Flow Cytometry Training Quiz

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Qallusions

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Quizzes Created: 1 | Total Attempts: 1,015

Questions: 13 | Attempts: 1,025 | Updated: Mar 21, 2023

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This quiz will test your knowledge of basic 4-color flow cytometry principles.

Questions and Answers

1.

What does light emitted as forward scatter (FSc) measure?
- A.
  
  Cell size
- B.
  
  Cell granularity/complexity
- C.
  
  Cell surface marker fluorescence
Correct Answer
A. Cell size

Explanation
Light emitted as forward scatter (FSc) measures the size of the cell. Forward scatter refers to the scattering of light in the forward direction when it encounters a cell. The size of the cell affects the amount of light that is scattered in the forward direction. Therefore, by measuring the forward scatter, we can determine the size of the cell.

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2.

What does light emitted as side scatter (SSc) measure?
- A.
  
  Cell size
- B.
  
  Cell granularity/complexity
- C.
  
  Cell surface marker fluorescence
Correct Answer
B. Cell granularity/complexity

Explanation
Light emitted as side scatter (SSc) measures the granularity or complexity of cells. When a laser beam passes through a cell, it scatters light in different directions. The amount of light scattered to the side is proportional to the granularity or complexity of the cell. Cells with more internal structures or granules will scatter more light, indicating higher side scatter values. Therefore, side scatter is used as a measure of cell granularity or complexity in flow cytometry and other techniques.

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3.

The FL1 channel measures emission in the green range. What fluorophores does channel 1 (FL1) measure? \ FL1 - Green emission (filter: 530/30)
- A.
  
  FITC, Alexa488, CFSE
- B.
  
  Phycoerythrin (PE)
- C.
  
  Allophycocyannin (APC)
Correct Answer
A. FITC, Alexa488, CFSE

Explanation
FL1 channel measures fluorophores that emit in the green range, which includes FITC, Alexa488, and CFSE. Phycoerythrin (PE) and Allophycocyannin (APC) are not measured by FL1 channel as they emit in different wavelengths.

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4.

The FL2 channel measures emission in the re/orange range. What fluorophores does channel 2 (FL2) measure? FL2 - Red/Orange emission (filter: 585/42)
- A.
  
  FITC, Alexa488, CFSE
- B.
  
  Phycoerythrin (PE)
- C.
  
  PerCP
Correct Answer
B. Phycoerythrin (PE)

Explanation
FL2 channel measures emission in the red/orange range, specifically with a filter of 585/42. Phycoerythrin (PE) is a fluorophore that emits in this range, making it the correct answer. FITC, Alexa488, and CFSE do not emit in the red/orange range, so they are not measured by FL2. PerCP is not mentioned in the question and therefore cannot be determined as a correct answer.

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5.

The FL3 channel measures emission in the red range. What fluorophores does channel 3 (FL3) measure? FL3 - Red emission (filter: 650 LP)
- A.
  
  Allophycocyannin (APC)
- B.
  
  PerCP
- C.
  
  FITC, Alexa488, CFSE
Correct Answer
B. PerCP

Explanation
FL3 measures emission in the red range, and PerCP is a fluorophore that emits in the red range. Therefore, FL3 measures PerCP.

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6.

The FL4 channel measures emission in the far red range. What fluorophores does channel 4 (FL4) measure? FL4 - Far Red emission (filter: - 661 nm)
- A.
  
  FITC, Alexa488, CFSE
- B.
  
  Allophycocyannin (APC)
- C.
  
  PerCP
Correct Answer
B. Allophycocyannin (APC)

Explanation
FL4 channel measures emission in the far red range, specifically at a wavelength of 661 nm. Among the given fluorophores, Allophycocyannin (APC) is the only one that emits in the far red range. FITC, Alexa488, and CFSE emit in the green range, while PerCP emits in the orange range. Therefore, the correct answer is Allophycocyannin (APC).

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7.

What is the order of brightest fluorophore to weakest fluorophore?
- A.
  
  FL1>FL2>FL3>FL4
- B.
  
  FL4>FL2>FL1>FL3
- C.
  
  FL2>FL3>FL4>FL1
Correct Answer
B. FL4>FL2>FL1>FL3

Explanation
The correct answer is FL4>FL2>FL1>FL3. This means that FL4 is the brightest fluorophore, followed by FL2, FL1, and FL3 in decreasing order of brightness.

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8.

What do you have to adjust to make sure emission of fluorescence from a single fluorophore does not bleed into multiple channels?
- A.
  
  Compensation
- B.
  
  Threshold
- C.
  
  Voltage
Correct Answer
A. Compensation

Explanation
To ensure that emission of fluorescence from a single fluorophore does not bleed into multiple channels, you need to adjust the compensation. Compensation is a technique used in flow cytometry to correct for spectral overlap between fluorochromes. By adjusting the compensation settings, you can effectively subtract the fluorescence signal from one channel that may be contributing to the signal in another channel. This helps to eliminate any cross-contamination between channels and ensures accurate and specific detection of each fluorophore.

