phycoerythrin 4)85 are the most common fluorescent dyes used in the biomedical sciences. ight andor fluorescence scatter signals are detected by a series of photodiodes and amplified. :ptical filters are essential to bloc+ unwanted light and permit light of the desired wavelength to reach the photodetector. The resulting electrical pulses are digiti%ed$ and the data is stored$ analy%ed$ and displayed through a computer system.
The end result is quantitative information about every cell analy%ed 4Fig. 15. ince large numbers of cells are analy%ed in a short period of time 4<1$000sec5$ statistically valid information about cell populations is quic+ly obtained.
Flow Cytometry And Its Clinical Applications
Flow cytometry measures optical and fluorescence characteristics of single cells (or any other particle, including nuclei, microorganisms, chromosome preparations, and latex beads). Physical properties, such as size represented by forward angle light scatter) and internal complexity(represented by right-angle scatter) can resolve certain cell populations. Fluorescent dyes may bind or intercalate with different cellular components such as !" or #!". "dditionally, antibodies con$ugated to fluorescent dyes can bind specific proteins on cell membranes or inside cells. %hen labeled cells are passed by a light source, thefluorescent molecules are excited to a higher energy state. &pon returning to their resting states, the fluorochromes emit light energy at higher wavelengths. 'he use of multiple fluorochromes, each with similar excitation wavelengths and different emission wavelengths (or colors), allows several cell properties to be measured simultaneously.
Immunophenotyping of Leukemias and Lymphomas
*mmunophenotyping by flow cytometry is an important tool in the diagnosis and staging of patients with a haematological neoplasm. *t is used in con$unction with classical morphology.*n the bone marrow, normal blood cells develop from stem cells in a progressive series of differentiations, branching off to give different lineages of cells (for example, myeloid or lymphoid, ' cell or + cell), each of which has a distinct maturation pathway. "t each stage, the cells carry a distinctive set of marers classified by the (cluster of differentiation) nomenclature.'he panel selected will depend on initial clinical diagnosis. 'he results from the initial screen may indicate the need for further classification with an extended panel that is more specific for a particular disease. 'he marers include+ cell /, 01, 02, 31, 4/, 5appa, 6ambda7 ' cell 3, 8, 4, /, 9, :, 4/, /;7 <yeloid cells 9, 00b, 08, 04, 0/, 0;, 88, 84, 4/, /;, 009, =6"-#, <P>.Plasma cell 02, 8:, 4/, /;, 08:.
Detection of minimal residual disease
<inimal residual disease (<#) was defined as disease beyond the limit of morphological detection using conventional microscopy. Patients with acute leuaemia were considered to be in remission when bone marrow samples contained ?/@ neoplastic cells. Flow cytometric methods can detect far lower levels of disease, which can be important in the clinical management of leuaemia. 'he residual tumour cells are detected using immunofluorescence of surface marers.