Lab: Day 2

On the second day in the lab, we examined the amount of ferritin in the blood serum of our subjects and tested the newly developed ferritin test for the EUROlyser smart (Single Method Automated Reading Technology) 700 to see if the test could be done fast and effectively. The EUROlyser smart 700 (http://www.eurolyser.com) obtains a result in about five minutes and only required 50 µl of hemolytic serum. The test is supposed to be improved to where it only takes three minutes to get a result and requires only 20 µl of blood.

We borrowed two EUROlyser smart 700 and one of the inventors, Mr. Gruber, an engineer, came by personally to show us the steps involved and how to work the machine.

Small, but powerful.

Before we could put our samples into the machine, we first had to centrifuge the blood sample with 8000 spins every minute for five minutes.

Take my blood? Do you have to???

We then had to very carefully pipette out 50 µl of the serum. Next, the serum was placed into a test tube, which had been filled with MOPS-buffer and contained a small iron ball.

Reaction vessel for the EUROlyser.

After that, a reagent cap containing latex-stabilized antibodies is placed on top. The antibodies are highly specific in binding the ferritin, which causes cloudiness in the sample.

Through the changes in the cloudiness during the incubation time, one is able to draw conclusions about the amount of ferritin in the sample.

Now, one simply places the filled test tube into the EUROlyser smart 700 and presses the start button.

Blood can save lives

First, the machine warms the sample to exactly 37ºC. With the help of a magnet, the iron ball is moved in order to mix the sample with the buffer. Then the sample is incubated for 288 seconds.

The researcher at work

During this time, the antibodies attach themselves to the ferritin. The bonding of the antibodies is proportional to the concentration of ferritin in the sample. The bond is measured automatically by a photometer. The instrument records the entire curve and from it, calculates the amount of ferritin.

The higher the bond, then the less light is able to shine through. The amount of light that penetrates through is then measured in mOD/min and gives a measurement.

Does the pricking really not hurt???

Result:

With ferritin, one finds as limit values

150µg/l (ng/ml) Women before menopause

250µg/l (ng/ml) Men, Women after menopause

Sample ID Result
1 135 ng/ml
2 77 ng/ml
3 122 ng/ml
4 162 ng/ml
5 77 ng/ml
6 10 ng/ml
7 48 ng/ml
8 274 ng/ml
9 185 ng/ml
10 418 ng/ml
11 33 ng/ml
12 80 ng/ml
13 2 ng/ml
14 139 ng/ml
15 88 ng/ml
16 111 ng/ml
17 26 ng/ml
18 138 ng/ml
19 62 ng/ml
20 230 ng/ml
21 0 ng/ml
22 2 ng/ml
23 35 ng/ml
24 143 ng/ml
25 38 ng/ml
26 45 ng/ml
27 148 ng/ml
28 110 ng/ml
29 81 ng/ml
30 247 ng/ml
31 297 ng/ml
32 88 ng/ml
33 8 ng/ml
34 35 ng/ml
35 64 ng/ml
36 0 ng/ml
37 220 ng/ml
38 11 ng/ml
39 22 ng/ml
40 0 ng/ml

A result displayed graphically

41 395 ng/ml
42 62 ng/ml
43 120 ng/ml
44 14 ng/ml
45 0 ng/ml
46 66 ng/ml
47 66 ng/ml
48 133 ng/ml
49 74 ng/ml
50 110 ng/ml
51 175 ng/ml
52 0 ng/ml
53 69 ng/ml
54 94 ng/ml