Flow cytometry references

Therefore, no detailed discussion on the interaction of any liposomes with any particular cell type should be stated here. We refer to several recent publications for a study of interaction of DOPE CHEMS liposomes and COS-7 and HUVEC (109) and for a study on size-dependent uptake of particles into B16-F10 (72). The combination of flow cytometry and a microscopic method (e.g., spectral bio-imaging) turned out to be highly useful both to study the initial mode of internalization and to follow the intracellular fate of liposomes and other particulate carrier systems. 

Refining the standardized measurement referred to above to provide a calibrated scale is a more difficult task, although calibration beads of various types now available facilitate the process. Beads with manufacturer s reported equivalent fluorescein content have been available for some time (Flow Cytometry Standards Corporation, San Juan, Puerto Rico), and these are useful for calibration of measurements using fluorescein-labeled antibodies, although... 

A good reference on the flow cytometry of DNA fragments is Kim Y, Jett JH, Larson EJ, et al. (1999). Bacterial fingerprinting by flow cytometry Bacterial species discrimination. Cytometry 36 324-332. The proposed technique for sequencing DNA is described in a paper by Jett JH, Keller RA, Martin JC, et al. (1989). High-speed DNA sequencing An approach based upon fluorescence detection of single molecules. J. Biomol. Struct. Dynamics 7 301-309. 

In the Preface, I referred to the way flow cytometry has moved over the past three decades in directions that have surprised even those intimately involved in the field. It is obvious that attempts to predict the future of flow cytometry should be left to informal discussions among friends. The temptation to speculate is, however, an impossible one to resist. This chapter should therefore be read in the spirit in which it was written with a large measure of skepticism (preferably among friends, after a good dinner and a good bottle of wine). 

Although books on flow cytometry abound and articles on flow cytometry can be found throughout a great range of publications, the following is a limited list of references that I have found particularly useful for general information on the theoretical basis of flow analysis and as routes into the literature on particular subjects and techniques. 

Shapiro HM (1995). Practical Flow Cytometry, 3rd edition. Wiley-Liss/John Wiley and Sons, Inc, New York. This wonderful book manages to make learning about flow cytometry more enjoyable than you would have thought possible. There are readable sections on most aspects of flow theory and an excellent compendium of references. A fourth edition is, I believe, in the works. 

Acquisition In flow cytometry, acquisition refers to the process of recording the intensity of the photodetector signals from a particle in the transient memory of a computer. Once acquired, the data from a group of particles can be stored permanently on a storage medium from which it can be subjected to analysis. Acquisition and then analysis (in that order) are the two central steps in the flow cytometric procedure. 

Flow Flow is a term that has come to refer, colloquially, to the general technique of flow cytometry. For example Do you use flow to analyze your cells or Flow has revolutionized the study of immunology or I m off to do flow. ... 

Flower Flower is a term of affection in the Northeast of England— and one that could, with a slight change of pronunciation, be adapted to refer to a person who works in the field of flow cytometry. 

G0/G1 In flow cytometry, the term refers to cells that have the 2C (diploid) amount of DNA and are therefore either not cycling (GO) or have just completed cytokinesis and have not yet begun again to make more DNA in preparation for a new cycle (Gl). 

Probe Probe is a general term used, in flow cytometry, to refer to any chemical that fluoresces when it reacts or complexes with a specific class of molecules and therefore can be used to assay that molecule quantitatively. Propidium iodide and acridine orange are probes for nucleic acid because they complex specifically with nucleic acids and fluoresce brightly when they have... 

Fig. 1. Increasing reference to flow cytometry in the medical literature over the past three decades. The development of flow cytometers antedates the use of the term itself.

Aptamers attached to a fluorescent reporter are useful for quantification of a cell surface antigen on living cells by flow cytometry (see references [21, 38]). 

Various methods exist for the estimation of genome size. The three most common methods are chemical extraction, densitometry, and flow cytometry. Although each of these techniques has advantages and disadvantages, the method that appears to have the greatest potential is flow cytometry. Therefore, flow cytometric analysis of genome size is presented as the method of choice for genome size comparisons. If, however, one is interested in the other methods, several excellent references provide much... 

We do not describe in details the immunological methods (immunization, Gr release assay, ELISPOT, ELISA and flow cytometry) used for the analysis of the immune responses induced by the liposome vaccines (Subheading 3.2). For comprehensive information, we refer to our publications (21-23) and to the related literature. 

The samples were processed and stained with propidium iodide according to the method described in Ref. [14.8]. Flow cytometry DNA analysis was performed using a FACScan cytometer. The fluorescence intensity of the DNA labelled with propidium iodide was processed using CellQuest software, and the histograms of the DNA content were analysed using ModFit software. For each sample, 20 000 events were acquired. The DI was calculated with reference to the DNA diploid from the same sample. 

This table is intended to hold results of assays testing compounds in reg-istry.structure for activity as human immunodeficiency virus (HIV) protease inhibitors. As new assays are added, the test results can be added to newly created tables with similar definitions. For example, there might be tables for HIV reverse transcriptase inhibitors stored in a table named hiv.rt. Other assay results might be stored in new schemas, for example, fpr.htfc for high-throughput flow cytometry results for the formyl peptide receptor (FPR), or f pr.ca for FPR cell adhesion assay results. Each of these tables would have columns of data named and typed appropriately for each assay. Each table would have a column containing a compound id that references compounds in the registry, structure table. 

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. 

Quasi-quantitative assays traditionally include measures of enzymatic or ligandbinding activity, as in flow cytometry and anti-drug antibody assays [9]. One of the common characteristics of these assays is the lack of a true reference standard, where reference standards are poorly characterized, do not completely represent native protein, or differ from native proteins in terms of potency or immunoreactivity. As stated above, if the analytical response is continuous across the range in question, the analytical results can be expressed in terms of a characteristic of known test samples. The following is one example an ELISA qualified as a quasi-quantitative assay because it could not be validated as a relative quantitative assay. 

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