Polychromatic flow cytometry in evaluating rheumatic disease patients

Table 2 Composition of the staining panels for human B-cell phenotyping

	Memory panel		Transitional panel		Plasma cell panel
Fluorochrome	Specificity	Clone	Specificity	Clone	Specificity	Clone
FITC	IgD	IA6-2	MTG	Free dye	Ki-67	B56
PE	CXCR3	1C6	IgD		CXCR3	1C6
PE-Alexa610	CD24	SN3	CD24		CD24
PE-Cy5	CD21	B-ly4	IgM	G20-127	CXCR4	12G5
PerCP-Cy5.5	CD38	HIT2	CD38		CD3
PE-Cy7	CD45/B220	RA3/6B2	CD23	EBVCS2	CD38
Pacific Blue	CD3	SP34-2	CD3		VH4-34
Pacific Orange	Live/Dead	Aqua	Live/Dead		Live/Dead
Qdot605	CD27	CLB-27/1	CD27		CD27
APC	CD95	DX2	CD10	HI10a	CD138	B-B4
Alexa680	VH4-34	9G4	VH4-34		IgD
APC-Cy7	CD19	SJ25C1	CD19		CD19

The seven anchor markers and VH4-34 encoded heavy chain are in bold font. Information on the detecting antibody clones for these eight molecules is omitted from the transitional and plasma cell panels, as they are the same as those indicated under the memory panel regardless of the fluorochrome conjugates. For detection in the Red B channel, antibodies used are biotinylated and detected by streptavidin-Alexa680. Note that in the plasma cell panel, both Ki-67 and 9G4 are intracellular staining. APC, allophycocyanin; Aqua, LIVE/DEAD fixable aqua dead cell stain; Cy, cyanine; FITC, fluorescein isothiocyanate; PE, R-phycoerythrin; PerCP, peridinin-chlorophyll protein; Qdot, quantum dot.

ISSN: 1478-6362