in flow cytometry, when cells labeled with fluorescent molecules pass through the focused laser beam, what two types of light signals are generated?
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in flow cytometry, when cells labeled with fluorescent molecules pass through the focused laser beam, what two types of light signals are generated?
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- 1) If direct stains work so well, why would we ever use indirect stains? Give two reasons why you might choose to use indirect stains instead of direct stains? 2) Assume that two days (dye A and dye B) were applied to bacterial smear. The cells were stained by dye A while the slide background was stained with dye B. Which stain had an anionic chromogen? Which stain had a cationic chromogen?The cell is usually placed after the monochromator, except in the case of using a multichannel detector. Why?Describe what is gel electrophoresis?
- a. What can you observe in viewing your stained bacterial slide under the microscope if you fixed a lot of bacterial colonies in your slide during smear preparation? b. What can you observe in viewing your stained bacterial slide under the microscope if you heat fixed your slide in a much longer exposure to heat?A flow cytometer is constructed of many components. For good quantitative results how and how frequently should the various systems be calibrated? What is MESF and how does it apply in flow cytometry?Assume you are observing the diatom pictured in Figure 1 using the 10X lens in a compound light microscope. You move to the 40X lens and then again to the 100X lens by only rotating the turret (remember that the lenses are parfocal), without making any other adjustments to the microscope. c) After making your adjustments, you notice that the midline of the diatom is in focus while the remainder is blurry. Explain, based on microscopy principles, why this has occurred Provide a description of the procedure to prepare an acidic stain of bacteria using Nigrosin as it would appear in the Methods section.
- How do you figure out a number of live cells using flow cytometry results?Ms. Michi is a researcher in a renowned pharmaceutical company based in Singapore. One of the cell lines in the lab is suspected to be contaminated. She is culturing NIH 3T3 mouse embryo fibroblast cell line (NIH/3T3 (ATCC® CRL-1658TM). Figure 1 below illustrates her observation under the inverted phase contrast microscope. a) b) c) d) e) Figure 1 (Routray et al., 2016) What type of contamination is her laboratory facing? Explain your answer. Give FOUR (4) suggestions to Michi in order to prevent such contamination in future Besides the contamination stated in Question 2 a), name FIVE (5) other contaminations that Michi should be aware of. During a routine check, Michi's superior requests her to characterize cell lines available in the storage to determine their authenticity. Describe TWO (2) effective cell line characterization methods that she can utilize for this purpose. The pharmaceutical company wants to improve its toxicity testing by replacing monolayer culture with spheroid…draw the results of a flow cytometry experiment in which one set of cells is treated with vehicle and the other set is treated with Taxol.
- A student has a compound microscope equipped with 10X ocular lenses and 4X, 10X, 40X and 100X objective lenses. The student has a slide with cells that are -100 microns (1 micron = 103 mm) in diameter, and wants to view a magnified image of a %3D cell so that the cell appears to be 10 mm in diameter. Which objective lens should the student use? O 10X O 40X O 100X O 4X Question 7 2 ptsExplain the differences between gel electrophoresis and column chromatography. Address the principles behind each separation. Why do large molecules migrate more easily in one method and with more difficulty in the other? Which method generated the most precise results?You got an opportunity to join a professor lab who is working in-vivo model and specifically looking at the dysregulation of mitochondria in liver. He asked you to isolate mitochondria from a Rat liver and placed in an assay medium. Based on the knowledge you gain in this course so far, please answer the following questions: a) Which technique will you use to isolate mitochondria? b) What happens to the pH of the medium when the medium is kept anaerobic? c) What happens when O2-saturated saline is added to the mixture?