1.
Traditional flow cytometry is useful for:
Correct Answer
A. Discriminating cells based on changes in intensity
Explanation
Traditional flow cytometry is a technique that uses laser-based technology to analyze and sort cells based on various characteristics. One of the main advantages of flow cytometry is its ability to discriminate cells based on changes in intensity. This means that it can detect and measure differences in the fluorescence or light scattering properties of cells, allowing for the identification and separation of different cell types or subpopulations. By analyzing changes in intensity, flow cytometry can provide valuable information about cellular processes, such as changes in protein expression levels or cellular activation.
2.
Traditional fluorescence microscopy is limited by the following problems:
Correct Answer(s)
A. Difficult to acquire and analyze a statistically large number of events per sample
B. Images are laborious and time consuming to acquire
D. Difficult to quantify images in a standardized and objective manner despite the tremendous amount of information present in each image
Explanation
Traditional fluorescence microscopy is limited by several problems. Firstly, it is difficult to acquire and analyze a statistically large number of events per sample. This means that the data collected may not be representative or reliable. Additionally, the process of acquiring images is laborious and time-consuming, which can hinder efficiency and productivity. Moreover, the resolution for blood cells is poor, leading to limitations in studying and analyzing them. Lastly, despite the vast amount of information present in each image, it is challenging to quantify the images in a standardized and objective manner. This hampers the ability to draw accurate conclusions from the data.
3.
Combining flow cytometry and microscopy in a single platform enables:
Correct Answer(s)
A. Visual feedback for no guesswork gating
B. Discrimination of debris and other artifacts from real cells
C. Identification of cell conjugates or doublets
D. Identification of surface or intracellular staining
E. Quantitative image analysis of rare cells
G. Objective collection and analysis of a statistically relevant number of images per sample
H. High speed image acquisition for statistical microscopy applications
Explanation
Combining flow cytometry and microscopy in a single platform allows for visual feedback, eliminating the need for guesswork in gating. It also enables the discrimination of debris and other artifacts from real cells, ensuring accurate analysis. The platform can identify cell conjugates or doublets, as well as surface or intracellular staining, providing valuable information about cell characteristics. Additionally, it allows for quantitative image analysis of rare cells, enabling researchers to study and understand these cells more effectively. The platform also enables the objective collection and analysis of a statistically relevant number of images per sample, improving the reliability of the results. Lastly, it offers high-speed image acquisition, which is beneficial for statistical microscopy applications.
4.
Primary reason(s) for approaching Microscopists with Amnis technology:
Correct Answer
C. Statistical power ensures that image-based analysis is representative of the sample
Explanation
Approaching Microscopists with Amnis technology is primarily done because of its statistical power, which ensures that the image-based analysis is representative of the sample. This means that the technology is able to provide accurate and reliable results based on the images captured, allowing for a more comprehensive understanding of the sample being analyzed. The other options mentioned, such as superior resolution, pseudocolor representation, and the ability to stitch images together, may also be beneficial features of Amnis technology, but the main reason for approaching Microscopists is the statistical power it offers.
5.
Imaging in flow provides:
Correct Answer(s)
A. A means of verifying gates
B. The ability to measure the spatial distribution of signals
C. The ability to measure changes in appearance and intensity with statistical power
Explanation
Imaging in flow provides a means of verifying gates, which means it allows for the confirmation and validation of the gating strategy used in flow cytometry analysis. It also provides the ability to measure the spatial distribution of signals, allowing researchers to understand the localization and distribution of specific molecules or markers within a sample. Additionally, imaging in flow enables the measurement of changes in appearance and intensity with statistical power, which means it can accurately quantify and analyze variations in signal intensity and morphology.
6.
What is the advantage of having an image for immunophenotyping experiments?
