SPLIT-PIN

SPLIT-PIN enhances optical sectioning in point-scanning microscopy by applying the SPLIT (separation of photons by lifetime tuning) algorithm to image series acquired with tunable pinhole sizes to optimize axial resolution while preserving signal-to-noise ratio.

Key Features:

• Optical Sectioning Enhancement: Applies the SPLIT (separation of photons by lifetime tuning) algorithm to image series acquired with tunable pinhole sizes to improve optical sectioning without closing the pinhole, preserving signal-to-noise ratio.
• Compatibility with Commercial Microscopes: Operates on data from microscopes equipped with tunable pinholes, including confocal and stimulated emission depletion (STED) systems.
• Image Processing Capabilities: Processes series of images acquired at different pinhole sizes to generate outputs with reduced out-of-focus background and enhanced image clarity.

Scientific Applications:

• Confocal Imaging: Improves confocal imaging and the visualization of DNA in non-adherent cell lines by reducing out-of-focus light.
• Super-Resolution STED Microscopy: Removes out-of-focus background in STED microscopy to improve image quality for sub-diffraction-limit structural analysis.
• Live Cell Imaging: Applies to live-cell imaging scenarios such as intestinal organoids stained with membrane dyes to maintain signal integrity during dynamic imaging.

Methodology:

Analyzes image series captured at various pinhole sizes and applies the SPLIT algorithm to separate photons based on their lifetimes, enabling enhanced optical sectioning and out-of-focus background reduction.