fixative agent. It is provided as a 10X concentrate.

A
working solution (1X) of this concentrate is to be
used for the fixation of cell preparations before
analysis by flow cytometry.

ReaFix Fixative Solution Note - Follow appropriate instructions as

10X Concentrate given in the respective product inserts for
information on instruments, sample collection and
Catalog No. 25325- 00 sample processing.

1. PRODUCT DESCRIPTION 4.1 Dilution Instructions: Preparation of the

1X Working Solution
The ReaFix fixative solution is a 10X
concentrated formaldehyde based fixative solution in Dilute the 10X concentrate 1:10 deionized
Phosphate buffer saline (PBS). It is intended for water at room temperature (20° to 25°C) to prepare
fixing cell preparations to enable a fix-no wash 1X working fixative solution according to Table 1.
protocol in applications that involve Mix well.
immunofluorescent staining and analysis by flow Prepare an adequate amount of the 1X
cytometry. working solution depending on the reagent and
volume to be used per tube.
2. INTENDED USE
Table 1: Dilution Instructions for Preparation of 1X
The ReaFix fixative solution is recommended Working Fixative solution
for use in fixation of cell preparations stained with
ReaMetrix reagents such as Rea T-Count reagent Required Volume
(Dual Tube Catalog Number: 25124-00; Single Tube
of 1X Working 5 10 15 20 25
Catalog Number: 25242-00) and ReaPan Thrombo
solution (mL)
reagent (Catalog Number: 25232-00) which are to be
Volume of 10X
analyzed by flow cytometry.
Use of ReaFix fixative solution in the Concentrate
0.5 1.0 1.5 2.0 2.5
processing steps for analysis of the sample ReaFix Fixative
preparations to be analyzed using the above- Solution (mL)
mentioned reagents (Section 1) is highly Volume of
recommended. This ensures the sufficient fixing of Deionized Water 4.5 9.0 13.5 18.0 22.5
the immunofluorescent stained populations without (mL)
lysing the red blood cells

3. PRINCIPLE 4.2 Storage Condition and Stability

When whole blood is added to the
monoclonal antibody reagent, the fluorochrome-
labeled antibodies in the reagent bind specifically to
target surface antigens. The stained samples are then
treated with ReaFix fixative solution which stabilizes
cell proteins by binding covalently between the free
amine groups, without lysing the red blood cells. The
stained fixed cell preparations are suitable for flow
cytometric analysis.
4. REAGENT
The 1X working fixative solution should be prepared
daily; the unused 1X Fixative solution should be
discarded at the end of the day. Do not refrigerate.
4.3 Evidence of Deterioration
The normal appearance of ReaFix Fixative Solution
reagent is that of a clear liquid. Any change in the
physical appearance of the reagent, major variation in
values for control subjects may indicate deterioration,
and the reagent should not be used.

The ReaFix fixative solution is formulated as

a buffered solution and contains Formaldehyde as a
5. PRECAUTIONS
1. The ReaFix Fixative Solution contains
Formaldehyde. Formaldehyde is a potential
carcinogen and causes irritation to the eyes and
skin. Avoid inhalation, contact with eyes, skin and
clothing. Wear suitable protective clothing. Never
pipette by mouth; if swallowed, seek medical
attention immediately.
2. All blood specimens are considered biohazards.
Handle them as if they are capable of transmitting
infection and dispose off with proper precautions
in accordance with governmental regulations.
3. The addition of precise volume of deionized water
is critical to obtain correct concentration of the 1X
working solution. Use a calibrated pipette and
operate according to the manufacturer’s
instructions.
4. The use of temperatures, incubation and vortexing
times other than those specified in the
corresponding product inserts may give erroneous
results.
5. The ReaFix Fixative Solution contains
Formaldehyde which is highly volatile. Do not
leave the lid in a loose or open position. The lid
should be tightly sealed after use.
6. Avoid microbial contamination of the reagent or
erroneous results may occur.
7. Results obtained with flow cytometry may be
erroneous if the instrument is misaligned,
compensation spillovers are not correctly
compensated or the gate is improperly set.
8. Reproducible results will be obtained as long as
the procedure used is in accordance with this
package insert and with good laboratory practice
guidelines.
Immunophenotypic Techniques. Current
Protocols in Cytometry (2000) 6.8.1-6.8.26.
3. Bossuyt X, Marti GE, Fleisher TA:
Comparative analysis of whole blood lysis
methods for flow cytometry. Cytometry
30:124–133, 1997.
4. Dressler, L.G., "Specimen handling, storage,
and preparation", 1997, Curr. Protocols
Cytometry, Chapter 5, 5.0.1- 5.2.15.
7. WARRANTY
The product is warranted only to conform to
the quantity and contents stated on the label at the
time of delivery to the customer. There are no
warranties, expressed or implied, that extend beyond
the description on the label of the product.
ReaMetrix’ sole liability is limited to replacement of
the products. ReaMetrix is not liable for property
damage, personal injury, or economic loss caused by
the product.
CUSTOMER SUPPORT INFORMATION
ReaMetrix India Pvt. Ltd.
50-B, II Phase, Peenya Industrial Area
Peenya, Bangalore-560058, India
Ph: 91-80-28378693/5,
Fax: 91-80-41172451
E-mail: info@reametrix.com
www.reametrix.com
Rev No. 1.0, 27-Apr-09

6. REFERENCES

1. Mandy FF, Nicholson JK, McDougal JS.

Guidelines for performing single-platform
absolute CD4+ T-cell determinations with
CD45 gating for persons infected with
human immunodeficiency virus. Centers for
Disease Control and Prevention. MMWR
Recomm Rep 2003 Jan 31; 52(RR-2):1-13.

2. Mandy FF, Brando B. Enumeration of

Absolute Cell Counts Using