Attachment of A172 human glioblastoma cells affects calcium signalling: A comparison of image cytometry, flow cytometry, and spectrofluorometry

J. Szollosi, Burt G.F. Feuerstein, W. C. Hyun, M. K. Das, L. J. Marton

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The intracellular free calcium concentration ([Ca2+](i)) of indo-1 loaded A172 human glioblastoma cells stimulated by platelet-derived growth factor (PDGF) was studied in cell suspensions by flow cytometry and spectrofluorometry and in confluent monolayers by laser image cytometry and spectrofluorometry. With all three techniques, the percentage of responsive cells, peak [Ca2+](i), and the duration of response were directly related, and the delay time was inversely related to PDGF dose. The maximum response occurred at a PDGF concentration of about 20 ng/ml. Basal and peak [Ca2+](i) did not differ significantly from method to method even though different calibration procedures were used. Cells in suspension monitored by both spectrofluorometry and flow cytometry displayed significantly shorter calcium responses than attached cells. This did not appear to be a direct effect of trypsinization. Spectral analysis of indo-1 in cytoplasm, 40% glycerol, and aqueous solutions showed significant differences in the isosbestic point and quantum efficiency. Calibration of [Ca2+](i) with spectrofluorometry is more accurate using the ratio of fluorescence intensities than the fluorescence intensities measured at either 405 or 485 nm.

Original languageEnglish (US)
Pages (from-to)707-716
Number of pages10
JournalCytometry
Volume12
Issue number8
StatePublished - 1991
Externally publishedYes

Fingerprint

Image Cytometry
Calcium Signaling
Fluorescence Spectrometry
Glioblastoma
Flow Cytometry
Platelet-Derived Growth Factor
Calibration
Suspensions
Fluorescence
Calcium
Glycerol
Cytoplasm
Lasers

Keywords

  • Cytosolic free calcium concentration
  • indo-1
  • Monolayer
  • platelet-derived growth factor

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Attachment of A172 human glioblastoma cells affects calcium signalling : A comparison of image cytometry, flow cytometry, and spectrofluorometry. / Szollosi, J.; Feuerstein, Burt G.F.; Hyun, W. C.; Das, M. K.; Marton, L. J.

In: Cytometry, Vol. 12, No. 8, 1991, p. 707-716.

Research output: Contribution to journalArticle

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