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<p>MINFLUX fluorescence microscopy is a recently introduced super-resolution approach for studying cellular structures and their dynamics with highest detail. MINFLUX (<i>MFX</i>) performs Single Particle Tracking (SPT) at <i>runtime</i>. The ad hoc signal interpretation necessary to sustain the method relies on several parameters, which need to be optimized in relation to the sample under study, such as fluorescent lipid analogs in membranes, to ensure the fidelity of the measurement. We propose a parameter optimization strategy, give&#xa0;an overview of the most important parameters, present a theoretical upper limit for trackable diffusion rates, and demonstrate <i>MFX</i>-enabled SPT of fast <InlineEquation ID="IEq1"> <InlineMediaObject> <ImageObject Color="BlackWhite" FileRef="42003_2025_9060_Article_IEq1.gif" Format="GIF" Height="23" Rendition="HTML" Resolution="72" Type="Linedraw" Width="162" /> </InlineMediaObject> <EquationSource Format="TEX">\(\left(\left\langle {D}_{{{\rm{MSD}}}}\right\rangle =2.5\mu {m}^{2}/s\right)\)</EquationSource> <EquationSource Format="MATHML"><math> <mfenced close=")" open="("> <mrow> <mfenced close="⟩" open="⟨"> <mrow> <msub> <mrow> <mi>D</mi> </mrow> <mrow> <mi mathvariant="normal">MSD</mi> </mrow> </msub> </mrow> </mfenced> <mo>=</mo> <mn>2.5</mn> <mi>μ</mi> <msup> <mrow> <mi>m</mi> </mrow> <mrow> <mn>2</mn> </mrow> </msup> <mo>/</mo> <mi>s</mi> </mrow> </mfenced> </math></EquationSource> </InlineEquation> lateral Brownian motion of lipids in membranes.</p>

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Parameter optimization for MINFLUX microscopy enabled single particle tracking

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月14日

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  • Bela T. L. Vogler,
  • Giovanni De Angelis,
  • Ziliang Zhao,
  • Christian Eggeling,
  • Francesco Reina

摘要

MINFLUX fluorescence microscopy is a recently introduced super-resolution approach for studying cellular structures and their dynamics with highest detail. MINFLUX (MFX) performs Single Particle Tracking (SPT) at runtime. The ad hoc signal interpretation necessary to sustain the method relies on several parameters, which need to be optimized in relation to the sample under study, such as fluorescent lipid analogs in membranes, to ensure the fidelity of the measurement. We propose a parameter optimization strategy, give an overview of the most important parameters, present a theoretical upper limit for trackable diffusion rates, and demonstrate MFX-enabled SPT of fast \(\left(\left\langle {D}_{{{\rm{MSD}}}}\right\rangle =2.5\mu {m}^{2}/s\right)\) D MSD = 2.5 μ m 2 / s lateral Brownian motion of lipids in membranes.