ZOOL344 Lecture Notes - Lecture 6: Ocular Tonometry, Cuvette, Spectrophotometry
15 Nov 2017
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Experiment 1: spectral analysis of oxygenated and deoxygenated haemoglobin. Adjust spectrophotometer to 0 absorbance at lowest wavelength setting (590 nm) Add 6-8 ml of hb stolution (either ph 6. 8 or 7. 4) into cuvette. Put cuvette in separatory funnel portion of apparatus - avoid excess agitation! Increase wavelength by 10 nm increments up to 640 nm. Evacuate system by turning on vacuum pump (open stopcock #2) unti mercury manometer > 325 mmhg. Slowly open sc1 until mercruy stabilizes than close again. Manometer reading will change slightly as air is introduced from cuvette. Close sc2 to preserve vaccum in system and make sure sc1 is also closed. Introduce the solutions into the funnel portion and gently rotate while tipping back and forth for 8-10 mins = equilibrate solution to very low o2 concentration = note any colour change in pigment. Repeat proces to ensure hb is completely deoxygenated .
