The Wild Cherry/Sweet Cherry Tree

 

Prunus avium, commonly known as the wild cherry or sweet cherry, is a species of cherry tree native to regions within Europe, Anatolia, Maghreb, and western Asia. The species is widely cultivated, and has become naturalized in both North America and Australia.

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A widefield sample of wild cheery bark. Notice the different cell sizes change over a given area.

The wild cherry is one of two species of cherry that supply the world’s cultivators of edible cherries.

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10x magnified area of the sample. Here complex structures are more visible, giving a more in depth look into the cells.
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Interesting components are sometimes present within the sample. This foreign structure appears to have become part of the sample.
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Different cells make up the sample of the wild cherry tree. Each cell has a given purpose to maintain health and continued growth.

The tree is also cultivated as timber, and is a highly valued hardwood that is used for woodturning, to create furniture cabinets and musical instruments. The wood is also used for the smoking of meats in North America, giving a distinctly pleasant flavor to both pork and poultry.

Color In Microscopy

Microscopy allows people to view the world on a significantly smaller scale, and although many of things we see are dyed by people, there are still many subjects that are not or simply cannot be.

There are some subjects that, under certain methods and conditions, can actually show immense amounts of different color without having to dye it. I selected three methods that I captured in these photomicrographs to display just a small idea of what colors can be found.

Each method has their own way to “extract” these colors, and sometimes it can produce more varieties of color paterns than just what’s shown for the same subject.

The three sets of images use fluorescence, differential interference contrast, and cross polarization, respectively. Each of the methods rely heavily on what the subject is to exibit the different colors shown in the image. Some subjects, such as autofluorescent organisms, will have more profound colors than others.

These images show that color can be found in even the smallest of subjects, it’s simply a matter of just finding what method exploits these colors.

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Sambucus Lenticil at 10x magnification. Left image is from blue excitation fluorescence and right is from green excitation fluorescence.

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Silicon semiconductor wafer at 10x magnification. Image was taken using the differential interference contrast method.

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Dyed hair at 10x magnification. Image was taken using the cross polarization method.

Darkfield Illumination

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Darkfield illumination is one of the various methods of lighting used in microscopy to capture the small, and make it large. This method illuminates the subject through the use of oblique angle lighting to create a glowing and unique presentation of a subject. Oblique lighting eliminates the zero order of illumination which is what sets this method apart from Brightfield illumination. The visible light consists of rays refracted from the subject into the imaging system.

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In order to create Darkfield illumination, a microscopist requires a Brightfield objective and a Darkfield condenser. When executing Darkfield, it is important to have the disc stop equal to the numerical aperture of the objective at the location of the aperture diaphragm exit pupil. Additionally, one must open the field stop fully to ensure the the light is unfocused. Then the operator will raise and lower the substage condenser until the apex of illumination reaches the sample plane.

IMG_9089Advantages of Darkfield Illiminated Photomicrographs

  • Effective for semi transparent subjects
  • Useful for delineating edges of subjects
  • Can reveal internal structures not seen with Brightfield illumination

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Botanical Samples

Botanical samples tend to, on average, have an increased width compared to human specimens due to the differing cutting techniques used. Observing botanical samples through the use of photomicrography allows researchers to observe the development of plants. There are a plethora of plant species, and all of them grow differently. This differing growth can be observed through cross sections of plants through the microscope and microphotography. Through various techniques, distinct features of plants can be illuminated and brought forth. Two examples of these lighting set-ups are darkfield (Fig. 2) and fluorescence (Fig. 3). The darkfield lighting technique uses scattered luminance to light only the subject, therefore producing a dark background and bright subject matter. The fluorescent method takes advantage of the naturally occurring fluorescent aspects of the plant life, thus lighting up the parts of the sample that reflect this specific area of the spectra.

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Figure 1: The image above visualizes normal Kohler Illumination.

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Figure 2: This image shows the darkfield technique.

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Figure 3: The image above illustrates fluorescence.

Wide Field Imaging

Kugler_20160229_IMG_0814 PanoramaThe image you see in the above was taken by a procedure called Wide Field Imaging. Wide Field Imaging can be accomplished by taking several images of a subject of your own choice. This is generally done to obtain high amounts of detail and the whole subject at the same time. Many photographers tend to use 10x objectives for their choice of magnification so that the images taken say around 100, causing the final compile to have a smaller file size. After obtaining the images, there are several ways to process them, Photoshop, PTGui, or any other panoramic photo stacking application. For the best results have a 50% overlap in subject matter between each image.Kugler_20160229_IMG_0907

The subject is a Nymphaea water lily. The piece above is a Sclereid. It is a type of stone cell. They help support and conduct water through out the plant.