Correct Answer(s)
A. Enables gating with confidence
B. Helps discrimate single cells expressing two surface markers from doublets
C. Verifies overall health of cells
D. Provides visual validation of staining protocol
Explanation
Having an image for immunophenotyping experiments provides several advantages. Firstly, it enables gating with confidence, meaning that researchers can accurately identify and analyze specific cell populations within a sample. Additionally, it helps discriminate single cells expressing two surface markers from doublets, ensuring accurate identification of individual cells. Moreover, it verifies the overall health of cells, allowing researchers to assess the quality of the sample and ensure reliable results. Lastly, having an image provides visual validation of the staining protocol, confirming that the desired markers are being properly labeled and detected.
7.
Can you analyze nuclear translocation of NFkB with a flow cytometer?
Correct Answer
D. No, there is not really a discernible difference in FSC, SSC, or fluorescence intensity
Explanation
The correct answer states that it is not possible to analyze nuclear translocation of NFkB with a flow cytometer because there is not a discernible difference in forward scatter (FSC), side scatter (SSC), or fluorescence intensity. This suggests that the flow cytometer cannot accurately detect the changes in these parameters that would indicate nuclear translocation of NFkB. Therefore, an alternative method should be used to analyze nuclear translocation.
8.
Can you analyze nuclear translocation of NFkB with a manual fluorescence microscope?
Correct Answer
C. Yes, but results can be questionable due to subjective analysis of a small number of cells per sample, especially for rare events
Explanation
The correct answer is "yes, but results can be questionable due to subjective analysis of a small number of cells per sample, especially for rare events." This is because while it is possible to analyze nuclear translocation of NFkB with a manual fluorescence microscope, the results may not be entirely reliable due to the subjective analysis of a small number of cells per sample. This is particularly true for rare events, where the limited number of cells observed may not accurately represent the overall population.
9.
Can you analyze nuclear translocation of NFkB using western blotting?
Correct Answer
B. Yes, but you cannot get single cell translocation status or quantify translocation in subpopulations wihtin the sample
Explanation
The correct answer is yes, but you cannot get single cell translocation status or quantify translocation in subpopulations within the sample. This is because western blotting can provide information about the overall presence of NFkB in the nuclear fraction, but it cannot determine the translocation status of individual cells or quantify the extent of translocation in specific subpopulations within the sample.
10.
Which kind of samples are well-suited for analysis using Amnis cytometers?
Correct Answer(s)
A. Whole blood leukocytes
E. Adherent cells adaptable to flow
F. Non-adherent cell lines
11.
Which of the following assay(s) are well-suited for analysis using Amnis cytometers?
Correct Answer(s)
A. Discrimination of double-positive single cells from conjugate artifacts
B. Quantification of chemokine-induced shape change in human monocytes
C. Quantification of nuclear translocation in rare, immunopHenotypically-defined subsets within whole blood leukocyte preps
D. Evaluation of parasite burden within RBC
Explanation
Amnis cytometers are well-suited for various types of assays. They are particularly useful for discriminating double-positive single cells from conjugate artifacts, quantifying chemokine-induced shape change in human monocytes, quantifying nuclear translocation in rare, immunophenotypically-defined subsets within whole blood leukocyte preps, and evaluating parasite burden within red blood cells (RBC). However, they may not be the best choice for quantifying cirrhosis-induced scarring on liver tissue sections.
12.
What are some advantages of conventional flow cytometry over conventional fluorescence microscopy?
Correct Answer(s)
A. Throughput is higher, allowing greater statistical power
C. Rare event analysis
Explanation
Conventional flow cytometry has a higher throughput compared to conventional fluorescence microscopy, which means it can analyze a larger number of cells or particles in a shorter amount of time. This higher throughput allows for greater statistical power in data analysis, as larger sample sizes can be obtained. Additionally, flow cytometry is particularly advantageous for rare event analysis, as it can efficiently identify and analyze low-frequency events within a sample. This technique also provides quantitative parameters such as intensity, shape, and co-localization metrics, allowing for more comprehensive data analysis.
13.
