Using standard cellular biology techniques, researchers at UCI have developed a method for detecting the cellular components of blood easily, cheaply, and quickly with accurate quantification using fluorescence techniques.
The cellular components of blood (red blood cells, immune cells, platelets, etc.) offer information about the state of a person’s health and wellness. For sophisticated analysis of whole blood, researchers at labs and hospitals employ flow cytometry, a cell biology technique that separates cells based on their optical characteristics. A major challenge in analyzing whole blood by flow cytometry is that red cells far outnumber immune cells and rare cell types like stem cells or cancer cells, making the less numerous cells difficult to quantify, isolate, and analyze. Current solutions to capture these uncommon cells include the use of fluorescently-tagged antibodies, but this requires buying multiple expensive antibody reagents to separate multiple cell types.
Researchers at UCI have developed a method for the use of a single reagent, fluorescein diacetate, to successfully separate the cellular components of blood from complex biological mixtures. Fluorescein diacetate is not optically active until it enters a live cell, which converts it to the green fluorescent molecule fluorescein. Different blood cell types produce different amounts of fluorescein, which coupled with their unique forward scatter in flow cytometry allows for accurate quantification and isolation for further characterization. This method offers a cheaper, more straightforward solution or companion to the antibody-staining method for blood cell separation and identification of rare cell types associated with health and disease (T cells, lymphomas, peripheral stem cells, etc.)
·Utilizes flow cytometry, a standard cell biology technique
·Cheaper than antibody reagents
·Accurate quantification of cell types
·Staining with fluorescent probe is fast and reliable
|United States Of America||Issued Patent||6,696,241||02/24/2004||1996-371|
Proof of concept studies conducted in vitro, including: the identification of red cells, lymphocytes, monocytes, and granulocytes from whole blood; subset analysis on CD4- and CD8-positive T cells; identification of leukemias and lymphomas; and isolation of peripheral stem cells.