The piece below is a vein. Unlike humans, it doesn’t carry blood. Instead it transports water and food to keep the plant well nourished.Kugler_20160229_IMG_0911-4 copy

Human Spinal Cord

Human Spinal Cord

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The spinal cord is a long, thin, tubular bundle of nervous tissue and support cellsthat extends from the medulla oblongata in the brainstem to the lumbar region of thevertebral column. The brain and spinal cord together make up the central nervous system (CNS).

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Posterior Gray Horn

the three membranes that envelop the brain and spinal cord with the primary function of the meninges is to protect the central nervous system.

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Meninges

the three membranes that envelop the brain and spinal cord with the primary function of the meninges is to protect the central nervous system.

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Posterior Gray Commisure

The gray commissure is a thin strip of gray matter that surrounds the central canalof the spinal cord and, along with the anterior white commissure, connects the two halves of the cord. It comprises lamina X in the Rexed classification.

Human Scalp

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The structure of the scalp can be broken down into five layers. The human scalp is covered with one of the thickest sections of skin on the body. Its largest thickness is located at the back of the head and as it reaches the front it becomes thinner. The skin of the head contains numerous sebaceous gland and hair follicles; they all produce oil.Beneath the surface is a thin connective tissue layer called the superficial fascia. The connective tissue is a dense layer of fats and fibrous tissue that contain the nerves and vessels of the scalp. Next is the epicranial aponeurosis which is a tough layer of dense fibrous tissue. Following is a loose areolar connective tissue layer that provides a separation between the upper three layers and the paricranium. The paricranium is the membrane that covers the outer surface of the bone and provides the nutrients needed for the bone as well as for repair.

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The Bigger Picture

Wide field imagery in Microscopy takes the smallest of objects and presents them in a grandiose way. This methods purpose is to be able to display the entirety of a sample for reasoning’s of research or demonstrations.

The process of making one of these images usually takes a lot more than just one picture. For me it was generally in the 200 range. This is because the images have to be at a high enough magnification to delineate small structures within a sample, such as a nucleus within a cell. By doing that, however, you cannot get the entire subject within the photo. This process for me usually is an hour long session slouched over a camera making small adjustments after each photo.

After the initial photographing of the images comes a session of   staring at a computer screen, to stitch all the photos. This part can sometimes be fairly tedious. To begin this section of the process you need to load all your images into a stitching software, which there are several. Next you wait anxiously to see if the software will actually stich the photos. Depending on well you shot this can go either way. When the software finally spits out a correctly stitched image of your sample and not a cubist representation of it you can sit back in your chair and cheer a little.

To finish your image you need to add an appropriate bar scale to show the size of you subject. Then if you’d like you can process it a bit to make it look pretty. Yeah, that’s essentially it in a nutshell. If you follow most of these steps you should hopefully have a presentable wide field photomicrograph.

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Structures of the Human Body

The lens can be seen in the bottom right of the image of the eye of the human fetus. The lens, along with the cornea, allows light to be focused on the retina. The retina is located at the back of the eye. The suspensory ligament can be seen left of the lens. This structure helps to keep the lens in place, and is located on both sides of it. The middle picture is a bone marrow sample. Bone marrow is the spongy material inside bones. Bone marrow has stem cells, which produce blood cells. Red blood cells, platelets, and white blood cells are all examples of blood cells that are located throughout the body. The bottom picture is a portion of the heart wall. The heart is the strongest muscle in the human body. Veins carry oxygenated blood towards the heart, while arteries carry deoxygenated blood away from the heart.

Wide Field Photomicrography

To create wide field images there are simple steps to follow. First, set the camera to raw format in the settings. Next, go pick out a slide or two and attach the camera to the microscope. Find the specimen and focus the image for the camera. Start on one side of the slide and work across, leaving about ¼ of the previous image in the next. Continue to work across and down until the end of the specimen. Take the SD card out of the camera and put it into the computer. Pull the photos into Photoshop and automatically Adobe Camera Raw will pop up. Here, make some minor changes throughout the images and save the images as JPEGs. Next, go into Adobe Bridge and select the best images of each section of the specimen that can be combined. Choose about 5-10 images, hover over the tools drop down menu and hover over Photoshop and click photomerge. Automatically the program will go into Photoshop and then select reposition for the images chosen. Hit done and the section selected will be made. Do the same thing for the rest of the images. By the end there should be multiple sections about 5-6 depending on how many primary images. Next, flatten the layers in all the selection photographs and save them as sets of JPEGs. Now go back to Adobe Bridge and select the section images and repeat what was done to make the section images. Make a bar scale of 100 microns. Lastly, give the image a title, the name of the slide, and save the whole image as a TIFF file.

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