Title:red blood cell suspension

Objective: to prepare 2%,5%,10% of cell suspension

Introduction :saline solution have used in immunohaematology lab for cell washing and to prepare cell suspension.1% sodium hpochlorite areprepared to clean up places contaimened by blood,suspensions of washed blood having and unknown antigens on their surface area are required for various test in the lab,the washing of cell is necessary to remove plasma ,which may interfere with the interferences with the reaction of the cells,in addition,blood group substances in plasma may neutralize the antiserum leading to false negative result,it also necessary to use red blood cell suspension of the appropriate strength for optimum reactionwith antibody molecule,the red blood cell are suspended in isotonic solution of the sodium chlorideat ph 7.0 adjusted

Procedure :

1. 1 drop of blood were put in the centrifuge tube

2. Added saline in it until there is 1cm left from the tube mouth

3. Then centrifuge it at 2500-3000 rpm for about 1-2 minutes

4. After centrifuge the supernatant are removed and blood are mixed well with another saline

5. The step 2-3 are repeated

6. When all the blood are done centrifuge prepare 2% suspension(1 drop blood&49 drop saline),5%(1drop blood,19 saline),10 % (1drop blood,9 drop saline)

Blood can be anticoagulated using a variety of reagents depending on what you wish to measure.

EDTA or CPA are common ones but no good if your assay requires complement.

If complement is needed then the blood must not be anticoagulated.

Regarding centrifugation of the primary sample, good separation is achieved with a spin speed of 2500rpm for 5 minutes.

Isotonic saline can be used as a diluent or PBS pH 7.0.

1 drop of whole blood in 49 of diluent, or 1 drop of packed red cells in 24 of diluent will give a 2% suspension.

I agree with thw answer give by Bashir Abdrhamn Bashir Mohamed.

Thanks!!!!for suggesting me.

Depending upon the purpose of experiment and research design , you have to collect blood in CPD or EDTA --> wash 3 times at 1500 rpm for 5 minutes with PBS and then, for example 100 ml 2% RBC suspension , take 2ml blood and add 98 ml PBS and make 2% RBC suspension of 100 ml. 

Best regards, 

Babu

Hi, after separation, prepare a 2 to 5 percent red blood

cell suspension for testing. There are several methods

that can be used for creating a cell suspension

preparation. Each laboratory should establish its own

guidelines for the preparation of cell suspensions.

Some suggested procedures include:

1. Using a disposable pipette, transfer approximately

0.2 ml (two drops) of the patients packed red blood

cells to a 10 x 75 mm test tube.

2. Fill the tube with saline. Cover and mix.

3. Centrifuge the tube at 3,400 rpm for 30 seconds,

then decant the saline.

4. Visually estimate a 2 to 5 percent suspension.

Adding approximately 1 ml of saline will make a 3

percent suspension, adding approximately 0.5 ml

of saline will make a 5 percent suspension.

Alternative Method For Preparation

Of 2 Percent Suspension

1. Place 1 to 2 ml of blood in large tube.

2. Fill with saline and centrifuge.

3. Aspirate/decant the remaining saline away from

the red blood cells.

4. Repeat washing (steps two and three) until supernatant

is clear. The remaining red blood cells are

called a packed red blood cell button.

5. Pipette 10 ml of saline into a clean test tube.

6. Add 0.2 ml (two drops) of the packed red blood cell

button to the 10 ml of saline. After the red blood

cells settle to the bottom of the test tube, rinse the

pipette with the surface layer of the saline to

remove any red blood cell residue.

ok great

any can give me answar how to make check cells

What temperature do you need to place the blood in the centrifuge?

I agree with Bashir Abdrhamn Bashir Mohamed and the preparation of 2% to 5% of blood by add 2 ml of packing blood to 100 ml of normal saline or PBS . this preparation was used within days only but their was other method can help to preserve RBCs for up to 4 year

Hello, what is the method to preserve RBCs for up to 4 year?