Single-Frame Vignetting Correction for Post-Stitched-Tile Imaging Using VISTAmap

Stimulated Raman Scattering (SRS) nanoscopy imaging offers unprecedented insights into tissue molecular architecture but often requires stitching multiple high-resolution tiles to capture large fields of view. This process is time-consuming and frequently introduces vignetting artifacts-grid-like intensity fluctuations that degrade image quality and hinder downstream quantitative analyses and processing such as super-resolution deconvolution. We present VIgnetted Stitched-Tile Adjustment using Morphological Adaptive Processing (VISTAmap), a simple tool that corrects these shading artifacts directly on the final stitched image. VISTAmap automatically detects the tile grid configuration by analyzing intensity frequency variations and then applies sequential morphological operations to homogenize the image. In contrast to conventional approaches that require increased tile overlap or pre-acquisition background sampling, VISTAmap offers a pragmatic, post-processing solution without the need for separate individual tile images. This work addresses pressing concerns by delivering a robust, efficient strategy for enhancing mosaic image uniformity in modern nanoscopy, where the smallest details make tremendous impacts.