Fluroscence Cytometry-sysmex

2
FLUORESCENCE FLOW CYTOMETRY The Sysmex and haematology analyzers incorporate the ingenious Fluorescence Flow Cytometry technology, which was successfully introduced with top-of-the-end Sysmex . These X-series analyzers are equipped with excellent capabilities offering optimal differentiation between normal cells and pathological cells. XT-series XS-series XE-2100 range Q. How does Fluorescence Flow Cytometry function in X-series analyzers? A. As depicted in the image of Optical System of X-series analyzers, it detects each cell from three angles. 1. Forward Scattered Light (Information on Cell size) 2. Side Scattered Light (Information on Internal Cell structure) 3 . Side Fluorescence Light (Information on RNA/DNA content) Q. What is the role of Side or Lateral Fluorescence Light? A.Fluorescence Light detects DNA/RNA Information of cells along with information about cell activities such as Duplication activity of the nucleus (High RNA) Cytoplasm activity (Protein synthesis etc) This information is useful for very good separation of mature and immature cells. Q. What are the benefits of Fluorescence Flow Cytometry? Q. What is FAD? A. The major benefit of Fluorescence Flow Cytometry to its users is improved detection of pathological cells like Immature Granulocyte Count, High Fluorescence Lymphocyte Count, Abnormal Lymphocyte Detection etc. The other advantage is improved quality of 5 part differential of WBC with 48 hours stability after blood collection and that also for high linear ranges (no dilution necessary). It has a stable and long life detector system requiring no calibration. A. FAD is an acronym for Fluorescent Activated Differentiation used by X-series analyzers to do 5 part differentiation of WBC using Fluorescence Flow Cytometry. FREQUENTLY ASKED QUESTIONS Q. What is source of Fluorescence Light? A. Sysmex uses patented Polymethyne dye to stain DNA/RNA of cells. The stained DNA/RNA of the cells scatters 633 nm laser light, which is measured at 90° as Side Fluorescence Light (SFL).

description

Flouroscence Flowcytometry related FAQ from SYSMEX for easy understanding

Transcript of Fluroscence Cytometry-sysmex

Page 1: Fluroscence Cytometry-sysmex

FLUORESCENCE

FLOW CYTOMETRY

The Sysmex and haematology analyzers incorporate the ingenious Fluorescence Flow Cytometry technology, which was successfully introduced with top-of-the-end Sysmex . These X-series analyzers are equipped with excellent capabilities offering optimal differentiation between normal cells and pathological cells.

XT-series XS-series

XE-2100 range

Q. How does Fluorescence Flow Cytometry function in X-series analyzers?

A. As depicted in the image of Optical System of X-series analyzers, it detects each cell from three angles.

1. Forward Scattered Light (Information on Cell size)

2. Side Scattered Light (Information on Internal Cell structure)

3 . Side Fluorescence Light (Information on RNA/DNA content)

Q. What is the role of Side or Lateral Fluorescence Light?

A.Fluorescence Light detects DNA/RNA Information of cells along with information about cell activities such as

Duplication activity of the nucleus (High RNA) Cytoplasm activity (Protein synthesis etc)

This information is useful for very good separation of mature and immature cells.

Q. What are the benefits of Fluorescence Flow Cytometry?

Q. What is FAD?

A. The major benefit of Fluorescence Flow Cytometry to its users is improved detection of pathological cells like Immature Granulocyte Count, High Fluorescence Lymphocyte Count, Abnormal Lymphocyte Detection etc. The other advantage is improved quality of 5 part differential of WBC with 48 hours stability after blood collection and that also for high linear ranges (no dilution necessary). It has a stable and long life detector system requiring no calibration.

A. FAD is an acronym for Fluorescent Activated Differentiation used by X-series analyzers to do 5 part differentiation of WBC using Fluorescence Flow Cytometry.

FREQUENTLY ASKED QUESTIONS

Q. What is source of Fluorescence Light?A. Sysmex uses patented Polymethyne dye to stain DNA/RNA of cells. The stained DNA/RNA of the cells

scatters 633 nm laser light, which is measured at 90° as Side Fluorescence Light (SFL).

Page 2: Fluroscence Cytometry-sysmex

Q. How Polymethyne dye is useful in differentiation of cells?

A. Polymethyne dye allows the use of laser diodes which are significantly economical and more reliable than Argon ion lasers. The effect of staining of Polymethyne dye on mature and Immature cells is depicted in following illustrations.

Q. How does a scattergram of normal sample and abnormal sample look like?

A. As shown in the illustrations of scattergrams, DIFF channel uses Fluorescent Activated Differentiation (FAD); Side Fluorescence Light (Y-axis) and Internal Cell structure i.e. Side Scatter Side Scatter Light (X axis).

Mature cells before staining

Mature cells after staining

Immature cells before staining

Immature cells after staining

Due to higher DNA/RNA content, immature cells scatter more Fluorescence Light than mature cells.

FAD enables X-series analyzer to differentiate not only mature WBCs, but also mature and immature WBCs. By staining the cells analytical sensitivity of cell counter is enhanced and signal-to-noise ratio is increased. Reportable ranges are extended and interferences are reduced.

Normal Sample

LYMPH

EO

RBC -Ghost

NEUT & BASO

MONO

DIFF

Abnormal Sample

DIFF

Myelo

Blast

Atypical Lymph

Band

Promyelo

Metamyelo

Q. How scattergram is generated from Fluorescence Light?

A. The scattergram shown explains how scattergram is generated from Fluorescence (SFL) and Side Scattered Light (SSL) signals. Cells rich with RNA (high SFL) but with simple structure (low SSL) will be differentiated in population marked as 1). Cells poor with RNA (low SFL) but with simple structure (low SSL) will be differentiated in population marked as 2). Cells rich with RNA (high SFL) but with complex structure (high SSL) will be differentiated in population marked as 3).Cells poor with RNA (low SFL) but with complex structure (high SSL) will be differentiated in population marked as 4).

NRBC

PLT Clumps