Blogs

The rhodamine labeled f-actin did not appear in the composite image of the three different fluorophores due to the intensity of the fluorescence being too low to register. This could be due to photobleaching that had previously occurred in this specific area of the sample, or if the sample was not labeled adequately with enough fluorescent dye. Time-lapse data for all three fluorophores under the same condition revealed discrepancies in rate of decay and initial intensity for each fluorophore. A relative high initial intensity for DAPI labeled dsDNA can be explained by the relative high net local concentration of bright fluorophores. Each nucleus contains a high concentration of dsDNA, which when stained with DAPI, creates a large solid fluorescent region with overlapping fluorophores. This differs from both tubulin and f-actin, which are of tube like nature, and appear as porous regions of interest where background light can seep through and be analyzed, making the initial brightness in the region of interest inherently darker. This difference is also reflected in the rates of decay, where in samples where there is less light to be diminished, the rate of decay appears much slower as it approaches the plateau of photobleached darkness. Time-lapse data for fluorescein under stained tubules under different conditions demonstrated discrepancies in rate of decay and initial intensity for all three conditions. The relative low rate of decay and low initial intensity for the images taken with a neutral density filters and auto shutter can be explained by the effect of the neutral density filter. The collective effect of the neutral density filters decreases the intensity of the epi-illumination light path by a factor of 32. This in turn lowers the ability for the fluorophores to be excited, causing a low initial intensity and low excitation, which leads to a lower rate of decay. The auto shutter condition displays a lower rate of decay compared to the open shutter condition due to the lower exposure to light over time. When the shutter is left on automatic, the sample is only exposed to light during the time an image is snapped as opposed to the entire duration of the time lapse. This lessens the time window that fluorophores can covalently bond to oxygen and other elements, and less brightness is lost over time. The open shutter condition leaves the sample exposed to the epi-illumination light path during the entirety of the time-lapse, allowing more time for covalent bonds to form with fluorophores and thus lose more brightness intensity over time. This explains the high relative rate of decay.  

Fluorescein labeled tubulin was then time-lapse imaged under several conditions: auto shutter, open shutter, and open shutter with neutral density filters. Data was analyzed graphically for all three conditions (Figure 3). Open shutter data was shown to have the highest initial intensity value and rate of decay (Table 2). Auto shutter data contained midrange values for both initial intensity and rate of decay (Table 2). Time lapse data with neutral density filters and an open shutter demonstrated the lowest values for initial intensity and rate of decay (Table 2). Selected time lapse images visually demonstrate the difference in excitation intensity over time during the photobleaching process under all three conditions (Figure 4).

When designing the figures associated with this plant, three images are used. First, the photo taken of the entire plant is labeled “A” in the top left corner of the photo. Next, the second photo used is the close-up of the flower. This photo is labeled “B” in the top left corner of the photo. Finally, the third figure was the geographical range of the plant. Owing to the fact that there is no clear range of this specific flower, the range of the three plants that “Blc Momilani ‘The Gypsy’” is a hybrid of are used to design the map.

Starting with a blank world map, the range of Brassavola, the first plant, is added. Brassavola range includes Mexico, the West Indies, and Central America, so the areas are circled in black and then lightly filled with green, making it possible to slightly see the world map beneath. Next, the second plant, Cattleya’s range is added to the map. Cattleya’s range includes Costa Rica to Argentina, so that area is circled in black and lightly filled with red. Finally, the last plant, Laelia, has a range that spans from Western Mexico to Southern Bolivia. This area is circled in black and then lightly filled with purple. In some areas of the map, these colors overlap, so the colors that overlap are arranged in stripes in any areas where this happens. Once the world map is complete, it is labeled with a “C” in the top left of the picture. Figure A and B are arranged next to each other, with C arranged directly below them.

According to a recent publication, Microorganisms found in dirt can yield antibiotics that can kill pathogens that already have found a way to create drug-resistance to known antibiotics. Soil samples across the country were analyzed, particularly in the search for known genetic sequences used in the production of antibiotics. A sequence was identified and a new possible antibiotic has been tested on the skin of rats which showed signs of sterilization of MRSA. Even if this method initially fails to kill pathogens in human cells, the discovery brings hope as increasing antibiotic resistance is becoming becoming a huge problem all around the world. 

