Differential Light Scattering from Spherical Mammalian Cells

Brunsting, A.; Mullaney, P. F.

doi:10.1016/s0006-3495(74)85925-4

Cited by 240 publications

(141 citation statements)

References 24 publications

(19 reference statements)

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“…Figure 2B shows that at the same time, however, the median FLS intensity of the samples decreased by a factor of 5 . We consider this large decrease of the forward light scatter, resulting from intracellular gas vesicles, anomalous, in view of the fact that forward light scatter of small biological particles is normally associated mainly with cell size, and to a-much lesser extent with internal structure (2,11,12,17,22,24). Electrical volume measurements and microscopic observations showed, however, that the diameters of the cells did not change.…”

Section: Flow Cytometrymentioning

confidence: 86%

Anomalous behaviour of forward and perpendicular light scattering of a cyanobacterium owing to intracellular gas vacuoles

1987

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Extinction, absorption, and forward and perpendicular light scatter of the blue-green alga Microcystis aeruginosa with different amounts of intracellular gas vacuoles were determined. The amount of gas vacuoles in the cells was controlled by application of pressure. The presence of the gas vacuoles caused a tenfold increase in perpendicular light scatter, and a fivefold decrease in forward light scatter as measured by flow cytometry. Chlorophyll fluorescence showed a 16% decrease. The presence of gas vacuoles did not affect the size of the algae. The absorption spectrum of Microcystis aeruginosa was slightly raised but practically not distorted by the gas vacuoles. The attenuation spectrum, a measure for light extinction by the algal cells, was significantly distorted. The increase of perpendicular light scatter intensity of the cells is a direct consequence of the relatively high scatter of each vacuole, whereas the forward light scatter decrease is attributed to a lower phase-shift factor p of the whole cells, caused by the intact gas vacuoles.

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Section: Flow Cytometrymentioning

confidence: 86%

Anomalous behaviour of forward and perpendicular light scattering of a cyanobacterium owing to intracellular gas vacuoles

1987

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“…FAS would be indicative of cell size and RAS of cytoplasmic heterogeneity (1,2,13). For these reasons, an attempt was made to correlate the cytotoxic activity of the macrophages sorted both with their size and with content in acid phosphatase, an enzyme present in cytoplasmic lysosomal granules.…”

Section: Discussionmentioning

confidence: 99%

Characterization and sorting of mouse cytotoxic macrophages by their light scattering properties

et al. 1983

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The light scattering properties of mouse activated macrophages were analyzed by flow cytometry. Peritoneal adherent cells from B. abortus treated animals were found to segregate into two subpopulations as a function of their foward angle and 90" angle light scatter. The cell subpopulations were separated by automatic sorting. The strongly scattering ones contained an elevated proportion of large volume and acid phosphatase rich cells. Their nonspecific cytotoxic activity against tumor cells was more important than that of weakly light scattering cells. Thus, flow cytometry might be helpful to characterize and isolate cytotoxic macrophage populations.Key terms: Flow cytometry, light scattering, cell sorting, activated macrophages, cytotoxicityThe morphological and functional heterogeneity of macrophages has been evidenced in a number of experimental systems. Cell flow analysis also confirmed this notion, showing that several populations of macrophages could be distinguished as a function of their content in RNA (17,18) and in various enzymes (5) or according to their affinity for certain antimacrophage monoclonal antibodies (15).Using cell sorting techniques, flow cytometry also offers the possibility to study biological properties of isolated cell subpopulations. In the present work, mouse activated macrophages were analyzed and sorted as a function of their light scattering properties. Strongly and weakly light scattering cells were separated and their cytotoxic activity, cell volume, and acid phosphatase content were measured. MATERIALS AND METHODS Preparation of Peritoneal Exudate CellsCells were obtained by washing the peritoneal cavity of (C57BL6 x DBAB) F1 hybrid mice injected IP 3 days before with 500 pg of heat-inactivated B. abortus organisms as described previously (10,ll). The cells were left to adhere on microexudate-coated culture flasks as described by Mantovani et al (9). After 20 min of incubation the nonadherent cell fraction was separated. Adherent cells were detached by a 10-min exposure to ethylene-diamine-tetracetate (EDTA). Their viability was 92% as demonstrated by a Trypan blue exclusion test. Cells were suspended at 40 x 106/ml in RPMI 1640 medium (Difco) before cell flow measurements and sorting. Flow CytometryThe cytofluorograf system 50 H (Ortho Diagnostic Instruments; Westwood, MA) was uscd. The apparatus was standardized daily for scatter using a suspension of glutaraldehyde-fixed calf thymocytes. The suspensions of fresh peritoneal cells, maintained at 4"C, were previously filtered through 50-pm nylon filter. An argon laser beam of wavelength 488 nm and output energy 500 mW was used to illuminate the cells. Light scattering at 90" and at low forward angle (2"-20") was measured. Right angle scatter (RAS) and forward angle scatter (FAS) signals were displayed as a cytogram on a multichannel distribution analyzer (Model 2103, Digital Data Processing) that could also display frequency distribution histograms of cell number against one of the parameters FAS or RAS. Ten or twenty thousan...

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“…Clearly, we observe evident intracellular contrast resting on the refractive index distribution of hMSCs in our system. For example, the nucleus, a membrane-bound organelle that is densely comprised of nucleic acids and proteins associated with a substantially higher refractive index than the surrounding cytoplasm, [ 32 ] can be thus easily visible as presented in red corresponding to the greatest shift of resonance frequencies and the strongest refl ection intensity. Besides, the other colored parts mainly refer to the cytoplasm, corresponding to the smaller shift of resonant frequencies stemming from the lower refractive index of the cytoplasm.…”

Section: Methodsmentioning

confidence: 99%

Label‐Free, Coupler‐Free, Scalable and Intracellular Bio‐imaging by Multimode Plasmonic Resonances in Split‐Ring Resonators

et al. 2012

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Differential Light Scattering from Spherical Mammalian Cells

Cited by 240 publications

References 24 publications

Anomalous behaviour of forward and perpendicular light scattering of a cyanobacterium owing to intracellular gas vacuoles

Anomalous behaviour of forward and perpendicular light scattering of a cyanobacterium owing to intracellular gas vacuoles

Characterization and sorting of mouse cytotoxic macrophages by their light scattering properties

Label‐Free, Coupler‐Free, Scalable and Intracellular Bio‐imaging by Multimode Plasmonic Resonances in Split‐Ring Resonators

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