Sample preparation solutions for Flow Cytometry
Red blood cells (RBC) are the major component of blood as well as abundant in blood-rich organs. Presence of RBC in a sample is not desirable when analyzing leukocyte populations by flow cytometry and therefore efficient lysis of erythrocytes from blood samples must performed prior to analysis.
We offer you the essential portfolio of lysing, fixative and permeabilizating reagents for standardized laboratory workflows.
Formaldehyde-based RBC lysing solutions
Formaldehyde-based agents are designed to lyse erythrocytes under mild hypotonic conditions while the leukocytes are spared from destruction by mild simultaneous fixing that renders stained populations stable and thus allows enough time for flow cytometry measurements, including next day sample acquisition or repeat analysis if needed.
EXCELLYSE Easy is a one step reagent that is intended to prepare a blood sample for flow cytometry analysis. The reagent is well compatible with various blood sample staining approaches and techniques, e.g. Stain-Lyse-Wash or Stain-Lyse-No Wash protocols. Use of the reagent is Easy: just add appropriate volume of diluted (1X) solution to antibody-stained sample. The simplicity of the sample preparation makes the reagent an excellent choice for routine and standardized immunophenotyping in clinical laboratories. EXCELLYSE Easy is supplied in a 10X concentrated form that ensures the stability of the fixative in the solution and hence the long shelf life (36 months). Product must be diluted with deionized water prior to use. Diluted (1X) solution still maintain the substantial in-use stability at room temperature (1 month). Therefore, it may be convenient to prepare 1X solution in HDPE or glass containers with top dispensers for daily laboratory use.
EXCELLYSE I is a two-step reagent that is intended to prepare a blood sample flow cytometry analysis. The reagent is well compatible with various blood sample staining approaches and techniques, e.g. Stain-Lyse-Wash or Stain-Lyse-No Wash protocols. The major advantage of EXCELLYSE I is that reagent is supplied in a ready-to use format. However, since the reagent is a concentrated fixative solution, it requires addition of deionized water in the sample tube to create hypotonic conditions for efficient RBC lysis. Procedure leads to a significant erythrocyte lysis, fixation of leukocytes and antigen preserving with well distinguished and homogenous leukocyte populations without considerable light scatter changes. Content of formaldehyde and methanol disrupts the integrity of the cellular membranes and partial permeabilization, however, in order to be used in the intracellular staining procedures, EXCELLYSE I has to be complemented with a suitable detergents and non-specific background blocking agents to achieve adequate fluorescence signals for the target antigens.
Fixative-free RBC lysing solution
EXCELLYSE Live is isotonic ammonium chloride based RBCs lysing solution that does not permeabilize leukocyte cell membranes. The solution does not contain any fixative agent, so leukocytes remain viable after erythrocyte lysis. EXCELLYSE Live is supplied as 10X concentrated solution with proprietary stabilizing agents that provide significantly increased shell life stability (36 months) if compared to the standard ACK (Ammonium-Chloride-Potassium) lysis buffer. Product must be diluted with deionized water and freshly prepared prior to use. Diluted (1X) solution can be conveniently used in for whole blood or bone marrow aspirate sample processing procedures when the rare cell subpopulations are to be detected (so called bulk lysis protocol). EXCELLYSE Live is also incorporated in the assay of human CD34+ progenitor cells using CD34 QuantiFlowEx Kit.
Cell Fixation and Permeabilization Kit
For simultaneous flow cytometry analysis of intracellular and cell-surface antigens in the same cell we can provide you with EXCELLYSE XPerm, a set of two buffered solutions that were carefully selected to preserve the surface stained fluorescence signals and the morphological light scatters of the cells, while reducing the background fluorescence due to the non-specific binding of antibodies used for intracellular staining.
EXCELLYSE XPerm consist of fixation reagent (Solution A) and permeabilization reagent (Solution B) and it is suitable for the analysis of the most common clinically relevant intracellular markers.
View examples of experimental data here.