Introduction: Plant micromorphology provides essential morphological evidence for species identification, functional trait analysis, and evolutionary studies. However, acquiring high-quality micromorphological images has traditionally required expensive optical microscopes and complex sample preparation, which limits accessibility for small- to medium-sized laboratories and educational settings. Recently, macro focus stacking—a photographic technique widely used to capture high-depth-of-field images—has emerged as a low-cost alternative for high-resolution imaging. Despite its potential, systematic workflows optimized for plant specimens remain scarce. Rationale: To address these limitations, we developed a cost-effective and efficient imaging workflow tailored for plant micromorphology. The system integrates commercially available components, including long working distance microscope objectives, a full-frame digital camera, a motorized focusing rail, and professional image-stacking software. The workflow is designed to: overcome the shallow depth-of-field inherent in high-magnification imaging by sequentially capturing and fusing focal planes; Preserve the natural color and morphology of plant tissues; Avoid complex sample preparation, thereby minimizing preparation-induced artifacts such as surface shrinkage or structural collapse. The workflow encompasses sample collection and minimal preparation, system assembly, sequential focal-plane acquisition, image stacking and fusion, and scale calibration. The total hardware cost ranges approximately from 10 000 to 50 000 CNY, making the system accessible to small- and medium-sized laboratories without compromising imaging quality. Results: Evaluation using USAF 1951 resolution targets demonstrated that the system achieves a practical resolution of 0.87–2.46 μm, depending on the optical configuration, imaging conditions, and the numerical aperture of the objective lens. This resolution allows stable visualization of micron-scale structures, including epidermal cell contours, trichome morphology, primary ornamentation of pollen exines, and seed surface sculptures. Compared with conventional optical microscopy, focus-stacking substantially extends the depth of field, enabling full-focus imaging of structurally complex, three-dimensional surfaces that cannot be captured in a single frame. The system operates under ambient conditions, preserves true-to-life color, and avoids common preparation artifacts. Its low hardware cost, operational flexibility, and ease of assembly make it particularly suitable for small- and medium-sized plant biology laboratories. Conclusion: We present a practical, low-cost macro focus stacking workflow for plant micromorphological imaging that delivers micron-scale resolution, extended depth of field, and artifact-free, color-accurate images. The method is well-suited for taxonomic and morphological studies, botanical education, virtual herbarium construction, and science outreach. By combining affordable hardware with a streamlined workflow, this system provides an efficient and accessible solution for high-quality imaging of plant organs and cellular structures, bridging the gap between conventional light microscopy and high-resolution morphological analysis without the need for complex sample preparation.
Chang et al. (Fri,) studied this question.