In biological research, whole mount imaging allows for the comprehensive visualization of specimens, revolutionizing our grasp of complex biological structures and systems. Traditional two-dimensional imaging often misses the intricate details and spatial relationships crucial for understanding tissues, organs, and organisms. Three-dimensional (3D) reconstruction overcomes these limitations, offering enhanced visualization, analysis, and understanding of biological phenomena. This blog post highlights the benefits of 3D reconstruction, supported by a data-set acquired with Visikol’s confocal system at 20X magnification.
Clear Advantages of 3D Reconstruction
The paramount advantage of 3D reconstruction in whole mount imaging lies in the exceptional spatial awareness it provides. This is evident in our dataset, in which liver tissue was labeled with DAPI (nuclei), alpha-Smooth Muscle Actin (stellate cells), and Pan-collagen (extracellular matrix). Utilizing the reconstruction (Video 1), the complex interactions between the cellular and structural components of the liver are revealed in a way that traditional two-dimensional imaging methods cannot match. Planar techniques, while useful, often do not capture the intricate details and spatial relationships of these labeled components with the same clarity and depth. Structures that might be obscured or challenging to decipher in 2D slices (Video 2) are rendered clear and comprehensible in three dimension.
Video 1: Volumetric (3D) rendering of liver tissue, showcasing the DAPI, alpha-SMA, and Pan Collagen signals after background correction. Initially, the sample rotates from a top-down (xy) perspective to an orthogonal (yz) view. During each rotation, signals from individual markers are isolated, leading to a comprehensive composite that overlays all signals. A key in the top right corner identifies the stain signals being displayed.
Video 2: Planar (2D) view of liver tissue, showcasing the DAPI, alpha-SMA, and Pan Collagen signals after background correction. This perspective travels through the sample, one slice at a time in the z-direction (top-down), highlighting the overlap of all signals in each slice. A key in the bottom right corner identifies the stain signals being displayed.
Challenges You Might Not Have Considered
Venturing into the realm of 3D imaging, while enticing, brings its set of unexpected challenges. For example, dealing with thicker specimens introduces complications such as light scattering and absorption, which can significantly hinder light penetration depth and degrade image quality at deeper levels. To mitigate this, we employ confocal imaging techniques, which effectively reduce out-of-focus light but at the expense of longer acquisition times. Furthermore, the shift towards high-resolution imaging across large volumes results in the production of enormous data sets. Managing, storing, and processing this wealth of data demands substantial computational power and sophisticated data management strategies. The task of assembling thousands of individual images into a seamless 3D model is not only computationally intensive but also depends on advanced algorithms. Additionally, the progression towards 3D segmentation and signal quantification adds another layer of complexity, requiring even more computational resources and innovative solutions.
Why Visikol is Your Ally
Despite facing significant challenges, Visikol has achieved considerable advancements in crafting innovative tools, utilizing open-source resources to directly confront these issues. Our team specializes in developing bespoke algorithms designed to cater to the specific demands of 3D reconstruction and analysis. By refining computational models and incorporating sophisticated imaging techniques, Visikol is streamlining existing research approaches and unlocking the potential to investigate biological phenomena that were once beyond reach. This combination of technical prowess and inventive ingenuity places Visikol in a leading position to provide tailored solutions that meet the unique needs of our clients, further establishing our role as pioneers in the field.
If you’re interested in working with Visikol, please reach out to a member of our team today!
Author
Nicole Callen
Senior Image Analyst