The diagnosis of leukemia involves the detection of the abnormal characteristics of blood cells by a trained pathologist. Currently, this is done manually by observing the morphological characteristics of white blood cells in the microscopic images. Though there are some equipment- based and chemical-based tests available, the use and adaptation of the automated computer vision-based system is still an issue. There are certain software frameworks available in the literature; however, they are still not being adopted commercially. So there is a need for an automated and software- based framework for the detection of leukemia. In software-based detection, segmentation is the first critical stage that outputs the region of interest for further accurate diagnosis. Therefore, this paper explores an efficient and hybrid segmentation that proposes a more efficient and effective system for leukemia diagnosis. A very popular publicly available database, the acute lymphoblastic leukemia image database (ALL-IDB), is used in this research. First, the images are pre-processed and segmentation is done using Multilevel thresholding with Otsu and Kapur methods. To further optimize the segmentation performance, the Learning enthusiasm-based teaching-learning-based optimization (LebTLBO) algorithm is employed. Different metrics are used for measuring the system performance. A comparative analysis of the proposed methodology is done with existing benchmarks methods. The proposed approach has proven to be better than earlier techniques with measuring parameters of PSNR and Similarity index. The result shows a significant improvement in the performance measures with optimizing threshold algorithms and the LebTLBO technique.