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2. OBLICZENIA |
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Przygotowanie masy erytrocytarnej o hematokrycie 3% |
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początkowy hematokryt: |
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H= 71% |
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C1V1 = C2V2 |
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71% V1 = 3% * 17 ml |
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VNaCl = 17 ml - 0,7183 ml = 16,2817 ml |
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Pobranie 0,7183 ml krwi. Przeniesienie do 8 probówek po 2ml masy erytrocytarnej i dodanie ftalocyjaniny |
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do C= 3*10-6M dysponując C= 3*10-4 M |
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C1V1 = C2V2 |
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3*10-4 V2= 3*10-6 * 2ml |
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V2 = 0,02 ml = 20µl |
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- objętość ftalocyjaniny |
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Inkubowanie sporządzonych próbek (próby badane i kontrolne-bez ftalocyjaniny) w ciemności(20min) |
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Naświetlanie próbek promieniowaniem laserowym o długości fali = 670nm kolejno przez 2, 5 i 10 min przy |
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jednoczesnym mieszaniu na mieszadle magnetycznym. Po naświetlaniu odwirowywano krwinki przez |
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10 min przy 3000 obrotów/ min. |
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Oznaczenie zawartości hemoglobiny w otrzymanym supernatancie przez pomiar absorbancji przy λ=540 nm. |
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CZAS |
PRÓBKI Z Pht |
PRÓBKI BEZ Pht |
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kontrola |
0.182 |
0.146 |
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2 min |
0.222 |
0.058 |
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5min |
0.401 |
0.115 |
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10 min |
0.603 |
0.113 |
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bez Pth |
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z Pth |
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większą wartość absorbancji niż bez Pht. Dodanie Pht do masy erytrocytarnej i działanie laserem |
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zwiększyło znacznie jej absorbancję. |
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