[49] Confocal Microscopy Flashcards
What is confocal microscopy?
A technique in optical microscopy where a laser light source is used to obtain high-resolution images and reconstructions of three-dimensional specimens.
How does confocal microscopy work?
It uses a point illumination and a pinhole in an optically conjugate plane in front of the detector to eliminate out-of-focus signal - mainly on the edge of the specimen.
What is a laser in confocal microscopy?
A coherent light source used for illumination.
What is the advantage of confocal microscopy over conventional fluorescence microscopy?
It provides depth selectivity and eliminates out-of-focus glare.
What is the key feature of confocal microscopy that provides depth selectivity?
The pinhole at the detector.
Why is fluorescence used in confocal microscopy?
It allows specific labeling of components in the cell.
What is a fluorophore?
A fluorescent chemical compound that can re-emit light upon light excitation.
What is meant by “optical sectioning” in confocal microscopy?
The ability to focus on a particular depth within the specimen and capture images one layer at a time.
What is z-stacking in confocal microscopy?
The process of taking multiple 2D images at different depths to create a 3D image stack.
Why is the laser scanning confocal microscope (LSCM) widely used?
Because it allows for control of depth of field, less background light, and the ability to collect serial optical sections from thick specimens.
LSCM - Initials
Laser Scanning Confocal Microscope
What is the function of the objective lens in confocal microscopy?
To collect the fluorescent light and focus it through the pinhole.
What is photobleaching and how is it a disadvantage in confocal microscopy?
The photochemical destruction of a fluorophore, a problem in confocal microscopy because of the intense light illumination.
What is phototoxicity in the context of confocal microscopy?
The damage caused to live cells due to the absorption of light, leading to harmful chemical reactions.
What kind of samples can be observed with confocal microscopy?
Both fixed and living cells can be observed.
What is immuno-labeling in the context of confocal microscopy?
A technique where antibodies tagged with fluorescent dyes are used to label specific components of the cell.
What is “resolution” in the context of confocal microscopy?
The smallest detail of an object that can be observed and recorded by the microscope.
How can confocal microscopy be used in multi-color imaging?
By using different fluorophores that emit at different wavelengths, multiple biological structures can be imaged simultaneously.
What is the use of “DIC” in confocal microscopy?
Differential Interference Contrast (DIC) can be used to generate a pseudo three-dimensional image of a sample, often used in combination with confocal images to give context to the fluorescent data.
DIC - Initials
Differential Interference Contrast
