Cell Biology Lab 1: Microscopy & Characterization of CellsField size at each magnification. 4x = 5 mm10x = (4/10)*5 = 2 mm40x = (4/40)*5 = 0.5 mm100x = (4/100)*5 = 0.2 mm Summary and Observation of samples A-E.A) Red Blood Cells 10x- Very small dots (circular) in a muted red colour were visible through the entire field of view.40x- Muted/light red (circular) dots visible at a large scale, less in the field of view.Oil Immersion- A single muted/light red (circular) dot visible in the field of view, estimated size around 0.
015 mm.B) Yeast 10x- Particles visible look vaguely like a fuzzy green substance, almost like dust.40x- Cells look like yellow tinted scales across the field of view.Oil Immersion- Oval in shape and light yellow-green in colour, some stagnant and some are moving throughout the field of view. Estimated size around 0.02 mm.C) Bacteria 10x- Particles look like dust.40x- Unable to view anything.
Oil Immersion- Unable to view anything. D) Algae 10x- Two jagged edged cells visible in the field of view.40x- One jagged edged cell visible in the field of view.Oil Immersion- A long strand that stretched out along the field of view (Approximately 0.2 mm in length and 0.03 mm in width), green around the edges and purple in small cracks along the center.E) Cheek Swab (Cassie) 10x- Unable to find anything at this magnification without staining. After staining, three cells visible, red in hue, scaly/ridged texture. Estimated size around 0.4 mm.40x- An uneven, blob like shape visible in the field of view, green-blue in hue.Oil Immersion- (After staining with Safranin) Cell visible in field of view, red tinged, long and bubbly in shape with a ridged pattern/texture. The Red blood cells and the Yeast cells were close in size and didn’t greatly differ in shape though they differed in colour and overall structure. The Algae cells were much larger in length than any of the other cells were and had a mixture of green and purple in colour. The Cheek cells were the largest in width of the cells viewed and had an interesting shape and texture to them that wasn’t visible on any of the other cells. We were unable to view out bacteria sample and think that heat fixing may have helped.Question: What else could have been on the cheek swab slide.Proposed technique: Gram Staining.The paper that I found isn’t directly related to the cells and bacteria within the human oral cavity but rather that of snakes from Southern Africa and how the bacteria found within the oral cavity effects snake bite victims. They used seven different species of snake and took oral swabs looking for bacteria, they prepared smears of these samples and gram stained them in order to look at the type of bacteria within each sample. Out of the 15 bacterial species they were able to isolate they found that 20% of the species were gram positive aerobic cocci (Blaylock, 2001). They used the information isolated from these tests and related them back to samples taken from snake bite wounds to determine the correlation between the infection that occurs. Though this article wasn’t directly related to our lab it was still an interesting read.Reference:Blaylock, R.S.M., (2001), Normal oral bacterial flora from some southern African snakes. Onderstepoort Journal of Veterinary Research. 68:175-182.3. Cells ranked in order of size, Largest to Smallest.1) Cheek Swab (0.4 mm)Field of view at 10x mag (2 mm)2 mm / 5 (times cell could fit across field) = 0.4 mm2) Red Blood Cells (0.015 mm)Field of view at 100x mag (0.2 mm)0.2 mm / 13 (times cell could fit across field) = 0.015 mm3) Yeast (0.02 mm)Field of view at 100x mag (0.2 mm)0.2 mm / 10 (times cell could fit across field) = 0.02 mm4) Algae Cell (Length- 0.2 mm , Width- 0.03 mm)Field of view at 100x mag (0.2 mm)0.2 mm / 6 (times cell could fit across field using Width) = 0.03 mm0.2 mm / 1 (times cell could fit across field using Length) = 0.2 mm5) Bacteria- We were unable to view anything for this sample. 4. Scientific Drawing.I chose the Algae cell to draw because it was by far the largest (When referring to length of the cell) and most clear image we obtained with our wet mounts. It was strange for me to see an algal cell look like this since I had only ever seen rounded algal cells before and I still question whether what we saw and what I have drawn here is in fact an algae cell.5a. Comparison of cultured cells to Cheek and Red Blood Cells. The cultured cells were much smaller and harder to view than the cheek cells and the red blood cells were where we were able to view those cells much quicker under the microscope, they were easier to find. The cheek and red blood cell samples were also much easier to view at a lower power, whereas the other samples were only truly easily visible and able to be described under oil immersion.5b. Cultured cells as Experimental Model. In the paper I chose they use epidermal cells from humans and cultured those cells to produce an epithelium layer they propose could be used to aid in the repair of damaged epidermis in a patient for medical purposes, such as skin grafting for burn victims. The cells were used as they were relatively fast to culture and were able to form a sturdy layer that could be lifted from its medium. Though the cells wouldn’t be exact to the epidermal layer of the patient, the epithelium would be derived from a sample of their skin and work as a relatively close alternative. In this case the cells resemble those growing in the human body since they are derived directly from the patient’s cells or samples given by other humans, however they wouldn’t be quite as structurally sound as the normal epidermal layer. The produced layer would be lacking the pigmentation of the original cells and the ability to harbor normal functions such as adapting to sweat glands.Reference:Green, H., Kehinde, O., & Thomas, J. (1979). Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. Proceedings of the National Academy of Sciences, 76(11), 5665-5668. doi:10.1073/pnas.76.11.5665 6. Description and use of 2 types of microscopes. Fluorescence microscopes- Used in conjunction with fluorescent tagged cells in order to get a better and more detailed view of the cells components.Electron Microscopes- Give the best resolution but are unable to be used for looking at living cells. There are Transmission Electron microscopes and Scanning Electron microscopes which use electrons instead of light in order to view the sample.Reference:Alberts, Bray, Hopkin, Johnson, Lewis, Raff, . . . Walter. (2013). Essential Cell Biology (4th ed.). New York, NY: Garland Science, Taylor & Francis Group. 7. Cells viewed under inverted epifluorescence microscope and fluorescence explanation.The cells viewed under the inverted epifluorescence microscope fluoresced green in colour and were almost like scales or tiny circular cells in multiple rows across the entire field of view. The sample looked like a smear almost.The cells would have been tagged with a fluorescent dye that is excited by the light emitted from the microscope and allow them to properly fluoresce. Different colour dyes can be used to tag/label specific parts of the cells that you want to look at. Reference:Alberts, Bray, Hopkin, Johnson, Lewis, Raff, . . . Walter. (2013). Essential Cell Biology (4th ed.). New York, NY: Garland Science, Taylor & Francis Group.
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