Obtaining High Resolution Images And 3d Reconstruction

To make the specimen compatible with both forms of advanced microscopy, they sufficiently prepared samples by coupling the specimen with a fluorescence that was also conductive. This technique was accomplished with the FlouroNanogold label, which contains gold nanoparticles covalently bonded to a fluorescence label. That way, the LM worked as well as the EM for the same set of kinetochores that were being studied. The Hec1 protein was stained in this case because this protein naturally delineates the structures to be studied.

A light microscope was set up with the light on a low setting; one large Daphnia was selected and placed in the centre of a cavity slide by using a pipette.

Another limitation is out of focus light from outside the focal plane reducing image clarity or requiring constant adjusting.  On the other hand, under the right conditions, light microscopy allows for viewing of samples that are still alive.  There are aids such as fluorescent proteins, that can be used to track proteins in real-time in cells though the viewing of where the proteins are may not be very high in resolution, you may see roughly where the protein is in the cell, but will not be able to see the shape of the protein itself. Lastly confocal light microscopy gives moderately higher resolution, and significant enhancements in optical sectioning by limiting out-of-focus light.

The resolution in microscopy terms refers to the numerical aperture of the objective lens. The higher the numerical number the better the resolution of the image; also the shorter the wavelength the better the resolution (Alonzo p55).

Using the microscope provided to analyze the different size of the different cells and organisms

It was then during the 1950’s that Zacharias Jansen and his father, Han Jansen, created the first high-powered compound light microscope. Images could be magnified up to 9x which was a novel feat at that time. Zacharias and his father created the device after experimenting with glasses and discovered that by placing multiple lens in a tube they could increase magnification. Unfortunately, the microscope lacked the high resolution we have come to expect in modern-day light microscopes. The images produced by the Hans’ primitive microscope was extremely blurry and had limited

First, we discovered that we should start with low power objective to observe. To focus the image, use the coarse adjustment knob to adjust it. When looking at high power objectives, the fine adjustment knob can be used. Also we discovered that when you move the slide towards you, it appears to move away. When observing the letter “e”, we discovered that the images observed under the light are inverted and reversed. Although we could not easily tell with the feather, threads, and potato, it became noticeable with the letter “e”. The “e” was placed like “e”, however when looked into the eyepiece, the “e” was upside down. This shows that the microscopes works in an inverted way. We also discovered that to adjust the amount of light entering the microscope, we could use the iris lever to adjust the diaphragm. For example, the white thread required little light to see the cotton fibers, compared to the feather or letter “e”. Therefore, we learned that by altering the diaphragm, we can fix many of the problems associated with the observations. Lastly, we discovered that only one depth can be seen clearly at a time under high power. When working with the crossed strands of thread, we had to turn the fine wheel adjustment back and forth while looking through the microscope to focus one strand. All in all, the lab supported the purpose. We were able to identify,function the parts of a light microscope, and prepare a wet mount(of a feather, letter “e”, black and white thread, and a potato). Furthermore, we located objects using high and low power objectives, adjusted the diaphragm to attain correct lighting, and used stains for an easier and more detailed

The ways we can examine organism is by using a microscope. We’ve used a bright-Field microscopy that shines light through a specimen (1). The organism is then magnified as light refracts through the four objective lens 4X, 10X, 40X, and 100X found on the revolving nosepiece. It is then further magnified as we look through the ocular lens at 10X. By multiplying the objective lenses and the ocular lens gives a total magnification of 40X, 100X, 400X and 1000X. Once reaching objective lens 100X oil needs to be put on the slide. By using oil, it increases the lens magnification because oil refracts light just like the lens in the microscope and it has the same refractive index (1).

1.1: Super-resolution imaging of age-linked changes in actin cables and how deletion of Ykl affects those processes. We optimized a method to isolate yeast of different replicative ages and will use 2 different modes of super-resolution imaging to further characterize age-associated changes in the actin cytoskeleton. The resolution of conventional light microscopy (e.g. wide field and confocal) is diffraction limited, which restricts the ability of microscope to distinguish between two objects separated by a lateral distance less than approximately half the wavelength of excitation illumination. Structured illumination microscopy provides a 2-fold increase in axial (100 nm) and lateral (300 nm) resolution compared to

The VHX-6000 can create a three-dimensional image, by using a depth from defocus method by calculating height and analysing extremely small changes in texture. Even within areas of low contrast or under and over saturated pixels can be accurately calculated unlike when imaged using conventional microscope methods such

The purpose of this lab is to insure that all students understand how to correctly use the microscope. Students will all learn proper care and handling of the microscope, as well as learning the correct way to look at slides and specimens through the microscope.

The digital microscope Keyence VHX-5000 is a method, which is easy to handle and supports research with high-resolution images and the calculation of 3D-surface-models. Several functions make it easy and quick to work with: The 3D-function also allows clear 2D images of a rough surface, because all levels are focused automatically and a sharp image of all levels will be created. This function makes it easy to analyse deep cuts, which can be only partially focused in light microscopy or electron microscopy. To calculate a 3D- surface-model the lower and upper border of the depth of interest has to be determined, by focusing on these levels. Between these borders a level difference of five microns were chosen. In other words, after every scan the lens moves five microns from the sample away.

The session was very informative about how art can be use as a teaching tool in medicine. Before this session, I never thought anatomy images and diagnostic X-rays would be considered as artwork in medicine. I think the thought of strictly labeling certain observations as medicine could impede us from seeing beyond the image. Although the main purpose of X-ray imaging is to reveal the medical condition the patient has, it also holds a story of the patient. I have been told that having a good observing skill would be beneficial to physician-patient interaction but never thought this skill can be practiced through looking at arts. As we looking through a few paintings during this session, I started to have an idea of how this could be done.

NOTE: Answer Question A only if you used a compound light microscope for this experiment.

Two imaging systems have been tested, including an advanced X-ray imaging system that utilizes geometric magnification and a mobile phone imaging system. The aim of this experiment is to use resolution phantoms to determine system parameters including resolution, effective pixel sizes, magnification and other geometric parameters.