SCYM (ASCP) Review: Section 4: Data
ACTUAL UPDATED QUESTIONS AND
CORRECT ANSWERS
M.E.S.F. - CORRECT ANSWERS✅✅Molecules of equivalent soluble fluorochrome
TIFF - CORRECT ANSWERS✅✅Tagged Image File Format: a large file that contains
everything. Other files reduce file size by eliminating data.
My Flowcyt Standard - CORRECT ANSWERS✅✅An attempt to more completely explain
the details of a flow experiment. 12 years old.
Flow Storage Requirements - CORRECT ANSWERS✅✅Never edit raw data. Make a copy
and store data on RAID array system and make a backup copy.
Signal Processing - How a photon becomes data - CORRECT ANSWERS✅✅1. Photons are
converted to photocurrent
2. Photocurrent is amplified, sampled, and digitized
3. Digitized values are binned and stored in list mode file
Low level signals are negatively impacted by - CORRECT ANSWERS✅✅- dark current -
spontaneous signal
- ramen light scatter
- fluorescence of sheath fluid
Baseline Restoration Circuitry - CORRECT ANSWERS✅✅Removes issues from a pulse.
Subtracts constant background during current to pulse conversion. Works like compensation.
Background Correction - CORRECT ANSWERS✅✅Measure of background with no cells
present. Instrument background is subtracted from the positive signals. Introduces statistical
variation.
, Histogram - CORRECT ANSWERS✅✅Limited usefulness. Hard to identify rare events.
Bivariant Plots - CORRECT ANSWERS✅✅Visualization of 2 stains. Density plots provide
context of relative number of events. Be careful of contour plots, as they don't display well.
Always indicate the number of events when using a contour plot.
Higher Dimensional Analysis - CORRECT ANSWERS✅✅Cluster Analysis & 3D plots
Regions - CORRECT ANSWERS✅✅Numerical or geological boundary used to define
characteristics of particles to include for further analysis. drawing regions is subjective,
unless proper controls are used.
Critical Controls - CORRECT ANSWERS✅✅- FMO
- Biological Controls
- Stimulated Samples
- Reference Controls
Controls critical to make region drawing objective - CORRECT ANSWERS✅✅- Unstained
- Assed autofluorescence in each channel
- FMO - helpful for region drawing in multicolor experiments
- Unstimulated Controls - Address background
- Isotype controls - DO NOT USE for region placement
Cutoff %s - CORRECT ANSWERS✅✅Statistical placement of regions
Specialty Gates - CORRECT ANSWERS✅✅- Back gating
- Time gating
- Singlet gating
Hierarchical Gating - CORRECT ANSWERS✅✅Parent/child gates- child has all
characteristics of parent gate
ACTUAL UPDATED QUESTIONS AND
CORRECT ANSWERS
M.E.S.F. - CORRECT ANSWERS✅✅Molecules of equivalent soluble fluorochrome
TIFF - CORRECT ANSWERS✅✅Tagged Image File Format: a large file that contains
everything. Other files reduce file size by eliminating data.
My Flowcyt Standard - CORRECT ANSWERS✅✅An attempt to more completely explain
the details of a flow experiment. 12 years old.
Flow Storage Requirements - CORRECT ANSWERS✅✅Never edit raw data. Make a copy
and store data on RAID array system and make a backup copy.
Signal Processing - How a photon becomes data - CORRECT ANSWERS✅✅1. Photons are
converted to photocurrent
2. Photocurrent is amplified, sampled, and digitized
3. Digitized values are binned and stored in list mode file
Low level signals are negatively impacted by - CORRECT ANSWERS✅✅- dark current -
spontaneous signal
- ramen light scatter
- fluorescence of sheath fluid
Baseline Restoration Circuitry - CORRECT ANSWERS✅✅Removes issues from a pulse.
Subtracts constant background during current to pulse conversion. Works like compensation.
Background Correction - CORRECT ANSWERS✅✅Measure of background with no cells
present. Instrument background is subtracted from the positive signals. Introduces statistical
variation.
, Histogram - CORRECT ANSWERS✅✅Limited usefulness. Hard to identify rare events.
Bivariant Plots - CORRECT ANSWERS✅✅Visualization of 2 stains. Density plots provide
context of relative number of events. Be careful of contour plots, as they don't display well.
Always indicate the number of events when using a contour plot.
Higher Dimensional Analysis - CORRECT ANSWERS✅✅Cluster Analysis & 3D plots
Regions - CORRECT ANSWERS✅✅Numerical or geological boundary used to define
characteristics of particles to include for further analysis. drawing regions is subjective,
unless proper controls are used.
Critical Controls - CORRECT ANSWERS✅✅- FMO
- Biological Controls
- Stimulated Samples
- Reference Controls
Controls critical to make region drawing objective - CORRECT ANSWERS✅✅- Unstained
- Assed autofluorescence in each channel
- FMO - helpful for region drawing in multicolor experiments
- Unstimulated Controls - Address background
- Isotype controls - DO NOT USE for region placement
Cutoff %s - CORRECT ANSWERS✅✅Statistical placement of regions
Specialty Gates - CORRECT ANSWERS✅✅- Back gating
- Time gating
- Singlet gating
Hierarchical Gating - CORRECT ANSWERS✅✅Parent/child gates- child has all
characteristics of parent gate