What are some advantages of FlowSight Basic cytometry over conventional flow cytometry?
Correct Answer(s)
B. More information per probe
E. Image visualization, enabling guess-free gating
Explanation
FlowSight Basic cytometry has several advantages over conventional flow cytometry. Firstly, it provides more information per probe, allowing researchers to gather more detailed data from each sample. Additionally, it offers image visualization, which enables guess-free gating and improves the accuracy of data analysis. This feature is particularly useful in identifying and analyzing rare events. By providing quantitative parameters such as intensity, shape, and co-localization metrics, FlowSight Basic cytometry allows for a more comprehensive analysis of the samples. Overall, these advantages enhance the efficiency and accuracy of cytometry analysis.
14.
What are some advantages of FlowSight Quantitative Imaging cytometry over conventional flow cytometry?
Correct Answer(s)
B. More information per probe
D. Quantitative parameters include intensity, shape and co-localization metrics
E. Image visualization, enabling guess-free gating
Explanation
FlowSight Quantitative Imaging cytometry offers several advantages over conventional flow cytometry. Firstly, it has a higher throughput, which allows for greater statistical power in analyzing larger sample sizes. Additionally, it provides more information per probe, allowing for a more comprehensive analysis of the sample. The quantitative parameters it offers include intensity, shape, and co-localization metrics, providing a more detailed understanding of the cellular characteristics. Moreover, the image visualization feature of FlowSight enables guess-free gating, making the analysis more accurate and reliable. Overall, these advantages make FlowSight Quantitative Imaging cytometry a more advanced and efficient technique compared to conventional flow cytometry.
15.
FlowSight:
Correct Answer(s)
A. Is designed for traditional flow applications
B. Provides imagery in up to 12 channels of every acquired cell
C. Acquires brightfield, darkfield, and fluorescence images
D. Can measure size, SSC and Intensity, plus additional morpHology-based parameters
E. Has a 785 nm Laser dedicated to Side Scatter and optimized for WBCs
F. Is an excellent protocol development tool
H. Can generate FCS-compatible data
Explanation
The correct answer includes all the features and capabilities of FlowSight. It is designed specifically for traditional flow applications and is capable of providing imagery in up to 12 channels for every acquired cell. It can acquire brightfield, darkfield, and fluorescence images. Additionally, it can measure size, SSC and Intensity, along with additional morphology-based parameters. It also has a 785 nm Laser dedicated to Side Scatter and optimized for WBCs. FlowSight is considered an excellent protocol development tool and is able to generate FCS-compatible data.
16.
FlowSight Quantitative Imaging (QI) upgrade includes:
Correct Answer(s)
A. 10 Channel brightfield for flexibility in flow panel design
C. Enhanced image analysis features with IDEAS
D. Enhanced 488 laser power for high sensitivity imaging
E. Wizard-based analysis for ease of use
Explanation
The FlowSight Quantitative Imaging (QI) upgrade includes several features that enhance its functionality. These include a 10 Channel brightfield option, which allows for greater flexibility in designing flow panels. The upgrade also includes enhanced image analysis features with IDEAS, providing more advanced and detailed analysis capabilities. Additionally, the upgrade offers enhanced 488 laser power, which improves sensitivity in imaging. Lastly, the upgrade includes a wizard-based analysis feature, making the software easier to use and navigate.
17.
Autosampler: which of the following is correct?
Correct Answer(s)
B. Can run multiple experiments on the same 96-well plate
C. Can be configured to send email notification for well failures
D. Can be run overnight
Explanation
The given answer is correct because it states that the autosampler can run multiple experiments on the same 96-well plate, which allows for efficient use of resources. Additionally, it mentions that the autosampler can be configured to send email notifications for well failures, which helps in identifying and addressing any issues promptly. Lastly, it states that the autosampler can be run overnight, indicating its ability to operate for extended periods without supervision.
18.
Which system would better discriminate dim signals from background?
