The basic unit of life is the cell, and now in a modern laboratory even mammalian cells can be easilycultured in a test tube or a flask. In this unit you have learned about the biomolecules of life such ascarbohydrates, lipids, proteins and nucleic acids which come together to allow cells to form andfunction. You are required to design a cell culture medium for maintaining mammalian cells, whatare your key considerations? (hint: you need to describe the key nutrients and the underlyingreason for them, and the pH of your medium).

Laboratory research and biotechnology require mammalian cell culture medium design. Several factors…

Answered: The basic unit of life is the cell, and…

Biomedical Instrumentation Systems

1st Edition

ISBN:9781133478294

Author:Chatterjee

Publisher:Chatterjee

Chapter24: New Technologies And Advances In Medical Instrumentation

Section: Chapter Questions

Problem 10P

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You are using fluorescence microscopy to study a plasma membrane protein that is fused to green fluorescent protein (GFP). Adding GFP to a protein allows us to monitor the protein in live cells using light microscopy without the need for any special stains. Under baseline conditions, the fusion protein is evenly distributed over the surface of the cell. You use the microscope software and laser to photobleach an ROI and watch for the recovery of fluorescence in real time. you have a control protein, a GFP fusion with the insulin receptor, where the ROI fully recovers from photobleaching in 10 min. However, when Protein X is fused with GFP, the fluorescence recovers only to 10% the starting levels in 10 min in the same cell type under the same experimental conditions. Provide two different, reasonable explanations for the different results with these two fusion proteins.
Explain (using the correct terminology and drawing an image) what happens to the cells in the following environments: A human blood cell (0.9% salt) is placed in a saltwater tank (27.5% salt). - A bacterial cell with 35% organic material in their cytoplasm is living in a tank filled with 35% sulfur. - A freshwater plant cell (75% water) is living in a pond that floods over.
A fluorescent molecule of 2000 daltons is microinjected into a single cell of an epithelial layer. When the layer is examined in a fluorescence microscope, the molecule remains confined to that cell. However, when a fluorescent molecule of 800 daltons is injected into a single cell of an epithelial layer, it quickly appears in the adjacent cells. Which of the following is responsible for the spread of the smaller molecule? 1. A) Desmosomes (maculae adherentes) 2. B) Focal contacts 3. C) Gap junctions 4. D) Intermediate junctions 5. E) Tight junctions (zonulae occludentes)
Which electron or light microscopy technique would you use for the following? (also explain) For visualizing the fine structure of cellular organelles and cell architecture in the cell interior using very thin cross-sections made in Epon. To obtain sequential images of a macromolecule in order to study its activity in real-time. To obtain an extremely clear three-dimensional image of the nucleus of a single cell. What is the magnification of the image in a microscope using a 20x ocular and a 100x objective?
In the context of cell biology, what do we mean by form follows function? What are a least two examples of this concept?
In a monolayer of cultured epithelial cells, the cells may or may not have formed tight junctions. What experimental test could be used to determine it the cells have functional tight junction structures? Place a small dye (1 k Da in molecular weight) into the culture media of cells cultured on a glass coverslip and use live imaging to monitor whether the dye can penetrate into the O cytoplasm of the cells, as would be expected if the cells have functional tight junctions. Inject a cell with a small dye (<1 kDa in molecular weight) and use live cell imaging to track whether the dye moves into neighboring cells, as would be expected if the cells have functional O tight junctions.
In your lab you are trying to culture skin tissue for use in transplant experiments. You’ve encountered a problem because the cells seem to dissociate after only one day in culture. What might be one solution to your problem (i.e., what might you add to your medium to get your cells to stick together)? Why?
List and describe the three main types of cytoskeleton. If you wanted to do immunocytochemistry to specifically stain each type of cytoskeleton, what is a protein that could be used for each cytoskeletal type (in other words, what is a unique protein for each cytoskeletal type)? What are three types of actin structures? Describe the involvement of actin structures in cell migration. How is the growth and shrinking of microtubules regulated? Then describe the roles of microtubules in: chromosomal separation during mitosis and the movement of organelles and vesicles within a cell. Describe a possible mechanism on how signaling pathways might impact the cytoskeleton, so that cell migration could be regulated in a localized manner within a multicellular organism. (hint: think about the possible transcriptional regulation of the G-protein regulators) What are 2 main challenges of protein targeting? Then describe one way these challenges are overcome during signal-based targeting and one way…
A patient has ingested a chemical known to prevent alpha-tubulin from hydrolyzing GTP to GDP. Which of the following is the BEST conclusion? O The patient's dyneins and kinesins will not be able to hydrolyze nucleotides. O The patient's microtubules will only be able to grow. O The patient's microtubules should still be able to grow and shrink depending on the needs of the cell. O The patient's microtubules will only be able to shrink.
The properties of integral membrane proteins can be studied by 1)Atomic force microscopy 2)Cryo-sectioning and electron microscopy 3)Freeze-fracture technique and electron microscopy 4)All of the above
The drug taxol is extracted from the bark of yew trees. It binds tightly to microtubules and stabilizes them. When added to cells, it causes much of the free tubulin to assemble into microtubules. Taxol can be used as an anticancer drug. At the molecular level, what does the drug prevent? At the cellular level, what part of cell division does it prevent? Be specific.
Which of the following statements are correct about how the complex biological matrix of the cell may affect protein folding, stability and function (select all that appy)? A. ATP tends to "bounce" along the surfaces of many proteins without forming a stable complex B. Proteins make up approximately 30% of the total volume of the cytoplasm in a typical cell C. Compettition between drug molecules and metabolites in the cytoplasm may cause unexpected drug side effects not seen with individual proteins in solution. D. Large protein complexes move faster through the cytoplasm than small metabolites. E. Complex intercations between proteins in the cell may significantly destabilize a protein compared to the isolated protein in water

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