LAMBDA'S DAILY OPERATIONS
Find out more details about Lambda Neuroscience Foundation's laboratory operation, learn a bit more about histology, and explore what sets Lambda's method apart.
THE OVERALL PROCESS
Lambda will operate as a free, specialized histology service for neuroscience laboratories. Researchers conduct their experiments generating brain tissue samples within their laboratories, and then send those samples to Lambda similar to the process of medical doctors sending in patient biopsies for lab analysis. Lambda then performs the downstream sample processing needed to transform tissue into rigorous, highly granular digital image datasets that are indexed and available to researchers through Lambda’s dedicated database website. The goal is to provide researchers with a more information-rich view of the brain that is standardized across brain disease models to facilitate new molecular comparisons and insights, while reducing the technical and financial burden placed on individual laboratories.
WHAT HAPPENS IN THE LAB
Step 1: Prepare the slides by first using the Leica vibratome to make precise and reproducible brain sections starting from the front, slowly moving to the back. Then gently align the sections on the slide and allow them to air dry so they adhere to the surface of the slide glass.
Step 2: Incubate the brain sections on the slides first with a mixture of primary (1°) antibodies, each recognizing a specific structure in the tissue, followed by a mixture of secondary (2°) antibodies that label the structure/primary complex in a specific color (aka, a specific channel during imaging).
Step 3: Image the individual sections on the slides at high resolution on the Leica fluorescent microscope in multiple channels (7 total, 6 used for cellular identification).
DIGITAL IMAGE ANALYSIS & POST-PROCESSING
Channel signal provides information about tissue: Within a single channel, the cells making up the tissue can either be positive or negative. With two channels (red and green) it is possible to distinguish four cellular populations: R-/G- (double negative), R+/G- (red), R-/G+ (green), R+/G+ (double positive). Individual imaged tissue sections can be projected together using software to make a digital 3D rendering representing a volume of the tissue sample.
WHY LAMBDA'S METHOD IS SUPERIOR
Normal brain function strongly depends on properly constructed cellular architecture. Disease-related cellular and structural changes are highly informative of the molecular pathways affected by the disease, and therefore, what scientists can target to correct it. Histology (microscopic analysis of biological tissue) is one of the principal ways scientists determine how disease alters the brain because it allows visualization of molecular changes at the single-cell level. In many conventional workflows, however, histological analysis is limited to selected regions of the brain and relatively few imaging channels, which constrains how much information can be extracted from a given specimen. Lambda’s model is intentionally designed to expand both the scale and the detail of brain tissue visualization (see images labeled A and B for more detail).
Typically only 2 channels are used, allowing the researcher to identify 4 separate cellular populations. But with Lambda's method, up to 6 channels can be used allowing for up to 64 populations to be identified simultaneously (a 16x increase). By combining broader brain coverage with higher-dimensional imaging, the foundation aims to help researchers "see" brain diseases in an unprecedented and comprehensive way. This complete approach to whole brain visualization will lead to new insights and discoveries in brain disease progression, and will ultimately guide the development of potential new therapies.
