E. Electronic image processing B 5 um Figure 3-12. Extending the limits of detection. Light-microscope images of unstained microtubules that have been visualized by differential-interference-contrast microscopy followed by electronic image processing.(A) The original unprocessed image. (B) The final result of an electronic process that greatly enhances contrast and reduces"noise. (Courtesy of Bruce Schnapp
Figure 3-12. Extending the limits of detection. Light-microscope images of unstained microtubules that have been visualized by differential-interference-contrast microscopy followed by electronic image processing. (A) The original unprocessed image. (B) The final result of an electronic process that greatly enhances contrast and reduces "noise." (Courtesy of Bruce Schnapp.) E. Electronic image processing
video-enhance(contrast) microscopy (b) Figure A-17 Computer-Enhanced Digital Video Microscopy image resulting from electronic contrast enhancement of the original This series of micrographs shows how computers can be used to enhance image (which appeared to be empty l. (b) The background of the enhanced images obtained with light microscopy. In this example, an image of image in (a), which is then (c)subtracted from image (a leaving only the several microtubules, which are too small to be seen with unenhanced microtubules. (d) The final, detailed image resulting from electronic light microscopy are processed to make them visible in detail. la)The averaging of the separate images processed as shown in a-c observing living specimens; greatly increase the contrast of an image so that very small objects become visible
Video-enhance(contrast) microscopy ❖Observing living specimens; ❖Greatly increase the contrast of an image so that very small objects become visible
F The confocal confocal tholes microscope dichroic GFP can be used obiective to study dynamic point of focus processes as they fluorescent specimen is illuminated emitted fluorescent emitted light from out occur in a living with a focused point of light from ht from in-focus of-focus point is out of 0 pinhole point is focused at focus at pinhole and cell hole and reaches is largely excluded detector from detector Figure 3-13. The confocal microscope. This diagram shows that the basic arrangemen of optical components is similar to that of the standard fluorescence microscope except that a laser is used to illuminate a small pinhole whose image is focused at a single point in the specimen(A). Fluorescence from this focal point in the specimen is focused at a second pinhole (b). light from elsewhere in the specimen is not focused here and therefore does not contribute to the final image(C). By scanning the beam of light across the specimen, a very sharp two-dimensional image of the exact plane of focus is built up that is not significantly degraded by light from other regions of the specimen
Figure 3-13. The confocal microscope. This diagram shows that the basic arrangement of optical components is similar to that of the standard fluorescence microscope except that a laser is used to illuminate a small pinhole whose image is focused at a single point in the specimen (A). Fluorescence from this focal point in the specimen is focused at a second pinhole (B). Light from elsewhere in the specimen is not focused here and therefore does not contribute to the final image (C). By scanning the beam of light across the specimen, a very sharp two-dimensional image of the exact plane of focus is built up that is not significantly degraded by light from other regions of the specimen. F. The confocal microscope 32.mov GFP can be used to study dynamic processes as they occur in a living cell
(A) B) Figure 3-14. Comparison of conventional and confocal fluorescence microscopy. These two micrographs are of the same intact gastrula-stage Drosophila embryo that has been stained with a fluorescent probe for actin filaments. The conventional, unprocessed image(a)is blurred by the presence of fluorescent structures above and below the plane of focus. In the confocal image(B), this out-of- focus information is removed, which results in a crisp optical section of the cell in the embryo
Figure 3-14. Comparison of conventional and confocal fluorescence microscopy. These two micrographs are of the same intact gastrula-stage Drosophila embryo that has been stained with a fluorescent probe for actin filaments. The conventional, unprocessed image (A) is blurred by the presence of fluorescent structures above and below the plane of focus. In the confocal image (B), this out-offocus information is removed, which results in a crisp optical section of the cell in the embryo
2. Electron microscope Figure 3-16. Limit of resolution of the electron microscope. electron micrograph of a thin layer of gold showing the individual files of atoms in the crystal as bright spots. The distance between adjacent files of gold atoms is about 0.2 nm(2 A)
Figure 3-16. Limit of resolution of the electron microscope. Electron micrograph of a thin layer of gold showing the individual files of atoms in the crystal as bright spots. The distance between adjacent files of gold atoms is about 0.2 nm (2 Å). 2. Electron microscope