Anemia is a widespread health condition defined by reduced red blood cell (RBC) count or low hemoglobin (Hb) concentration, impairing oxygen delivery to tissues and organs. As per the World Health Organization (WHO), it is diagnosed when Hb levels fall below 12 g/dL in females and 13 g/dL in males. Common symptoms include fatigue, dizziness, weakness, headaches, and shortness of breath. Anemia is classified into microcytic, normocytic, and macrocytic types based on mean corpuscular volume (MCV). Diagnosis typically involves measuring Hb, hematocrit, and RBC indices such as MCV, mean cell hemoglobin concentration (MCHC), and reticulocyte count. Additional tests like peripheral blood smear (PBS) and hemoglobin electrophoresis help determine the underlying cause. However, conventional diagnostic methods often lack the ability to capture high-resolution, single-cell-level data, limiting their sensitivity to subtle RBC abnormalities. Assessing RBC structural and biochemical features provides deeper insights into anemia’s pathophysiology, enabling early detection, accurate classification, and improved clinical management. Against this background, the present prospective cross-sectional study aims to assess the relationship between iron levels and structural alteration of red blood cells using Quantitative phase Imaging in 24 individuals aged 20–30 years, including 12 anemic cases and 12 controls. Participants will be voluntarily recruited from individuals visiting Kasturba Hospital for routine health checkups, based on specified inclusion and exclusion criteria and after obtaining informed consent. Data will be statistically analyzed using Jamovi software, applying descriptive statistics (mean, median, standard deviation) and Pearson correlation. This study will enable identifying variations in structural alterations in red blood cells linked to anemia, while also investigating the fluctuation in iron level impact the structure and function of hemoglobin and enhancing the understanding of the biochemical connection between iron status and anemia.