Problem 1P Scanning Confocal Microscopy Although modern microscopes are marvels of optical engineering, their basic design is not too different from the 1665 compound microscope of Robert Hooke. Recently, advances in optics, lasers, and computer technology have made practical a new kind of optical microscope, the scanning confocal microscope. This microscope is capable of taking images of breathtaking clarity. The figure shows the microscope’s basic principle of operation. The left part of the figure shows how the translucent specimen is illuminated by light from a laser. The laser beam is converted to a diverging bundle of rays by suitable optics, reflected off a mirror, then directed through a microscope objective lens to a focus within the sample. The microscope objective focuses the laser beam to a very small (?0.5 ?m) spot. Note that light from the laser passes through other regions of the specimen but, because the rays are not focused in those regions, they are not as intensely illuminated as is the point at the focus. This is the first important aspect of the design: Very intensely illuminate one very small volume of the sample while leaving other regions only weakly illuminated. As shown in the right half of the figure, light is reflected from all illuminated points in the sample and passes back through the objective lens. The mirror that had reflected the laser light downward is actually a partially transparent mirror that reflects 50% of the light and transmits 50%. Thus half of the light reflected upward from the sample passes through the mirror and is focused on a screen containing a small hole. Because of the hole, only light rays that emanate from the brightly illuminated volume in the sample can completely pass through the hole and reach the light detector behind it. Rays from other points in the sample either miss the hole completely or are out of focus when they reach the screen, so that only a small fraction of them pass through the hole. This second key design aspect limits the detected light to only those rays that are emitted from the point in the sample at which the laser light was originally focused. So we see that (a) the point in the sample that is at the focus of the objective is much more intensely illuminated than any other point, so it reflects more rays than any other point, and (b) the hole serves to further limit the detected rays to only those that emanate from the focus. Taken together, these design aspects ensure the detected light comes from a very small, very well-defined volume in the sample. The microscope as shown would only be useful for examining one small point in the sample. To make an actual image, the objective is scanned across the sample while the intensity is recorded by a computer. This procedure builds up an image of the sample one scan line at a time. The final result is a picture of the sample in the very narrow plane in which the laser beam is focused. Different planes within the sample can be imaged by moving the objective up or down before scanning. It is actually possible to make three dimensional images of a specimen in this way. The improvement in contrast and resolution over conventional microscopy can be striking. The images show a section of a mouse kidney taken using conventional and confocal microscopy. Because light reflected from all parts of the specimen reaches the camera in a conventional microscope, that image appears blurred and has low contrast. The confocal microscope image represents a single plane or slice of the sample, and many details become apparent that are invisible in the conventional image. The following questions are related to the passage “Scanning Confocal Microscopy”. A laser beam consists of parallel rays of light. To convert this light to the diverging rays required for a scanning confocal microscope requires A. A converging lens. B. A diverging lens. C. Either a converging or a diverging lens.
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Textbook Solutions for College Physics
Question
Problem 9P
The Fire in the Eye
You have certainly seen the reflected light from the eyes of a cat or a dog at night. This “eye shine” is the reflection of light from a layer at the back of the eye called the tapetum lucidum (Latin for “bright carpet”). The tapetum is a common structure in the eyes of animals that must see in low light. Light that passes through the retina is reflected by the tapetum back through the cells of the retina, giving them a second chance to detect the light.
Sharks and related fish have a very well-developed tapetum. Figure V.2 a shows a camera flash reflected from a shark’s eye back toward the camera. This reflected light is much brighter than the diffuse reflection from the body of the shark. How is this bright reflection created?
Figure V.2 b shows a typical tapetum structure for a fish. (The tapetum in land animals such as cats, dogs, and deer uses similar principles but has a different structure.) The reflection comes from the interfaces between two layers of nearly transparent cells (whose index of refraction is essentially that of water) and a stack of guanine crystals sandwiched between. Light is reflected from the interface at both sides of the stack of crystals. For certain wavelengths, constructive interference leads to an especially strong reflection.
Bright light from a distant source is focused by the lens of a shark’s eye to a point on the retina, as shown in Figure V.2 c. The tapetum reflects these rays back through the lens, where refraction bends them into parallel rays traveling back toward the source of the light. Because the reflected light from the tapetum is directional, it is much brighter than the diffuse reflection from the shark’s body. But the bright reflection is seen by an observer—or a camera—only at or near the source of the flash that produced the reflection.
Light of wavelength 600 nm in air passes into the layer of guanine crystals. What is the wavelength of the light in this layer?
A. 1100 nm
B. 600 nm
C. 450 nm
D. 330 nm
Solution
The first step in solving V problem number trying to solve the problem we have to refer to the textbook question: Problem 9PThe Fire in the EyeYou have certainly seen the reflected light from the eyes of a cat or a dog at night. This “eye shine” is the reflection of light from a layer at the back of the eye called the tapetum lucidum (Latin for “bright carpet”). The tapetum is a common structure in the eyes of animals that must see in low light. Light that passes through the retina is reflected by the tapetum back through the cells of the retina, giving them a second chance to detect the light.Sharks and related fish have a very well-developed tapetum. Figure V.2 a shows a camera flash reflected from a shark’s eye back toward the camera. This reflected light is much brighter than the diffuse reflection from the body of the shark. How is this bright reflection created?Figure V.2 b shows a typical tapetum structure for a fish. (The tapetum in land animals such as cats, dogs, and deer uses similar principles but has a different structure.) The reflection comes from the interfaces between two layers of nearly transparent cells (whose index of refraction is essentially that of water) and a stack of guanine crystals sandwiched between. Light is reflected from the interface at both sides of the stack of crystals. For certain wavelengths, constructive interference leads to an especially strong reflection.Bright light from a distant source is focused by the lens of a shark’s eye to a point on the retina, as shown in Figure V.2 c. The tapetum reflects these rays back through the lens, where refraction bends them into parallel rays traveling back toward the source of the light. Because the reflected light from the tapetum is directional, it is much brighter than the diffuse reflection from the shark’s body. But the bright reflection is seen by an observer—or a camera—only at or near the source of the flash that produced the reflection.Light of wavelength 600 nm in air passes into the layer of guanine crystals. What is the wavelength of the light in this layer?A. 1100 nmB. 600 nmC. 450 nmD. 330 nm
From the textbook chapter you will find a few key concepts needed to solve this.
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