Confocal microscopy is an optical microscopy technique providing benefits across an expansive set of research areas. Benefits of confocal microscopy include:
A higher control of depth of field
Reduction in out of focus blur outside of the focal plane of interest
An improved ability to penetrate thick biology (e.g., 3D cell culture, whole organism, tissue sections, etc.)
Both qualitative and quantitative benefits are clear to see in the U87 spheroid sample shown above. The confocal image is clearer, with less out of focus haze and demonstrates better quantification of cells across the entire spheroid.
Spinning Disk Confocal Microscopy:
Since the inception of confocal microscopy, a range of technologies have been used to achieve confocality within a microscopic image of a specimen. These techniques typically make use of a pinhole within the optical path. The BioTek Cytation C10 uses a rotating spinning disk with an array of pinholes. These spinning disk confocal systems provide distinct advantages over single point scanning systems allowing users to use decreased excitation energy to illuminate their sample and reduce phototoxicity and photobleaching. This is important when imaging live samples where sample perturbation with illumination light can be phototoxic.
Cytation C10 confocal optical path with two (60 µm and 40 µm) spinning disk, confocal filter cubes and Hamamatsu scientific CMOS camera. Two spinning disk pinhole sizes offer users confocality over a wide range of magnifications.
Cytation C10 examples:
The benefits of confocal microscopy for many sample types can be seen in the examples below.
3D renderings of an adult mouse retina tissue section z-stack showing interaction of astrocytes with retinal vasculature
4 µm beads imaged in widefield and confocal show superior axial resolution in confocal because out of focus light is eliminated by the pinhole.
Movie shows embryonic zebrafish neuronal axons (red) and axial muscle cells (green), rotating in the y-axis, as seen in the Gen5 3D viewer. Improved resolution is clearly visible in the confocal images.
A z-stack and 3D rendering of an intestinal organoid model imaged with the spinning disk confocal optics of a Cytation C10. Caco-2 cells seeded and allowed to form a tube in the Mimetas OrganoPlate.
Confocal Technology is available in:
Agilent BioTek Cytation C10 confocal imaging reader