Abstract
The Microfluidic Optical Stretcher (MOS) has previously been shown to be a versatile tool to measure mechanical properties of single suspended cells. In this study we combine optical stretching and fluorescent calcium imaging. A cell line transfected with a heat sensitive cation channel was used as a model system to show the versatility of the setup. The cells were loaded with the Ca(2+) dye Fluo-4 and imaged with confocal laser scanning microscopy while being stretched. During optical stretching heat is transferred to the cell causing a pronounced Ca(2+) influx through the cation channel. The technique opens new perspectives for investigating the role of Ca(2+) in regulating cell mechanical behavior.
Highlights
Laser based cell analysis is one of the fastest growing areas in medical technologies and has many applications in biology and biophysics
Icard-Arcizet et al combined optical tweezers microrheology with epifluorescence microscopy of GFP-actin and could show that the strengthening of the probed focal adhesions correlated with a recruitment of actin around the attached bead [13]
HEK293 cells, transfected with the heat activated TRPV1 channel and loaded with the fluorescent Ca2+ dye Fluo-4 were optically stretched for 5 s at 700 mW per fiber while the fluorescence signal was recorded with confocal laser scanning microscopy
Summary
Laser based cell analysis is one of the fastest growing areas in medical technologies and has many applications in biology and biophysics. A number of applications and improvements to the setup, such as the integration of the Optical Stretcher into microfluidics, the so called Microfluidic Optical Stretcher (MOS), have been published [10,11,12]. As most optical trapping techniques are performed in commercially available microscopes, combination with fluorescent imaging is straight forward. Icard-Arcizet et al combined optical tweezers microrheology with epifluorescence microscopy of GFP-actin and could show that the strengthening of the probed focal adhesions correlated with a recruitment of actin around the attached bead [13]. In contrast to the tweezers setup, in the Optical Stretcher the optically induced forces act directly on the surface of the suspended cell superseding the attachment of beads to the surface [6, 7]. In this study we combine intracellular Ca2+ imaging with measurements of cellular mechanics
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