Correct Answer
B. B (on right)
Explanation
The system on the right (B) would better discriminate dim signals from the background compared to the system on the left (A). This could be because the system on the right appears to have a larger aperture or lens, which allows more light to enter and improves the signal-to-noise ratio. Additionally, the system on the right seems to have a higher sensitivity or gain, as indicated by the larger output signal. These factors contribute to better discrimination of dim signals from the background.
19.
The Experiment Evaluation Worksheet:
Correct Answer(s)
A. Directs researcher toward applications that leverage unique benefits of the platform
B. Clarifies goals of the sample evaluation
C. Provides specific details for planning the evaluation samples
E. Is used to design a practical experiment that efficiently evaluates the platform’s unique value
F. Begins documentation of the evaluation process
Explanation
The Experiment Evaluation Worksheet serves multiple purposes. It directs the researcher towards applications that leverage the unique benefits of the platform, clarifies the goals of the sample evaluation, and provides specific details for planning the evaluation samples. Additionally, it is used to design a practical experiment that efficiently evaluates the platform's unique value and begins the documentation of the evaluation process.
20.
Dot plots and histograms in IDEAS:
Correct Answer(s)
B. Are also used in standard flow cytometry
C. Can be used to plot many types of parameters in addition to fluorescence intensity
D. Are linked directly to the images they represent, enabling visual verification of dots and histogram bins
Explanation
Dot plots and histograms in IDEAS are not limited to recapitulating plots from conventional flow cytometers. They are also utilized in standard flow cytometry. Additionally, they have the capability to plot various types of parameters apart from fluorescence intensity. Moreover, dot plots and histograms in IDEAS are directly connected to the images they represent, allowing for visual verification of dots and histogram bins.
21.
Which of the following statement(s) regarding the ImageStream and FlowSight are true?
Correct Answer(s)
B. All quantitave data for both platforms are derived from images
C. The Quantitative Imaging (QI) capability comes standard on the ISX, but is an option for FlowSight
E. Both enable image visualization and therefore guess-free gating
Explanation
The ImageStream and FlowSight both provide 40X imagery. The statement that all quantitative data for both platforms are derived from images is true. The Quantitative Imaging (QI) capability comes standard on the ISX, but is an option for FlowSight. Both platforms can be upgraded with the Extended Depth of Field (EDF) element. Additionally, both platforms enable image visualization and therefore guess-free gating.
22.
Consider the ImageStream endosomal and lysosomal co-localization experiment described in the Jan 5 Introductory webinar. Which of the following statement(s) are true?
Correct Answer(s)
A. ImageStream provides objective collection of images, thereby eliminating operator collection bias that can occur with manual image acquisition on a traditional fluorescence microscope
B. ImageStream high acquisition rate provides a statistically significant number of events to analyze, allowing the researcher to conclude that there is a time-dependent accumulation of CpGB in the lysosome
Explanation
The ImageStream provides objective collection of images, eliminating operator collection bias that can occur with manual image acquisition on a traditional fluorescence microscope. Additionally, the high acquisition rate of the ImageStream provides a statistically significant number of events to analyze, allowing the researcher to conclude that there is a time-dependent accumulation of CpGB in the lysosome.
23.
What are the primary value propositions of Amnis technology?
Correct Answer(s)
A. The value that imagery brings to flow cytometry
B. The value that population statistics brings to microscopy
C. Visual feedback for no guesswork gating
D. The power of combining flow cytometry with microscopy as a protocol development tool
Explanation
The primary value propositions of Amnis technology include the value that imagery brings to flow cytometry, the value that population statistics brings to microscopy, visual feedback for no guesswork gating, and the power of combining flow cytometry with microscopy as a protocol development tool. These value propositions highlight the benefits and advantages of using Amnis technology in both flow cytometry and microscopy applications, such as improved imaging capabilities, enhanced statistical analysis, more accurate gating, and the ability to combine different techniques for protocol development.