What are the advantages and disadvantages of a confocal microscope?

Advantages of confocal microscopy include rapid, noninvasive technique allowing early diagnosis and management and high resolution images[2] as compared to CT scan, MRI and USG for dermatological use. Disadvantages of confocal microscopy include its high cost and relatively smaller field of vision.

What is the advantage to using a confocal microscope?

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.

What is special about confocal microscopy?

Confocal microscopy provides the capacity for direct, noninvasive, serial optical sectioning of intact, thick, living specimens with a minimum of sample preparation as well as a marginal improvement in lateral resolution compared to wide-field microscopy.

Is confocal microscopy difficult?

A confocal microscope gives us a clear examination and 3D reconstruction of thick specimens due to effective Z-axis scanning. However, it can be time-consuming to use a confocal microscope (depending on the scanning speed) and it has a more complicated image acquisition procedure compared to a widefield microscope.

What are the disadvantages of confocal microscopy?

Disadvantages of confocal microscopy are limited primarily to the limited number of excitation wavelengths available with common lasers (referred to as laser lines), which occur over very narrow bands and are expensive to produce in the ultraviolet region.

What is the resolution of confocal microscopy?

When optimally used, confocal microscopes may reach resolutions of 180 nm laterally and 500 nm axially, however, axial resolution in depth is often impaired by spherical aberration that may occur due to refractive index mismatches.

Which is the greatest advantage of using confocal microscopy?

Advantages and Disadvantages of Confocal Microscopy The primary advantage of laser scanning confocal microscopy is the ability to serially produce thin (0.5 to 1.5 micrometer) optical sections through fluorescent specimens that have a thickness ranging up to 50 micrometers or more.

What is the difference between confocal and fluorescence microscopy?

The fluorescence microscope allows to detect the presence and localization of fluorescent molecules in the sample. The confocal microscope is a specific fluorescent microscope that allows obtaining 3D images of the sample with good resolution. This allows to reconstruct a 3D image of the sample.

What is resolution limit?

The limit of resolution (or resolving power) is a measure of the ability of the objective lens to separate in the image adjacent details that are present in the object. It is the distance between two points in the object that are just resolved in the image. Thus an optical system cannot form a perfect image of a point.

What is the resolution of fluorescence microscopy?

Spatio-temporal visualization of cellular structures by fluorescence microscopy has become indispensable in biology. However, the resolution of conventional fluorescence microscopy is limited by diffraction to about 180 nm in the focal plane and to about 500 nm along the optic axis.

Which is an advantage of confocal microscopy over widefield microscopy?

Confocal microscopy offers several advantages over conventional widefield optical 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 sections from thick specimens.

What are the limitations of point scanning confocal microscopy?

A key limitation in the use of point-scanning confocal microscopy for live cell imaging applications is the speed of image acquisition, which may be too slow to obtain information about rapid biological processes. Spinning disk systems provide an alternative means of obtaining a full-frame high-speed confocal image in real-time.

How many optical sections can be used in confocal microscopy?

Confocal microscopy allows 1–2 μm optical sectioning of 50–100 μm thick sections with rejection of out-of-focus light. The Z-stack of optical sections can then be used to obtain high-contrast three-dimensional reconstructions for examination of the interrelationships between stained structures in which any fine detail appears perfectly “in focus.”

How is confocal microscopy used in biological imaging?

Confocal microscopy is an optical imaging technique commonly employed to increase the optical resolution and contrast of an image. A routine problem encountered in imaging three-dimensional biological samples with compound light or fluorescence microscopes is that light is captured in multiple focal planes to create a single two-dimensional image.