Abstract
AbstractIn this study, we compared the depth discrimination and speed performance of multifoci Raman hyperspectral imaging with the reference standard of a single laser point confocal Raman mapping. A liquid crystal spatial light modulator was employed for the generation of multifoci laser beams, and a digital micromirror device was used as a software‐configurable reflective pinhole array. The patterns of the laser foci and pinhole array can be rapidly changed without requiring any hardware alterations. Confocal patterns with different distance‐to‐size ratios were tested and compared. After optimization of the laser‐foci pattern, we demonstrated the feasibility of multifoci Raman hyperspectral microscopy for recording depth‐resolved molecular maps of biological cells (Acanthamoeba castellanii trophozoites). Micrometric depth discrimination and short acquisition times (20 min for single plane confocal image) were achieved.
Highlights
Raman microspectroscopy (RMS) is a powerful technique for highly specific molecular imaging of samples in three dimensions (3D).[1]
We have developed a technique based on spatial light modulators for multifoci Raman microscopy.[15]. This approach relies on a liquid crystal spatial light modulator (LC‐SLM) to generate a desired pattern of laser foci and employs a digital micromirror device (DMD) in the detection path to function as a software‐configurable reflective pinhole array
We have employed a multifoci Raman microscope based on a LC‐SLM for laser excitation and a DMD for confocal Raman detection to investigate the effect of the confocal period on depth discrimination
Summary
Raman microspectroscopy (RMS) is a powerful technique for highly specific molecular imaging of samples in three dimensions (3D).[1]. We have developed a technique based on spatial light modulators for multifoci Raman microscopy.[15] This approach relies on a liquid crystal spatial light modulator (LC‐SLM) to generate a desired pattern of laser foci and employs a digital micromirror device (DMD) in the detection path to function as a software‐configurable reflective pinhole array Because both the SLM and the DMD can be controlled through software, the positions of the laser foci can be rapidly changed without requiring any hardware alterations. The combination of LC‐SLM and DMD in the Raman microscope provided the flexibility to modify the number, location, and spacing between the laser foci in software without altering any optical components in the system This allows the changes in the recorded Raman hyperspectral images, such as depth discrimination with different 2D multifocal patterns to be investigated . After optimization of the laser foci pattern, we demonstrated the feasibility of recording depth‐resolved Raman maps of biological cells
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