Flow cytometry blood analysis

Several additional instrumental techniques have also been developed for bacterial characterization. Capillary electrophoresis of bacteria, which requires little sample preparation,42 is possible because most bacteria act as colloidal particles in suspension and can be separated by their electrical charge. Capillary electrophoresis provides information that may be useful for identification. Flow cytometry also can be used to identify and separate individual cells in a mixture.11,42 Infrared spectroscopy has been used to characterize bacteria caught on transparent filters.113 Fourier-transform infrared (FTIR) spectroscopy, with linear discriminant analysis and artificial neural networks, has been adapted for identifying foodbome bacteria25,113 and pathogenic bacteria in the blood.5... 

Lombardi, V.R.M., Amado, L., Femandez-Novoa, L., Ftcheverrfa, 1., Seoane, S., Cacabelos, R. (2003) Flow cytometry analysis of CD28-/CD8-I- suppresor cell precursor and CD45RO-I-/ CD4-I- memory T lymphocytes in the peripheral blood of Alzheimer s disease patients. In New trends in Alzheimer- and Parkinson-related disorders, Hanin, 1., Fisher, A., Cacabelos, R. (eds.), Monduzzi Fditore, Bologna, pp. 57-61. 

Several methods for separating cells have been devised. These include electrophoresisd or use of magnetic microspheres. b Micromanipulation can sometimes be used to select single cells for analysis. The most impressive technique is flow cytometry,ef which is used daily on human blood samples in clinical laboratories. A suspension of cells is passed at a high rate of flow through a narrow capillary of 0.2 mM diameter. The sample stream, which is surrounded by a larger "sheath" stream, has a... 

Traill, K. N., Bock, G, Winter, U, Hilchenbach, M., Jurgens, G., and Wick, G. (1986) Simple method for comparing large numbers of flow-cytometry histograms exemplified by analysis of the CD4 (T4) antigen and LDL receptor on human peripheral-blood lymphocytes. J Histochem. Cytochem. 34, 1217-1221... 

The second aspect of clinical practice that has led to a reassessment of the nature of flow cytometry is the occasional clinical requirement for rare-event analysis. Methods have been developed, particularly with the use of multiparameter gating, to lower background noise in order to provide increased sensitivity for detection of rare cells. In the clinic, this increased sensitivity translates, for example, into earlier diagnosis of relapse in leukemia, more sensitive detection of fetal-maternal hemorrhage, and better ability to screen leukocyte-reduced blood transfusion products for residual white blood cells. Outside the clinic, these methods for rare-event detection have begun to stretch the limits of research applications as well. 

In sum, EMPs have emerged as a preferred direct method for assessing EC injury in different disorders. EMP analysis could provide insight into the actual status of the endothelium in vivo by a simple blood analysis. However, there is a need for refinement and standardization of the assay method. Overall, most groups have relied on flow cytometry for the measurement of EMPs nevertheless, other methods such as ELISA are available and may be an option in the future. The main challenge remains in the selection of specific and sensitive monoclonal antibodies that may yield consistent results between different laboratories. In addition, the protocols for sample handling and storage need to be clearly delineated. The assay is still a... 

Sometimes, cellular analysis was performed with cells in a flow stream, which is also termed flow cytometry or FACS in the field of cell biology. For instance, human blood cell (WBC, RBC) rheology was studied in channels fabricated on the Si-Pyrex substrates. The channels were either uncoated or coated with albumin [825]. 

Figure 9.3 Mean total lymphocyte count after one course of treatment. Peripheral blood was collected from individuals administered test article (circles) or placebo (squares) on a weekly basis, and subject to flow cytometry analysis to determine mean lymphocyte counts. A standard panel of fluorochrome-conjugated antibodies was used to identify the various lymphocyte sub populations. The solid bar indicates the dosing interval.

JoningsLK, AdmunRA, WangWe, Dodder ME Analysis ofhuman platelet glycoproteins Ilb-lIIa and Glanzmann s thrombasthenia in whole blood by flow cytometry. Blocxl 68 173-179,1986. 

However, the flow cytometers are bulky and expansive, and are available only in large reference laboratories. In addition, the required sample volumes are quite large, usually in the 100 pL range. Many clinical applications require frequent blood tests to monitor patients status and the therapy effectiveness. It is highly desirable to use only small amount of blood samples Ifom patients for each test. Furthermore, it is highly desirable to have affordable and portable flow cytometry instruments for field applications, point-of-care applications and applications in resource-limited locations. To overcome these drawbacks and to meet the increasing needs for versatile cellular analyses, efforts have been made recently to apply microfluidics and lab-on-a-chip technologies to flow cytometric analysis of cells. 

Flow Cytometry Analysis. Flow cytometry analysis of blood and tissues has the advantage of allowing enumeration of lymphocyte subsets (see Chapters... 

Figure 9.2-5 Percent of B and T lymphocytes determined by flow cytometry analysis in pregnant cynomolgus macaque and fetal (cord) blood. Each bar represents the mean SEM of data from 14 pregnant macaques and six fetuses.

Preparation of Blood Cells for Flow Cytometry Analysis... 

Flow cytometry analysis of murine myeloid cells was performed on blood cells of C57BL6 mice however, the protocol that here we describe is appropriate for all mouse strains. Blood can be collected either by cardiac puncture or from the retro-orbital sinus, as here described. 

A wart is a skin lesion caused by infection with human papillomavirus (HPV). Contact immunotherapy is one of the many therapeutic options that has been used to treat warts however, the effectiveness of contact immunotherapy differs from patient to patient, and the cause of this variaHon in clinical response is unclear. To assess cytokine changes in patients after contact immunotherapy with squaric acid dibutylester (SADBE), a total of 21 patients with warts and nine healthy control subjects were enrolled in this study [Ib ]. The frequencies of CD3+ T cells expressing interleukin (IL)-4 IL-10, lL-12, tumour necrosis factor-alpha (TNF-a) and interferon-gamma were measured by flow cytometry analysis of peripheral blood at baseline in both patients and controls, and after SADBE treatment in patients. 

Since the 1980s, silicon-based sensors and microelectromechanical devices (MEMS) have been used ex vivo for tasks such as pressure sensing, blood chemistry analysis, flow cytometry and electrophoresis [6]. The term BioMEMS was subsequently coined to describe the wide array of tools developed using silicon as a platform. Extremely sophisticated systems are now available, such as functionalized microcantilevers for molecule adsorption, recognition and quantification, as... 

Fibach, E. Giloh, H. Rachmilewitz, E. A. Gatt, S. Flow cytofluorometric analysis of the uptake of the fluorescent fatty acid pyrenedodecanoic acid by human peripheral blood cells. Cytometry 1988, 9, 525-528. 

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