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ABSTRACT The development of fluorescent labels and powerful Imaging technologies In the last two decades has revolutionized the field of fluorescence microscopy, which Is now widely used In diverse scientific fields from biology to biomedical and materials science. Fluorescence microscopy has also become a standard technique In research laboratories working on Drosophila melanogaster as a model organism. Here, we review the principles of fluorescence microscopy technologies from wide-field to Super-resolution microscopy and Its application In the Drosophila research field.
KEYWORDS imaging; labeling; transgenic reporters; FlyBook
Milestones in the History of Fluorescence Microscopy
FOR almost 300 years, since the invention of the light microscope in the 17th century, microscopists have been limited to studies of unstained cells using transmitted white light. The phenomenon that organic and inorganic substances, such as chlorophyll and vitamins, are capable ofabsorbing light and emitting photons at a higher wavelength, named "fluorescence" by Stokes in 1852, has brought about the era of fluorescence light microscopy. The first fluorescence microscopes were built by August Köhler (in 1904), Carl Reichert and Oskar Heimstädt (in 1911), and Carl Zeiss and Heinrich Lehmann (in 1913), and fluorescence microscopy serves researchers in all fields of life sciences to this day.
In the early days of fluorescence microscopy, researchers were limited to specimens that autofluoresce. In the 1930s, fluorescent stains to label nonfluorescing tissues were first introduced by Max Haitinger and, in the 1950s, Albert Coons and Nathan Kaplan developed a method using antibodies that are coupled to fluorescent dyes to detect antigens in tissues. The greatest leap forward in fluorescence microscopy has been the discovery of the Green Fluorescent Protein (GFP) in the jellyfish Aequorea victoria by Osamu Shimomura in 1962 (Shimomura et al. 1962). The sequencing and cloning of GFP in 1992 (Prasher et al 1992), together with the generation of the first transgenic organism by Martin Chalfie 2 years later (Chalfie et al. 1994), enabled endogenous protein labeling in living organisms that revolutionized biological research. The engineering of GFP variants by Roger Tsien in 1994 (Heim et al. 1994) allowed simultaneous visualization of multiple cellular components. In 2008, these milestones in fluorescence microscopy were honored by the Nobel Committee, which awarded Osamu Shimomura, Martin Chalfie, and Roger Y. Tsien the Nobel Prize in Chemistry...