This experiment shows that the diet of the birds is what most likely caused their mercury levels to be so elevated, so the next part of their experiment was to study the mercury levels in the bird’s main prey. This part of the experiment showed that the prey did have elevated mercury, but it further proved that the higher in the food chain, the more mercury in the system. In this case, spiders had a much higher mercury level because it eats other organisms that already have mercury in them; and if spiders have this much more mercury, then birds would be significantly higher because they are so far up the food chain. 

I had my third intramural basketball game today. I've been extremely sick for the past 3 days and I didn't think I could make it. But because we were 2-0 in the season, I wanted to go support the team by playing. I ended up playing most of the game forgetting that I was sick because I was way too into the game as my adrenaline was through the roof after taking an energy drink. I hurt my arm but I didn't even realize it hurt a lot until after the game was over. Sadly, we ended up losing the game but it was an eventful night and it was fun to play some basketball with my fraternity brothers. It is crazy how human body works when the adrenaline kicks in.

Two pictures are taken: a close-up picture of an individual flower and a picture of the entire plant. The close-up of the individual flower includes a fully blossomed flower. The fully blossomed flower is pink in color and has multiple layers of petals. Measurements of the flower size is taken with a ruler in centimeters and is recorded for future reference. The picture of the entire plant is taken from a distance at an angle in which the plant is in between the entrance and the camera. The size of the entire plant is estimated by standing next to the plant. Along with the two pictures obtained from the Durfee Conservatory & Garden, is a world map showing the origins of the Camellia Japonica Napoleon plant. This information is found by researching the origins of the Camellia Japonica family, rather than the specific individual Napoleon species. The countries where these plants grow in the wild is marked on the blank image of a world map.

The species of plant that is being observed is the Camellia Japonica Napoleon. This plant is located at the University of Massachusetts Amherst in the Durfee Conservatory & Garden. The greenhouse is in between the morrill science buildings and the university health center. Facing the morrill science buildings is an entrance to the greenhouse, where the different kinds of Camellia Japonica are found on the right immediately after entering the greenhouse. The Camellia Japonica Napoleon is the second to last tree from the entrance.

The plant was located by entering the Durfee Conservatory via the West entrance. The plant was the closest in the left row along the aisle. To photograph the whole plant, the photographer stood by the entrance door and aimed his phone camera at the plant. He then walked backwards towards the right of the building until the whole plant could fit in the frame of the picture. The close-up picture of the flower was taken of the closest flower to the door along the aisle side of the plant. The phone was moved close to the flower in order to frame the flower in the image and the photo was taken. To provide a scale, another picture was taken with a “Dunkin Donuts” gift card held above the flower. The name of the plant was taken from a card on the pillar from which the plant was growing.

Introduction

             As a beginning, I wanted to get the images from the Ray Ethan Torrey Botanical Collection at the Morrill Greenhouses being that I have passed by a countless number of times in the past two and a half years. Being from a part of Central America, I was determined to get a plant that was native to either Central America or the upper regions of South America. In addition, I want a plant that is distinct in that it contains few flowers with distinct features as oppose to a plant with multiple flowers surrounding the plant itself, which would make it difficult to pinpoint the specific flower that will be individually photographed. A plant potted on the floor or on a shelf mid-height will enable others with shorter statures to have easy access over high-hanging ceiling or wall plants.

The botanical collection will be best illuminated at around mid-day when the sun is the highest entering the windows on the top of the greenhouses. Both images of my figure will be taken at the same time to minimize discrepancies between them as well as same height, although different angles will be needed to get a more in-depth view of the plant. Both images taken on a vertical axis as oppose to horizontal. The range figure will be taken off a website where marking the range is simple with little no to chance of error.

The images itself will be labeled A, B and C on the upper left-hand corner of each. The multi-panel figure itself being build using an online source with the images of the flowers itself above and below one another with the close-up being below the further image. These two will be on the left of the range map which will serve the same height of both plant images to make a larger square.