M.E.S.F. - ANSWER Molecules of equivalent soluble fluorochrome
My Flowcyt Standard - ANSWER An attempt to more completely explain the details of a
flow experiment. 12 years old.
TIFF - ANSWER Tagged Image File Format: a large file that contains everything. Other files
reduce file size by eliminating data.
Flow Storage Requirements - ANSWER 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 - ANSWER 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 - ANSWER - dark current - spontaneous sig-
nal
- ramen light scatter
- fluorescence of sheath fluid
Baseline Restoration Circuitry - ANSWER Removes issues from a pulse. Subtracts constant
background during current to pulse conversion. Works like compensation.
1
, Background Correction - ANSWER Measure of background with no cells present. Instru-
ment background is subtracted from the positive signals. Introduces statistical variation.
Histogram - ANSWER Limited usefulness. Hard to identify rare events.
Bivariant Plots - ANSWER Visualization of 2 stains. Density plots provide context of rela-
tive 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 - ANSWER Cluster Analysis & 3D plots
Regions - ANSWER 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 - ANSWER - FMO
- Biological Controls
- Stimulated Samples
- Reference Controls
Controls critical to make region drawing objective - ANSWER - Unstained - Assed auto-
fluorescence 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 - ANSWER Statistical placement of regions
Specialty Gates - ANSWER - Back gating
2
My Flowcyt Standard - ANSWER An attempt to more completely explain the details of a
flow experiment. 12 years old.
TIFF - ANSWER Tagged Image File Format: a large file that contains everything. Other files
reduce file size by eliminating data.
Flow Storage Requirements - ANSWER 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 - ANSWER 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 - ANSWER - dark current - spontaneous sig-
nal
- ramen light scatter
- fluorescence of sheath fluid
Baseline Restoration Circuitry - ANSWER Removes issues from a pulse. Subtracts constant
background during current to pulse conversion. Works like compensation.
1
, Background Correction - ANSWER Measure of background with no cells present. Instru-
ment background is subtracted from the positive signals. Introduces statistical variation.
Histogram - ANSWER Limited usefulness. Hard to identify rare events.
Bivariant Plots - ANSWER Visualization of 2 stains. Density plots provide context of rela-
tive 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 - ANSWER Cluster Analysis & 3D plots
Regions - ANSWER 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 - ANSWER - FMO
- Biological Controls
- Stimulated Samples
- Reference Controls
Controls critical to make region drawing objective - ANSWER - Unstained - Assed auto-
fluorescence 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 - ANSWER Statistical placement of regions
Specialty Gates - ANSWER - Back gating
2
