User:Brian P. Josey/Notebook/2009/08/27: Difference between revisions
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On Andy's suggestion, I spent the first part of the day reviewing basic microscopy. The things that I looked at included Koehler illumination, bright field, DIC and fluorescence. I also looked up the basic components of a microscope. They are: | On Andy's suggestion, I spent the first part of the day reviewing basic microscopy. The things that I looked at included Koehler illumination, bright field, DIC and fluorescence. I also looked up the basic components of a microscope. They are: | ||
*Eyepiece- Two or more lenses that you look into to see the image | *Eyepiece- Two or more lenses that you look into to see the image. | ||
*Objective lenses- One or more lenses that collect light from the sample | *Objective lenses- One or more lenses that collect light from the sample. | ||
*Stage- A platform used to hold the sample. | *Stage- A platform used to hold the sample. | ||
*Illumination source- Simply gives light to the microscope. | *Illumination source- Simply gives light to the microscope. | ||
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* Differential Interference Contrast (DIC)- The two microscopes that are used in the lab use DIC, which relies on polarized light to create a finer image, based on the optical properties of the sample. The basic path of the light is: | * Differential Interference Contrast (DIC)- The two microscopes that are used in the lab use DIC, which relies on polarized light to create a finer image, based on the optical properties of the sample. The basic path of the light is: | ||
**Unpolarized light enters the microscope and is polarized at 45 degrees. | **Unpolarized light enters the microscope and is polarized at 45 degrees. | ||
**The light then enters a Wollastom prism and is split into two | **The light then enters a Wollastom prism and is split into two sets of waves that are polarized to each other at 90 degrees. These two set of waves are the sampling and reference rays. | ||
**The rays are focused by a condenser to pass through a sample. When they do pass through the sample, two corresponding waves are offset from each other by about 0.2μm. | **The rays are focused by a condenser to pass through a sample. When they do pass through the sample, two corresponding waves are offset from each other by about 0.2μm. | ||
**They pass through the sample at two different, adjacent areas of the sample. This in essence creates two adjacent bright field images that are slightly offset from each other. | **They pass through the sample at two different, adjacent areas of the sample. This in essence creates two adjacent bright field images that are slightly offset from each other. | ||
**The light then travels through the objective lens and focused | **The light then travels through the objective lens and focused before entering a second Wollaston prism. | ||
**In the prism, the light is recombined at a polarization at 135 degrees. This creates interference patterns, lightening and darkening the images at points corresponding to the different optical qualities of the sample. | **In the prism, the light is recombined at a polarization at 135 degrees. This creates interference patterns, lightening and darkening the images at points corresponding to the different optical qualities of the sample. | ||
*Fluorescence Microscope- A fluorescence microscope uses fluorescence, and phosphorescence instead of or in addition to reflection and absorption. To do this, fluorescent molecules are attached to the subject. A particular wavelength of light is filtered out from a light source and then falls on the sample. The fluorescent molecules then absorb the light, and release light with a longer wavelength. This new set of light is then collected in the same way that all light is collected, creating an image of the sample. | *Fluorescence Microscope- A fluorescence microscope uses fluorescence, and phosphorescence instead of or in addition to reflection and absorption. To do this, fluorescent molecules are attached to the subject. A particular wavelength of light is filtered out from a light source and then falls on the sample. The fluorescent molecules then absorb the light, and release light with a longer wavelength. This new set of light is then collected in the same way that all light is collected, creating an image of the sample. |
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Notes on MicroscopyOn Andy's suggestion, I spent the first part of the day reviewing basic microscopy. The things that I looked at included Koehler illumination, bright field, DIC and fluorescence. I also looked up the basic components of a microscope. They are:
The basic techniques and types of microscopes are:
After looking up these things I began to read "Mechanical Design of Translocating Motor Proteins" by W. Hwang. It is a review paper on how both kinesin and myosin, among other motor proteins work. Notes on this paper will come in a later entry. |