Cell Biology Chapter 11 & 14 Quiz

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Questions and Answers

What is the principle behind ion-exchange chromatography?

It relies on the temperature of the solution.

It uses the charge of proteins to separate them. (correct)

It separates proteins based on their size.

It separates proteins based on their affinity to a specific substrate.

What is a primary advantage of using TIRF microscopy during imaging?

It allows for deeper tissue penetration.

It minimizes background light by illuminating only a thin layer. (correct)

It uses higher-energy light to enhance imaging speed.

It can illuminate the entire sample at once.

How does confocal microscopy improve the clarity of images?

By employing a pinhole to focus on a specific plane. (correct)

By cooling the sample to reduce movement.

By using fluorescence from all layers of the sample.

By using multiple color filters.

What is a key benefit of using multi-photon microscopy compared to traditional confocal microscopy?

It facilitates deeper imaging with less damage. Signup and view all the answers

What does GFP technology allow researchers like Dr. Smith to do in their experiments?

Track the dynamics of proteins in live cells. Signup and view all the answers

How does multi-photon microscopy reduce damage during imaging?

By utilizing lower-energy infrared light. Signup and view all the answers

What happens to proteins during affinity chromatography?

Target proteins bind to specific molecules in the column. Signup and view all the answers

Which of the following is a common issue Dr. Smith needs to address during his imaging process?

Background noise affecting image clarity. Signup and view all the answers

What is the main advantage of using size-exclusion chromatography in protein isolation?

It separates proteins on the basis of their size. Signup and view all the answers

In FRET analysis, what is indicated by a decrease in fluorescence from one protein and an increase in fluorescence from another?

The proteins are interacting closely. Signup and view all the answers

What conclusion can be drawn if a researcher observes rapid fluorescence recovery after bleaching in FRAP experiments?

The proteins are highly mobile. Signup and view all the answers

How does super-resolution microscopy like STORM improve imaging over conventional microscopy?

It achieves resolutions at the nanometer scale. Signup and view all the answers

What is the purpose of differential centrifugation in protein isolation?

To collect the supernatant containing soluble proteins. Signup and view all the answers

What role does ion-exchange chromatography play in protein purification?

It separates proteins based on their charge. Signup and view all the answers

Which microscopy technique allows for the visualization of specific proteins using a fluorescent tag like GFP?

Confocal microscopy. Signup and view all the answers

What is a key feature of TIRF microscopy compared to traditional optical microscopy?

It only illuminates a thin region of the sample. Signup and view all the answers

How does gel filtration chromatography separate proteins?

By size, using beads with tiny holes Signup and view all the answers

What is a primary benefit of using confocal microscopy?

It only captures light from the focal plane, reducing blur Signup and view all the answers

What distinguishes differential centrifugation from density gradient centrifugation?

Differential centrifugation spins extracts at increasing speeds Signup and view all the answers

Which of the following is true regarding multi-photon imaging?

It allows deeper tissue penetration with less damage Signup and view all the answers

What role does electrophoresis serve in protein analysis?

It visualizes proteins while confirming size and purity Signup and view all the answers

How does TIRF microscopy differentiate from standard fluorescence microscopy?

It excites only the fluorescent molecules close to the surface Signup and view all the answers

What is one of the main applications of GFP technology in research?

For real-time visualization of protein localization Signup and view all the answers

What advantage does density gradient centrifugation offer for separating cell components?

It separates components based on their varying densities Signup and view all the answers

Study Notes

Week 8 (Oct 24): Mitochondria & Chloroplasts – Structure

Chapter 11: pages 637-651, 653 (ion channels) - 661
Chapter 14: pages 811-817, 861-868
Chapter 12: pages 726-735
Chapter 14: pages 843-844

Confocal Laser-Scanning Microscopy

Confocal laser scanning only allows light from the exact focal plane to be captured
This prevents light from areas above or below the focal point, reducing blurry images.
Electrophoresis is used to separate proteins based on size and charge
SDS protein gives a uniform negative charge, making proteins linear
This allows visualization for protein size and purity
Confirm correct separation for further studies or applications

Cell Fractionation

Cell fractionation breaks open cells to release contents (proteins, organelles) to create a cell extract
The cell extract allows separation of cellular components
Centrifugation and chromatography are used in protein purification
These methods separate proteins by size, charge, or other properties

Differential vs. Density Gradient Centrifugation

Feature	Differential Centrifugation	Density Gradient Centrifugation
Method	Spins extract at increasing speeds	Uses a sucrose gradient
Separation	Heavier parts settle first (nuclei)	Components move at different speeds, forming distinct bands
Basis	Looks at the mass of components	Looks at the size of components

Column Chromatography

Gel filtration: Separates proteins by size. Larger proteins pass through holes faster than smaller ones.
Ion exchange: Separates proteins by charge. Proteins with opposite charge are attracted to the column, separating them by changing salt or pH.
Affinity: Separates proteins based on binding to specific molecules. The column binds to proteins of interest. Other molecules pass through without binding.

Super-Resolution Techniques

STORM: Enhanced imaging capabilities compared to conventional microscopy,
Allows scientists to see details at the nanometer scale (smaller than conventional light microscopes)

Protein Isolation Methods

Differential centrifugation: Separates cellular components by size and density.
Size-exclusion chromatography: Separates proteins based on size. Larger proteins elute first.
Ion-exchange chromatography: Separates proteins based on charge.
SDS-PAGE: Separates proteins based on size after electrophoresis. SDS gives proteins a uniform negative charge, denaturing them.

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Description

Test your knowledge on mitochondria and chloroplasts as covered in Chapters 11 and 14. This quiz focuses on the structures, functions, and methodologies such as confocal laser-scanning microscopy and cell fractionation. Review your understanding of protein separation techniques and cellular components.