An inverted fluorescence microscope is an inverted compound light microscope equipped with modules to internally separate the different wavelengths of light, and direct them to respective detection channels. Its main components include a light source, an objective, a filter cube and a detection unit, which can be an eyepiece or a camera (Fig.1). Filter cube is a component which separates a fluorescence microscope from a light microscope. It consists of one excitation filter, one emission filter and a dichroic or dichromatic mirror (Fig. 2)
Dichromatic mirror forms the heart of fluorescence microscopy, because it allows some wavelengths of light to pass through it, while reflecting some other wavelengths. This becomes essential for separation …show more content…
7), which functions both as a condenser and an objective. The light beam from the source is expanded to the size of the rear aperture of the objective and then passed though the exciation filter to make it monochromatic. Using the dichroic mirror, the light is diverted into the objective This forms an intense diffraction-limited spot inside the sample, where the sample absorbs the exitation wavelength of light and emits the red shifted emission wavelength (the phenomenon of fluorescence). Since the diffraction limited sopt is formed on the focal plane of the objective, the emmitted light is gathered back by the objective and is passed on towards the dichroic mirror, which allows it to transmit. Light then passes through the pinhole and is gathered by the detector, usually a CCD camera or a …show more content…
11), are crucial in forming irreversible aggregates. They do not segregate upon changing the environment such as pH or dilution. This type of cluster is different from the stoichiometric interconnected group of folded proteins[28] and protein crystallization [27,13]
In general, monomeric form of protein is the starting reactant of protein aggregation process. In the occurrence of cases of unwanted aggregation, there is a matter in question that which type of monomer’s form (native, active, denatured) takes initiative on the path of aggregation. There are numerous examples which confirm that the misfolded proteins or partially folded intermediates are the first steps in the mechanism of unwanted protein aggregation.[33-39]
As far as kinetics of protein aggregation is concerned, it is believed that the protein aggregates trap the misfolded proteins to further increase the size of aggregates. For this reason the product formation proceeds at an exponential rate, till all the substrates (misfolded protein species) are consumed. This gives an explanation why protein aggregation problem is aggravated at later stages and why is it important to detect the aggregation precursors at an early stage for prevention of further
Dichromatic mirror forms the heart of fluorescence microscopy, because it allows some wavelengths of light to pass through it, while reflecting some other wavelengths. This becomes essential for separation …show more content…
7), which functions both as a condenser and an objective. The light beam from the source is expanded to the size of the rear aperture of the objective and then passed though the exciation filter to make it monochromatic. Using the dichroic mirror, the light is diverted into the objective This forms an intense diffraction-limited spot inside the sample, where the sample absorbs the exitation wavelength of light and emits the red shifted emission wavelength (the phenomenon of fluorescence). Since the diffraction limited sopt is formed on the focal plane of the objective, the emmitted light is gathered back by the objective and is passed on towards the dichroic mirror, which allows it to transmit. Light then passes through the pinhole and is gathered by the detector, usually a CCD camera or a …show more content…
11), are crucial in forming irreversible aggregates. They do not segregate upon changing the environment such as pH or dilution. This type of cluster is different from the stoichiometric interconnected group of folded proteins[28] and protein crystallization [27,13]
In general, monomeric form of protein is the starting reactant of protein aggregation process. In the occurrence of cases of unwanted aggregation, there is a matter in question that which type of monomer’s form (native, active, denatured) takes initiative on the path of aggregation. There are numerous examples which confirm that the misfolded proteins or partially folded intermediates are the first steps in the mechanism of unwanted protein aggregation.[33-39]
As far as kinetics of protein aggregation is concerned, it is believed that the protein aggregates trap the misfolded proteins to further increase the size of aggregates. For this reason the product formation proceeds at an exponential rate, till all the substrates (misfolded protein species) are consumed. This gives an explanation why protein aggregation problem is aggravated at later stages and why is it important to detect the aggregation precursors at an early stage for prevention of further