SCYM (ASCP) Flow cytometry EXAM STUDY GUIDE 2025/2026 COMPLETE QUESTIONS BANK WITH VERIFIED CORRECT ANSWERS|| 100% GUARANTEED PASS <RECENT VERSION>

SCYM (ASCP) Flow cytometry EXAM STUDY GUIDE 2025/2026 COMPLETE QUESTIONS BANK WITH VERIFIED CORRECT ANSWERS|| 100% GUARANTEED PASS &lt;RECENT VERSION&gt; 1. SCYM(ASCP) - ANSWER Specialist in Cytometry certification. 2. SCYM(ASCPi) - ANSWER International Specialist in Cytometry certification. 3. Examination Duration - ANSWER The SCYM(ASCP) and SCYM(ASCPi) certification examination is composed of 100 questions given in a 2 hour 30 minute time frame. 4. Examination Question Types - ANSWER Examination questions may be both theoretical and/or procedural. 5. Theoretical Questions - ANSWER Measure skills necessary to apply knowledge, calculate results, and correlate patient results to disease states. 6. Procedural Questions - ANSWER Measure skills necessary to perform laboratory techniques and follow quality assurance protocols. 7. Regulatory Questions - ANSWER Based on U.S. sources (e.g., AABB, FDA, CLIA, etc.). 8. Instrumentation Content Area - ANSWER Principles of fluidic, optical, and electronic instrumentation including troubleshooting (15 - 20%). 9. Panel/Experiment Design Content Area - ANSWER Sample source, sample integrity, sample preparation and staining, cell enrichment, and assay development (25 - 30%). 10. Applications Content Area - ANSWER Includes immunophenotyping, functional assays, multiplex bead assays, solid organ transplant, stem cell analysis, cell cycle / DNA ploidy, rare event analysis, small particle analysis, fetal hemoglobin assay, cell sorting, imaging cytometry, and mass cytometry (25 - 30%). 11. Data Content Area - ANSWER Data standards, signal processing, data display, gating, statistical methods, common data modeling techniques, quantitative cytometry, and troubleshooting (15 - 20%). 12. Laboratory Operations Content Area - ANSWER Quality control, assay validation, safety, and laboratory administration (10 - 15%). 13. Fluidics - ANSWER Hydrodynamic focusing and properties of sheath fluids. 14. Sample Delivery Methods - ANSWER Includes syringe pump, pressure based, vacuum, acoustic. 15. Signal Processing - ANSWER Techniques used to enhance and analyze signals, including binning and compensation. 16. Statistical Methods - ANSWER Analytical techniques for summarizing data, such as central tendency and standard deviation. 17. Quantitative Cytometry - ANSWER Measurement techniques that provide quantitative data, such as molecules of equivalent soluble fluorochrome. 18. Biosafety procedures - ANSWER Protocols to ensure safety when handling biological materials, including PPE and decontamination. 19. Instrument quality control - ANSWER Methods to ensure that laboratory instruments are functioning correctly, including calibration. Hydrodynamic Focusing - ANSWER Most modern flow cytometers tightly position the sample for optical analysis via hydrodynamic focusing. Here, a carrier fluid called the sheath fluid is used to position the sample of cells into a single file for optical interrogation. 20. Hydordynamic focusing and sheath fluids - ANSWER The central stream (sample stream) is focused and surrounded by the secondary slower stream (sheath fluid). The shape and size of the flow cell is crucial to hydrodynamic focusing, and traditionally the cell is nozzle shaped. ... In a flow cytometer, the sheath fluid pressure is constant while the sample fluid is adjusted 21. Sample Pressure and the Sheath Pressure - ANSWER The difference between the sample pressure and the sheath pressure is the differential pressure. This controls the width of the core stream and the total number of cells passing the laser intercept. 22. Differential pressure based flow cytometers - ANSWER Differential pressure based flow cytometers currently dominate the market. These systems have two pressure regulators. The first is at a constant pressure that sets how fast the fluids runs at. The second is regulated by the investigator (like on this LSR-II control panel). 23. Generation of differential pressure (syringe pump, pressure based) - ANSWER Low differential pressure allows the cells to move past the interrogation point one at a time. .... One kind involves generating pressure using a pump and regulator system ... Differential pressure based fluidic system. ... peristaltic and/or syringe pumps to deliver the sample into the instrument. 24. Stability - ANSWER Specimen stability assessment is critical in all cases where samples will not be assayed within a few hours of collection. Certain markers or cellular subsets may be lost or altered during the storage/shipment of the sample (typically whole blood or bone marrow). Antigen expression, cellular composition, and viability can change over time in the anti‐coagulant tube. The time points for the stability evaluation should be based on when the samples are expected to arrive at the testing laboratory, and should include at least one time point beyond the expected transit time. The generally accepted change for a particular marker from the baseline specimen value and the stored specimen value is 20% difference or a change within the acceptable assay precision (10% to 30% CV). 25. Carryover - ANSWER It is critical to evaluate instrument carryover from one sample tube to the other when evaluating high‐sensitivity assays reporting rare events. Carryover can be assessed during the initial validation by placing a tube with buffer between sample tubes. Data from the blank tubes would be evaluated in the same gating template as the samples. 26. Reference Ranges - ANSWER Reference ranges (age, gender, disease specific ranges, healthy ranges) are essential in the interpretation of clinical chemistry laboratory results but are not always required for flow cytometric methods. Procedure for establishing reference ranges can be found in the Clinical and Laboratory Standards Institute (CLSI) guideline EP28‐A3c 27. Documentation - ANSWER Validation should follow a three‐step approach: 1. Say It (the Validation Plan or Protocol) 2. Do It (the Experimental Phase) 3. Prove It (the Validation Report) 28. Documentation is critical because in a regulated environment, "if you don't document what you did, it didn't happen." 29. Method Validation Plan - ANSWER The Method Validation Plan or Validation Protocol provides detailed documentation of the validation experimental design. The Validation Plan should include: 1. A full description of the method including gating strategy and list of the assay read‐outs to be validated. The source (disease, healthy) and type of validation samples, including anticoagulants when using whole blood or bone marrow samples. 