Background: Despite the wide range of methods available for measurement of haemoglobin (Hb), no single technique has emerged as the most appropriate and ideal for a blood donation setup. To cater to this need both invasive and non-invasive techniques of haemoglobin estimation were analysed. Aims and Objectives: To compare invasive and non-invasive methods of haemoglobin estimation in terms of accuracy, sensitivity and donor satisfaction. Materials and Methods: A prospective study utilizing 200 blood samples was carried out in a blood donation setting for quality evaluation of five methods of haemoglobin estimation: Haematology cell analyser (reference), DiaSpect, CuSO4, HemoCue and NBM-200. Results: Mean value of HemoCue (mean __ampersandsignplusmn; SD = 13.8 __ampersandsignplusmn; 1.7 g/dl) was higher by 0.26 compared to reference (mean __ampersandsignplusmn; SD = 13.54 __ampersandsignplusmn; 1.52 g/dl). DiaSpect proved to be the best technique (sensitivity 99.4%, specificity 94.4% and likelihood ratio 17.75). CuSO4 proved to be good with Negative Predictive Value close to 91.4%. NBM-200 shows wide variation but the mean of the difference being statistically not significant. Conclusion: CuSO4 method gives accurate results, if strict quality control is applied. HemoCue and DiaSpect are too expensive to be used as a primary screening method in an economically restricted country like India, but are accurate. NBM-200 is a non-invasive method and gives good result with better compliance and donor satisfaction.