Clinical application of flow cytometry: A review

Lærum, Ole Didrik; Farsund, Tore

doi:10.1002/cyto.990020102

Cited by 220 publications

(94 citation statements)

References 102 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The functional properties of cells correlate strongly with these lineage-specific differentiation antigens (14). The subpopulations of B cells (1) and T cells (10) can be quantitated by flow cytometry (FCM) based on the relative expression of two or more cell-surface markers (8), and this approach has been used to define abnormal lymphocyte subsets in a variety of clinical conditions (9,11).…”

mentioning

confidence: 99%

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Lal¹,

Edison²,

Chused³

1988

Cytometry

View full text Add to dashboard Cite

A method to preserve stained human lymphocytes for subsequent cell surface analysis by flow cytometry (FCM) is described. Cells stained with fluorescein isothiocyanate (FITC) and phycoerythrin (€'E)-conjugated monoclonal antibodies and then fixed in 1% paraformaldehyde, followed by extensive washing and resuspension in 1% BSA medium, could be stored at 4°C for at least 2 weeks prior to FCM analysis without significant alteration in the light scatter or fluorescence properties of the cells. Furthermore, the method was also suitable for analyzing lymphocytes that express T-cell activation markers in certain disease conditions. In addition, we have identified monoclonal antibody combinations that discriminate different lymphocyte subsets that are satisfactory for multiparameter analysis after 2 weeks of storage. This method should prove useful for enumerating lymphocyte subsets in field study sites remote from flow cytometry laboratories.Key terms: Paraformaldehyde, filariasis, systemic lupus erythematosusThe lymphocyte population has been shown to be heterogeneous with respect to cell surface antigens (21). The functional properties of cells correlate strongly with these lineage-specific differentiation antigens (14). The subpopulations of B cells (1) and T cells (10) can be quantitated by flow cytometry (FCM) based on the relative expression of two or more cell-surface markers (8), and this approach has been used to define abnormal lymphocyte subsets in a variety of clinical conditions (9, 11).For such clinical studies, fresh lymphocytes (7) have generally had t o be used, since conflicting results have sometimes been observed with cryopreserved cells (31, an observation suggesting that certain differentiation antigens may be cryolabile. Thus, in most studies, cells are stained by conventional immunofluorescence techniques and analyzed either fresh or shortly after paraformaldehyde fixation (13). For studies carried out at sites remote from a flow cytometer, this protocol, however, creates practical problems, since the effect of longterm fixation on FCM analyses has not been well defined.Therefore the purpose of this study was to develop the optimal method of preserving stained cells for as long a time as possible. The basic requirement of such a technique would be that it permit 2-3 weeks storage of the cells prior to analysis without significant alteration of the light scatter and fluorescence properties of the cells. In this report, we describe such a method and define monoclonal antibody reagents satisfactory for studies requiring prolonged storage of FCM samples prior to analysis. MATERIALS AND METHODS Study PopulationThirteen individuals were studied. Eight were healthy North Americans, two were normal volunteers from New Delhi, India, two were patients with systemic lupus erythematosus, and one was an Indian patient with tropical pulmonary eosinophilia.Preparation of Fixative A 1.0% (w/v) solution of paraformaldehyde (Eastman Kodak Co., Rochester, NY) was prepared by dissolving paraformaldehyde in phosphat...

show abstract

mentioning

confidence: 99%

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Lal¹,

Edison²,

Chused³

1988

Cytometry

View full text Add to dashboard Cite

show abstract

“…DNA content can be used to evaluate both the degree of malignancy of tumors and the prognosis "ad vitae" for the patients (2,(4)(5)(6)20,22,24). However, available data for brain tumors are limited and contradictory (l0, 11,18,19,22,24,25~.…”

mentioning

confidence: 99%

Histological grading, DNA content, cell proliferation and survival of patients with astroglial tumors

Zaprianov

Christov²

1988

Cytometry

View full text Add to dashboard Cite

-Light microscopy, image cytometry (ICM), and flow cytometry (FCIM) were used to study the degree of differentiation, DNA content, and S-phase of astrocytomas and glioblastoma multiforme in 102 patients. The postoperative real survival time (RST) was also studied.Using TCM, 62 astrocytomas were investigated. Grade I astrocytomas were composed of DNA-diploid cell lines, while grade I11 and glioblastoma multiforme consisted predominantly of DNA-aneuploid lines. Moderately differentiated astrocytomas were divided as follows: 14 DNA-diploid and 18 DNA-aneuploid. Forty astrocytomas were studied by FCM. Using the DNA index (DI) value, cases with abnormal DNA cell lines were established in all astrocytomas, with their number increasing in grades I1 and I11 astrocytomas.FCM indicated the same subdivision of moderately differentiated astrocytomas: I2 with DNA-diploid and 12 with DNA-aneuploid stem lines. Patients with DNA-diploid cell lines in the astrocytomas and low Sfraction survived longer than patients with abnormal DNA cell populations and higher S-fraction.The results from this study indicate that, together with the degree of differentiation of astroglial tumors, the appearance of cell lines with abnormal DNA value and higher S-fractions also have prognostic value.

show abstract

“…Nevertheless, a relationship between nuclear DNA content and clinical course has been reported for several carcinoma entities. Thus, diploid tumors have been shown to be associated with a better prognosis than hyperploid (Barlogie et al 1983;Laerum and Farsund 1981;Tribukait 1987). Since this does not apply to all malignant entities, the relationship between DNA content and clinical course has to be investigated for each tumor entity separately.…”

Section: Dna Cytophotometrymentioning

confidence: 99%

DNA cytometry of osteosarcoma

Bauer

1988

Acta Orthopaedica Scandinavica

View full text Add to dashboard Cite

Clinical application of flow cytometry: A review

Cited by 220 publications

References 102 publications

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Histological grading, DNA content, cell proliferation and survival of patients with astroglial tumors

DNA cytometry of osteosarcoma

Contact Info

Product

Resources

About