Why can confocal microscopy not be used for deep tissue imaging?
This is because refractive index discontinuities within the biological tissue still result in scattering of light (Tuchin, 2005b).
Consequently, even when sample and immersion media are index matched, signal levels obtained with confocal microscopy rapidly decrease with depth..
Why is it called confocal microscopy?
In contrast, a confocal microscope uses point illumination (see Point Spread Function) and a pinhole in an optically conjugate plane in front of the detector to eliminate out-of-focus signal – the name “confocal” stems from this configuration.
Why is confocal microscopy better than fluorescence microscopy?
Confocal microscopy offers several distinct advantages over traditional widefield fluorescence microscopy, including the ability to control depth of field, elimination or reduction of background information away from the focal plane (that leads to image degradation), and the capability to collect serial optical …
What is the advantage of confocal microscopy?
Confocal microscopy offers several advantages over conventional optical microscopy, including shallow depth of field, elimination of out-of-focus glare, and the ability to collect serial optical sections from thick specimens.
How does confocal microscopy work?
Similar to the widefield microscope, the confocal microscope uses fluorescence optics. Instead of illuminating the whole sample at once, laser light is focused onto a defined spot at a specific depth within the sample. … By scanning the specimen in a raster pattern, images of one single optical plane are created.
What do laser scanning confocal microscopes observe?
Confocal laser scanning microscopy (CLSM) allows optical slicing through tissue. By eliminating out-of-focus images, CLSM affords greater spatial resolution in living tissue and allows visualization of living structures as small as dendritic spines (Fig.