Reduction of interlaboratory variability in flow cytometric immunophenotyping by standardization of instrument set‐up and calibration, and standard list mode data analysis

Use of internal controlT‐cell populations in the flow cytometric evaluation forT‐cell neoplasms

“…B ) . The need for standardization of flow cytometric assays between instruments is well recognized and has led many laboratories to switch from using target voltages to using application‐specific target channels . The goal of this strategy is to have non‐neoplastic populations consistently fall in the same position on dot‐plots, regardless of the instrument used.…”

Apheresis‐induced platelet activation:comparison of three types of cell separators

“…The samples were analyzed within 12 hours in a flow cytometer (FACSort) equipped with a data management system (FACStation) and software (CellQuest) (all: Becton Dickinson). A common window of analysis 27 was set with microbead reference standards (QC Windows, Flow Cytometry Standards Corp., San Juan, PR). Calibration allowing conversion of the fluorescence intensity into molecules of equivalent soluble fluorochrome (MESF) units 27 was performed weekly with microbeads (Quantum 26 and Quantum 27, Flow Cytometry Standards).…”

“…A common window of analysis 27 was set with microbead reference standards (QC Windows, Flow Cytometry Standards Corp., San Juan, PR). Calibration allowing conversion of the fluorescence intensity into molecules of equivalent soluble fluorochrome (MESF) units 27 was performed weekly with microbeads (Quantum 26 and Quantum 27, Flow Cytometry Standards). The fluorescence intensity was controlled daily with fluorescent microspheres (Standard‐Brite Coulter Corp., Miami, FL) according to the manufacturer's instructions.…”

Analysis of variation in results of CD34+ hematopoietic progenitor cell enumeration in a multicenter study

“…In addition, centrally prepared suspensions of mononuclear cells from a healthy donor that were unstained or had been stained with CD4/FITC, CD8/PE, or LDS-751 were supplied for adjustment of electronic compensation for spectral overlap and adjustment of FSC photodiode and SSC photomultiplier settings. Instruments had to be set up as previously described (8). In brief, standardized positioning in ''sample space'' (i.e., the characteristics of the sample and their relationships, which are independent of the instrument) of the instrument's ''window of analysis'' (i.e., the portion of sample space analyzed by the flow cytometer) had to be achieved for each parameter by placement of the respective FL signals of CD4 1 lymphocytes (FSC and SSC parameters) or QC3 microbeads (all FL parameters) into predefined target channels.…”