What are the disadvantages of Bright-field microscopy?
Thin objects are in culture almost invisible if they are not stained or observed in phase contrast or in contrast interference (Nomarski). Immuno Histological dyes, although numerous, make it possible to
label components like cellulose, lignin, organelles, organisms,bacteria but sorely lacking in specificity. In situ hybridization offers good specificity a lot of background noise. On the other hand
by dusts, stains of dyes, debris which are not passed to the sample in value.
What are the Advantages of Fluorescence?
Fluorescence offers a wide range of tools that can mark the moment
an element, an organism or a protein. On the other hand, it offers a strong contrast because the object illuminates on a black background, facilitating the detection of interest structures. These molecules are called fluorochromes or fluorophores and are of different kinds like FITC or Alexa 488.
What is the principle of fluorescence?
The Energy E post photon will have a little bit of energy under different forms like radiation, internal conversion before finding
its state of rest by releasing the remaining energy in the form of a photon.
Thus, the released photon is less energetic than the photon incident.
According to Planck’s law, the wavelength (color) is inversely proportional to the energy. The photon emitted from a long incident length.
The confocal Microscope enables to see much smaller than the bright flied microsope. Olympus is the most used supplier of Confocal Microscopes nut Leica has them too.