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9.

What do you have to adjust to make sure different sized (extremely large or extremely small) cells can be visualized during acquisition?
- A.
  
  Compensation
- B.
  
  Threshold
- C.
  
  Voltage
Correct Answer
C. Voltage

Explanation
To visualize different sized cells during acquisition, you need to adjust the voltage. Voltage controls the intensity of the signal detected by the imaging system. By adjusting the voltage, you can optimize the signal-to-noise ratio and ensure that both extremely large and extremely small cells can be visualized clearly. Higher voltage settings may be required to visualize larger cells, while lower voltage settings may be needed for smaller cells to prevent signal saturation.

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10.

This setting has been previously adjusted on the machine to remove cellular debris from the graph settings during acquisition.
- A.
  
  Compensation
- B.
  
  Threshold
- C.
  
  Voltage
Correct Answer
B. Threshold

Explanation
The correct answer is Threshold. The explanation for this answer is that the setting mentioned in the statement, "to remove cellular debris from the graph settings during acquisition," suggests that the machine has a threshold setting. This setting allows the user to set a specific threshold value, above which any data or debris will be filtered out or ignored during the acquisition process. This helps to ensure that only relevant and accurate data is captured and analyzed.

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11.

What 2 lasers are included in the 4 color BD FACScalibur?
- A.
  
  An argon blue 488nm laser and a red 635 nm diode laser
- B.
  
  An argon red 488nm laser and a blue 635 nm diode laser
- C.
  
  A blue diode 488nm laser and a red 635nm argon laser
Correct Answer
A. An argon blue 488nm laser and a red 635 nm diode laser

Explanation
The correct answer is an argon blue 488nm laser and a red 635 nm diode laser. This is because the question specifically asks for the lasers included in the 4 color BD FACScalibur, and the argon blue 488nm laser and red 635 nm diode laser are the lasers included in this particular model.

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12.

Place these items in order from first to last for machine shut down. A) Make sure machine is in stand-by mode B) De-pressurize machine using black toggle C) insert tube of ddH2O and put holder all the way to the side for 1 minute D) insert tube of 10% bleach and put holder all the way to the side for 1 minute E) place holder under tube of bleach and leave for 5 minutes F) place holder under tube of ddH20 and leave for 5 minutes G) leave machine on for 15 minutes in standby mode to let lasers cool down H) remove protective shield from sheath fluid tank I) turn off FACScalibur J) replace protective shield on sheath fluid tank K) set machine to HI setting L) Empty waster container and fill sheath fluid tank
- A.
  
  A>D>E>C>F>K>B>H>L>J>G>I
- B.
  
  K>D>C>E>F>A>B>H>L>J>G>I
- C.
  
  K>D>E>C>F>A>B>H>L>J>G>I
Correct Answer
C. K>D>E>C>F>A>B>H>L>J>G>I
13.

What type of graph(s) is used to visualize cells during acquisition?
- A.
  
  Dot plot
- B.
  
  Histogram
- C.
  
  Dot plot and histogram
Correct Answer
C. Dot plot and histogram

Explanation
Dot plots and histograms are both types of graphs that can be used to visualize cells during acquisition. A dot plot is a simple graph that shows the individual data points as dots along a number line, allowing for easy identification of patterns and outliers. A histogram, on the other hand, displays the distribution of data by grouping it into intervals and representing the frequency or count of data points within each interval using bars. By using both dot plots and histograms, researchers can gain a comprehensive understanding of the characteristics and distribution of cells during acquisition.

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1

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Current Version
Mar 21, 2023

Quiz Edited by
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Mar 31, 2011

Quiz Created by
Qallusions

Flow Cytometry Training Quiz

What does light emitted as forward scatter (FSc) measure?

What does light emitted as side scatter (SSc) measure?

The FL1 channel measures emission in the green range. What fluorophores does channel 1 (FL1) measure? \ FL1 - Green emission (filter: 530/30)

The FL2 channel measures emission in the re/orange range. What fluorophores does channel 2 (FL2) measure? FL2 - Red/Orange emission (filter: 585/42)

The FL3 channel measures emission in the red range. What fluorophores does channel 3 (FL3) measure? FL3 - Red emission (filter: 650 LP)

The FL4 channel measures emission in the far red range. What fluorophores does channel 4 (FL4) measure? FL4 - Far Red emission (filter: - 661 nm)

What is the order of brightest fluorophore to weakest fluorophore?

What do you have to adjust to make sure emission of fluorescence from a single fluorophore does not bleed into multiple channels?

What do you have to adjust to make sure different sized (extremely large or extremely small) cells can be visualized during acquisition?

This setting has been previously adjusted on the machine to remove cellular debris from the graph settings during acquisition.

What 2 lasers are included in the 4 color BD FACScalibur?

What type of graph(s) is used to visualize cells during acquisition?