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Inline calibration of spatial light modulators in nonlinear microscopy.

Daniël W S Cox, Harish Sasikumar, Ivo M Vellekoop

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    Summary
    This summary is machine-generated.

    We developed a new calibration method for phase-only spatial light modulators in nonlinear microscopy. This technique uses microscope images, eliminating the need for extra hardware and working well in low-light conditions.

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    Area of Science:

    • Optics and Photonics
    • Microscopy
    • Biophysics

    Background:

    • Phase-only spatial light modulators (SLMs) are crucial for advanced microscopy techniques.
    • Accurate calibration of SLMs is essential for precise control of light wavefronts.
    • Existing calibration methods often require specialized hardware or complex procedures.

    Purpose of the Study:

    • To present a novel, hardware-independent method for calibrating phase-only SLMs in nonlinear microscopy.
    • To adapt SLM calibration to the specific nonlinear signals generated in multi-photon excitation fluorescence microscopy.
    • To demonstrate the robustness of the method under challenging imaging conditions.

    Main Methods:

    • The proposed method utilizes the microscope's own image as the calibration measurement.
    • It directly analyzes the nonlinear signals generated during multi-photon excitation.
    • No additional optical components or external hardware are required for calibration.

    Main Results:

    • The method successfully calibrates the phase-only SLM response within the nonlinear microscopy setup.
    • Effective calibration was achieved even under low-light imaging conditions.
    • The technique demonstrated resilience against strong photobleaching effects.

    Conclusions:

    • This image-based calibration method offers a simple and efficient solution for phase-only SLMs in nonlinear microscopy.
    • It significantly reduces the complexity and cost associated with SLM calibration.
    • The approach is suitable for routine use in multi-photon fluorescence microscopy, even with demanding samples.