Fluorescence microscope
A fluorescence microscope is an optical microscope that uses fluorescence and phosphorescence to study organic or inorganic substances.[1][2] "Fluorescence microscope" means any microscope that uses fluorescence to make a picture. This applies whether it is a more simple set up or a more complicated design.[3]
Most fluorescence microscopes, especially those used in the life sciences, are of the epifluorescence design shown in the diagram. Light of the excitation wavelength illuminates the specimen through the objective lens. The fluorescence emitted by the specimen is focused onto the detector. The dichroic beamsplitter acts as a wavelength specific filter, transmitting fluoresced light through to the eyepiece or detector, but reflecting any remaining excitation light back towards the source.
Fluorescence Microscope Media
An upright fluorescence microscope (Olympus BX61) with the fluorescence filter cube turret above the objective lenses, coupled with a digital camera.
Fluorescence and confocal microscopes operating principle
3D-animation of the diatom Corethron sp.Displays overlays from four fluorescent channels (a) Green: [DiOC6(3) fluorescence] - stains cellular membranes indicating the core cell bodies(b) Cyan: [PLL-A546 fluorescence] - generic counterstain for visualising eukaryotic cell surfaces(c) Blue: [Hoechst fluorescence] - stains DNA, identifies nuclei(d) Red: [chlorophyll autofluorescence] - resolves chloroplastsThe animation starts by overlaying all available fluorescent channels, and then clarifies the visualisation by switching channels on and off 
A z-projection of an osteosarcoma cell, stained with phalloidin to visualise actin filaments. The image was taken on a confocal microscope, and the subsequent deconvolution was done using an experimentally derived point spread function.
Super-resolution microscopy: Single YFP molecule detection in a human cancer cell. Typical distance measurements in the 15 nm range measured with a Vertico-SMI/SPDMphymod microscope
Super-resolution microscopy: Co-localization microscopy (2CLM) with GFP and RFP fusion proteins (nucleus of a bone cancer cell) 120.000 localized molecules in a wide-field area (470 µm2) measured with a Vertico-SMI/SPDMphymod microscope
Fluorescence microscopy of DNA Expression in the Human Wild-Type and P239S Mutant Palladin.
References
- ↑ Spring K.R. & Davidson M.W. "Introduction to Fluorescence Microscopy". Nikon MicroscopyU. Retrieved 2008-09-28.
- ↑ "The Fluorescence Microscope". Microscopes help scientists explore hidden worlds. The Nobel Foundation. Retrieved 2008-09-28.
- ↑ Juan Carlos Stockert & Alfonso Blázquez-Castro (2017). Fluorescence microscopy in life sciences. Bentham Science Publishers. ISBN 978-1-68108-519-7. Archived from the original on 14 May 2019. Retrieved 17 December 2017.