B59380–AD
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Specifications
VersaLyse Lysing Solution Ready-for-use
Formulation
Liquid
Volume
100 mL
Number of vials
1 vial
Volume per test
1 mL (ready-for-use when used with no concomitant fixation)
VersaLyse
Lysing Solution
.
1 mL / test, A09777
100 tests; 100 mL
For
In Vitro
Diagnostic Use
INTENDED USE
This reagent is intended for the lysis of red blood cells in the preparation of biological samples for flow cytometry
analysis. VersaLyse is compatible with all types of cytometers and is used with or without washing after incubation,
as long as properly calibrated fluorescent antibodies are used in the protocol. VersaLyse also makes it possible to
treat samples even when they have been washed earlier, in order to eliminate serum immunoglobulins for example
(1,2,3,4,5).
PRINCIPLE
The sample containing the red blood cells to be lysed is exposed to the VersaLyse reagent for at least 10 minutes.
The major active ingredient of VersaLyse is a cyclic amine which, in contact with carbonic anhydrase present in
red blood cells, is transformed into a compound which is highly lytic for these cells.
This reagent is used with specific conjugated antibodies, able to bind to leucocytes due to the antigenic
determinants they express. The specific staining of leucocytes is achieved by incubating the sample with the
antibody or antibodies. The red cells are then removed by lysis and the leucocytes, unaffected by this process,
are analyzed by flow cytometry.
Preparations must be stored between 2 and 8°C and analyzed rapidly using flow cytometry. In the case of a delay
of several hours before analysis, they may be fixed with PBS containing 0.1% formaldehyde (see the technical
leaflet of the IOTest 3 Fixative Solution – Ref. A07800 – to prepare this PBS with 0.1% formaldehyde). In this
case, prior washing is necessary.
The flow cytometer measures light diffusion and the fluorescence of cells. It makes possible the delimitation of the
population of interest within the electronic window defined on a histogram, which correlates the orthogonal diffusion
of light (Side Scatter or SS) and the diffusion of narrow-angle light (Forward Scatter or FS). Other histograms
combining two of the different parameters available on the cytometer can be used as an aid in the gating stage
depending on the application chosen by the user.
The fluorescence of the delimited cells is analyzed in order to distinguish the positively-stained events from the
unstained ones. The results are expressed as a percentage of positive events in relation to all the events acquired
by the gating.
WARNING AND PRECAUTIONS
1. Do not use the reagent beyond the expiry date.
2. Do not store in the refrigerator, do not freeze.
3. Minimize exposure to light during incubation with fluorescent antibodies.
4. Avoid microbial contamination of the reagents, or false results may occur.
5. Formaldehyde is toxic and allergenic. It is thought to be carcinogenic. Never pipette by mouth and avoid all
contact with the skin, mucosa, eyes and clothing.
6. All blood samples must be considered as potentially infectious and must be handled with care (in particular:
the wearing of protective gloves, gowns and goggles).
7. Blood tubes and disposable material used for handling should be disposed of in ad hoc containers intended
for incineration.
