufficient cells, free of clots & contaminants, fresh (48 hours max storage time), & samples from neoplasia with high cell turnover should be analyzed within 24 hours
what are 4 general ways for how veterinarians use flow cytometry?
- immunophenotyping lymphoma (b vs. t cell)
- more specific classification of different lymphoproliferative diseases which will impact treatment & prognosis
- classification of myeloproliferative diseases (acute leukemia)
- monitoring treatment response & disease status
chronic leukemia in dogs is most commonly what cell type?
most often T cell
what are the 2 t cell classifications of lymphoma in the dog?
large t cell & small t cell
what are the 2 lymphoma types seen in dogs?
t cell & b cell
abnormal hematopoiesis in the common myeloid progenitor cell leads to what kind of diseases?
myeloproliferative disease
abnormal hematopoiesis in the common lymphoid progenitor cell leads to what kind of diseases?
lymphoproliferative disease
why can stage 5 lymphoma patients be hard to differentiate from leukemia patients?
stage 5 lymphoma can have bone marrow involvement & acute or chronic primary leukemia can also have bone marrow involvement
what is the prognosis for acute lymphoid leukemia?
very poor
what is the prognosis for chronic lymphoid leukemia?
excellent
T/F: chronic lymphoid leukemia is often an incidental finding
true
what is the treatment for chronic lymphoid leukemia?
low dose oral chemotherapy
what are the clinical signs seen in chronic lymphoid leukemia?
none - typically asymptomatic
what is seen on cytology of chronic lymphoid leukemia?
circulating lymphocytes are usually small
what is seen on cytology of acute lymphoid leukemia?
intermediate to large in size circulating lymphocytes
what are the clinical signs seen with acute lymphoid leukemia?
usually sick animals - pyrexia, lethargy, & anorexia, moderate to marked cytopenias, mild to moderate enlargement of lymph nodes, spleen, or liver, & may have soft tissue involvement
what is the treatment for acute lymphoid leukemia?
aggressive supportive care & multi-agent chemotherapy protocol (CHOP)
T/F: myeloproliferative disease is rare in veterinary medicine
true
