inverted CommasI can’t recommend FloCyte enough to people. I wish everyone who prepares flow samples – even if they don’t run them – would take FloCyte’s courses



Certification Exam Preparation Course

FloCyte offered the first preparation course for the new ICCS and ISAC certification examination!    Qualification to take this test will be the same as those to take the exam:   Applicants must have a Bachelors degree or equivalent and two years of cytometry work experience, or four years of cytometry work experience.

Syllabus of the course also follows the outline of topics covered on the exam in approximately the same proportion as the exam.   Order of the topics as they are presented may differ.

I. Instrumentation   20%

Fluidics 5%

  •      Hydrodynamic focusing and properties of sheath fluids
  •      Generation of differential pressures (e.g., syringe pump, pressure based)

Optics 9%

  •         Optical filters (e.g., long pass, band pass, short pass, dichroics, neutral density, polarizing)
  •         Light source (e.g., laser type, laser line, arc lamp, led)
  •         Lenses (e.g., beam shape, collecting, focusing, objective)
  •          Optical pathway (e.g., transmission, reflection, interrogation point, collinear, spatial separation, light scatter)

Electronics 6%

  •          Amplifiers (e.g., Linear, Logarithmic)
  •          Detectors (e.g., photomultiplier tube, photodiode, CCD camera, avalanche photon detector)
  •          Digital vs. analog systemsNoise
  •          Pulse measurement (e.g., time delay, window extension, area, width, Coulter impedance)
  •          Threshold/discriminator


II. Sample 10%

  •         Sample source ( e.g., beads, microspheres, solid tissue, body fluids, subcellular components, cultured cells, microorganisms, plants, whole organisms)
  •         Sample integrity (e.g., collection, handling, storage viability)
  •         Sample preparation and staining (e.g., disaggregation, lysing agents, aggregates, filtering, fixation, permeabilization)
  •         Cell enrichment (e.g., cell sorting, density gradient isolation, magnetic beads)

III. Data 19%

  •         Data standards (e.g., image file format, FCS format, metadata, multichannel data, XML, storage requirements)
  •         Signal processing (e.g., binning, compensation, pulse processing, baseline restoration, background  correction)
  •         Data display (e.g., types of displays, transformations)
  •         Gating (e.g., hierarchical vs. Boolean gating, gates, regions)
  •         Statistical methods (e.g., central tendency, standard deviation, CV, KS statistics, cluster analysis, principal component analysis, discriminant analysis)
  •         Common data modeling techniques (e.g., DNA ploidy, cell cycle analysis, proliferation, phenotyping, ratiometric)
  •         Quantitative cytometry (e.g., molecules of equivalent soluble fluorochrome[MESF], absolute counts)

IV. Safety 3%

  •         Biosafety procedures (e.g., biosafety categories, Personal Protective
  •         Equipment, specimen transport and preparation precautions, aerosols, decontamination)
  •         Instrument safety (e.g., lasers, electronics)
  •         Chemical safety (e.g., mutagenic agents, cytotoxic agents)
  •         Environmental safety (e.g., waste disposal)

V. Quality Control 10%

  •         Instrument quality control (e.g., optical alignment, detector calibration)
  •         Reagent quality control (e.g., panel verification, titration, lot to lot variation, storage, handling)
  •         Sample integrity
  •         Appropriate sample quality controls selection (internal, external)
  •         Trend analysis and interpretation

VI. Experimental Design 19%

Assay Development 6%

  •         Sample state (e.g., activated, resting, proliferating)
  •         Target (e.g., cell type, subcellular location, molecule)
  •         Assay interpretation (e.g., isotype control, autofluorescence, biological
  •         systems control, background measurement controls)
  •         Assay optimization (e.g., appropriate use of limited sample, frequency of
  •         target, cell seeding, kinetics, scalability, blocking, signal to noise, statistical
  •         design, Z factor)

Reagent Selection 10%

  •         Fluorochrome issues (e.g., antigen density, protein coexpression, optimal
  •         combination, photostability, F/P ratio, spectral overlap and compensation,
  •         quenching)
  •         Probe types (e.g., antibodies, viability/DNA dyes, physiological, tracking,
  •         fluorescent proteins)
  •         Solutions (e.g., buffers, fixatives, chelators, permeabilizing agents)

Assay Validation 3%

  •         Method validation (e.g., accuracy, reproducibility/precision, sensitivity,
  •         specificity, linearity, reference range, robustness)
  •         Method calibration (e.g., standards, controls)

VII. Theoretical Principles 19%

Physical Principles 13%

  •         Properties of light (e.g., refraction, diffraction, polarization, scatter)
  •         Fluorescence (e.g., Stokes shift, excitation and emission, energy transfer, environmental sensitivity [such as pH, polarity, calcium])
  •         Optics (e.g., optical filters, image formation, focal plane, numerical       aperture)
  •         Electronics (e.g., signal detection, amplification, processing)
  •         Fluid dynamics (e.g., laminar flow, stream width, turbulence)
  •         Cell sorting (e.g., Jet‐in‐air, cuvette, droplet formation, drop delay, drop        deflection, fanning, charging)

Biological Principles 6%

  •         Antigen/antibody interaction and antibody structure
  •         Fluorescent proteins structure and properties
  •         Optical properties of cells