2. Quality control material, if applicable. The control material should mimic the actual sample as closely as possible. 3. Critical reagents (manufacturer, catalog number): For monoclonal antibodies, the fluorochrome and clone designation must be specified. 30. Method Validation Report - ANSWER Method validation report describes the results of the experimental phase. Validation Report should mirror the Validation Plan. Any deviations from the Validation Plan which occurred during the experimental phase should be explained clearly. The validation report should include: 1. A reagent table with the lot number and expiration dates of all reagents. 2. The statistical results clearly summarized with table summaries and/or figures: a- If any of the parameters did not meet the acceptance criteria, it should be discussed within that section of the document. b- if any outliers were identified, then exclude them from the statistical analysis—justification must be provided along with the statistical tool used to identify outliers. Note that the outliers must be included in the report. 3. Individual results should be presented in an appendix in table format. Changes in the gating strategy or other deviations from the validation plan must be clearly described in the validation report. The documentation for deviations typically includes an impact assessment but must follow the laboratory's institution's quality processes. 4. Copies of the gated data (from the data analysis software) for every sample should be available in a validation binder and/or electronic format. In addition, the location of the list mode files should be either included in the Validation Report or described in an SOP. 31. Standard operating procedure (SOP) - ANSWER Information included in the SOP: The reagent tables including storage and handling instructions. Complete details for sample accessioning, processing, and reporting. Detailed instrument setup and compensation procedure. Detailed instrument acquisition procedure (e.g., how many events to collect). Detailed gating instructions. Additional specific requirements for a method SOP depend on the regulatory environment in which the testing is conducted. 32. Limited Assay Validation - ANSWER The Limited Assay Validation represents the minimal recommended parameters for research environments and non‐regulated laboratories. In this example, it is assumed that the samples will not be shipped to a testing facility and assumes one operator and one instrument; thus, specimen stability as well as inter‐operator and inter‐instrument variation are not included in this validation plan. Note that although sample stability is not included in this validation plan, it is assumed that stability was demonstrated during assay optimization and that samples are analyzed within that window. 33. Initial Assay Validation - ANSWER Initial Assay Validation is performed for a new assay in a laboratory. Using the fit‐for‐purpose validation approach, the validation parameters should be selected based on the intended use of the assay. The example shown below represents the minimal validation protocol for low and moderate risk assays in regulated laboratories. 34. CD34+ Absolute Counts - ANSWER Direct measurement of CD34+ blood stem cell absolute counts by flow cytometry. ... Whole blood was stained with a phycoerythrin-conjugated anti-CD34 monoclonal antibody, and, after the lysis of red blood cells, CD34+ cells were counted in a fraction of the lymphocyte and monocyte gate. 35. What is Flow Cytometry - ANSWER Flow cytometry is a technology that facilitates multiparametric analysis at the single‐cell level and is widely used in research settings, diagnostic laboratories, and drug development. It is also used in a variety of other settings, such as marine biology and food science. In order to ensure that the data generated are reliable, every laboratory should perform some level of analytical method validation. This protocol provides a roadmap for validation which can be applied in any type of laboratory setting and will answer the questions, "what is method validation?" and "why should I care about it?" 36. Solid Organ Transplant (HLA Crossmatch) - ANSWER This test is used primarily for solid organ transplant candidates to assess the suitability of a potential donor. Positive cross matches detected by flow cytometry suggest the presence of preformed anti-donor HLA antibodies and carry a risk of accelerated rejection and damage to the graft. These sensitive cross matches are interpreted in terms of the patient's sensitization history and the quality of the donor organ. 37. Immunophenoyping - ANSWER Immunophenotyping using flow cytometry has become the method of choice in identifying and sorting cells within complex populations, for example the analysis of immune cells in a blood sample. 38. Calcium Flux - ANSWER The principle of flow cytometric Ca2+ flux measurement is based on changes in fluorescence intensity or emission wavelength of a fluorophore following chelating of calcium ions 39. Granulomatous Disease - ANSWER A diagnosis of chronic granulomatous disease is often suspected based on the presence of characteristic signs and symptoms. Specialized blood tests, such as the nitroblue tetrazolium test and/or flow cytometry with dihydrorhodamine, can then be ordered to confirm the diagnosis. 40. PhosphoFlow - ANSWER Proteins can be phosphorylated at 3 residues: Serine, Threonine, or Tyrosine. Phosphoprotein antibodies must be able to resolve a single change at the appropriate site in the presence of non phosphorylated targets. When screening hybridomas, manufacturers must screen for both positive binding and absence of binding to the non phosphorylated target. 41. Cytokine Detection - ANSWER Flow cytometry (FCM) is a highly effective technique that detects intracellular cytokines using specific fluorescence-labeled antibodies. The common steps of this assay include cell collection, fixation, permeabilization, blocking, intracellular staining and analysis by FCM. 42. CD3 Marker - ANSWER T cells (Mature T Cells) 43. CD4 Marker - ANSWER T helper (Th), dimly expressed monocles, used for immunophenotyping such as HIV (CD4 Counts) 44. CD8 Maker - ANSWER Cytotoxic TCells and Marcorphages 45. CD45 Marker - ANSWER leukocytes, leukemic cells are dim 46. CD19 Marker - ANSWER B Cells