Confocal restricted-height imaging of suspension cells (CRISC) in a PDMS microdevice during apoptosis.
We have monitored and imaged cell death induced in human leukemic U937 cells over time using three-color confocal imaging. Three different apoptotic inducers, anti-Fas, TNF-alpha and Etoposide were used. Individual cascaded events such as loss of mitochondrial transmembrane potential, exposure of phosphatidyl-serine, membrane blebbing and permeabilization of the cell membrane have been observed in real time with different individual cells. From the results, an interesting heterogeneicity in the apoptotic phenotype has been observed. The CRISC method is easy to use and provides biologist with a powerful additional tool to study in real-time processes of several hours of duration such as apoptosis. We predict that the period of cell viability obtained after protein coating of the PDMS devices (>80 h) will also allow monitoring of other biological processes of longer duration or long onset time, such as mitosis, phagocytosis and differentiation.
Muñoz-Pinedo
Cristina
C
La Jolla Institute for Allergy and Immunology, San Diego, CA, USA. cmunoz@liai.org
Green
Douglas R
DR
van den Berg
Albert
A
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D058846:Antibodies, Monoclonal, Murine-Derived; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004867:Equipment Design; D005047:Etoposide / Q000494:pharmacology; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D010316:Particle Size; D012680:Sensitivity and Specificity; D013499:Surface Properties; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D020298:U937 Cells
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D004129:Dimethylpolysiloxanes; D014409:Tumor Necrosis Factor-alpha; C411023:anti-Fas monoclonal antibody; D005047:Etoposide