Process
STED functions by depleting specific regions of the sample while leaving a center focal spot active to emit fluorescence. This focal area can be engineered by altering the properties of the pupil plane of the objective lens. The most common early example of this diffractive optical elements, or DOEs, is a doughnut shape used in two dimensional lateral confinement shown below. The red zone is depleted, while the green spot is left active. This DOE is generated by a circular polarization of the depletion laser, combined with a helical phase ramp. The lateral resolution of this DOE is typically between 30 and 80 nm. However, values down to 2.4 nm have been reported. Using different DOEs, axial resolution on the order of 100 nm have been demonstrated. A modified Abbe’s equation describes this sub diffraction resolution as: D=λ/(2nsinα*sqrt(1+I/Isat)). To optimize the effectiveness of STED, the destructive interference in the center of the focal spot needs to be as close to perfect as possible. That imposes certain constraints on the optics that can be used.
Read more about this topic: STED Microscopy
Famous quotes containing the word process:
“Interior design is a travesty of the architectural process and a frightening condemnation of the credulity, helplessness and gullibility of the most formidable consumers—the rich.”
—Stephen Bayley (b. 1951)
“Science and art are only too often a superior kind of dope, possessing this advantage over booze and morphia: that they can be indulged in with a good conscience and with the conviction that, in the process of indulging, one is leading the “higher life.””
—Aldous Huxley (1894–1963)
“Thinking is seeing.... Every human science is based on deduction, which is a slow process of seeing by which we work up from the effect to the cause; or, in a wider sense, all poetry like every work of art proceeds from a swift vision of things.”
—Honoré De Balzac (1799–1850)