Three-Dimensional Quantitative Phase Imaging for Characterizing Lymphocyte Subtypes
Transcript
Load a lymphocyte subtype suspension into an imaging chamber.
Position the chamber on the 3D quantitative phase imaging microscope stage.
Adjust microscope parameters and the focal plane for optimum imaging.
During lymphocyte imaging, the laser beam splits into reference and sample beams.
The Digital Micromirror Device, featuring a micromirror array in the sample beam's path, allows the beam to rotate 360 degrees around the optical axis.
Each beam passing through the lymphocyte undergoes a phase shift based on refractive index variations within the cell.
The resulting sample and reference beams recombine, forming a 2D hologram.
A sequence of holograms containing phase and amplitude information from different angles is captured through the beam rotation around the lymphocyte.
Acquire multiple background holograms with varying illumination angles, excluding lymphocytes.
Using the diffraction tomography algorithm, reconstruct the holograms into a 3D refractive index tomogram, providing morphological and biochemical information of lymphocytes without the need for labeling.
