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<title>4Pi microscope</title>

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<span id="openzim-page-title" class="mw-page-title-main"><span class="mw-page-title-main">4Pi microscope</span></span>

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<div id="mw-content-text" class="mw-body-content mw-content-ltr" lang="en" dir="ltr"><div class="mw-content-ltr mw-parser-output" lang="en" dir="ltr"><p>A <b>4Pi microscope</b> is a laser scanning <a href="Fluorescence_microscope" title="Fluorescence microscope">fluorescence microscope</a> with an improved <a href="Optical_axis" title="Optical axis">axial</a> <a href="Optical_resolution" title="Optical resolution">resolution</a>. With it the typical range of the axial resolution of 500–700 nm can be improved to 100–150 nm, which corresponds to an almost spherical focal spot with 5–7 times less volume than that of standard <a href="Confocal_microscopy" title="Confocal microscopy">confocal microscopy</a>.<sup id="cite_ref-1" class="reference"><a href="#cite_note-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup>

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<div class="mw-heading mw-heading2"><h2 id="Working_principle">Working principle</h2></div>

<p>The improvement in resolution is achieved by using two opposing objective lenses, which both are focused to the same geometrical location. Also the difference in <a href="Optical_path_length" title="Optical path length">optical path length</a> through each of the two <a href="Objective_lens" class="mw-redirect" title="Objective lens">objective lenses</a> is carefully aligned to be minimal. By this method, molecules residing in the common focal area of both objectives can be illuminated coherently from both sides and the reflected or emitted light can also be collected coherently, i.e. coherent superposition of emitted light on the detector is possible. The <a href="Solid_angle" title="Solid angle">solid angle</a> <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle \Omega }">

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<annotation encoding="application/x-tex">{\displaystyle \Omega }</annotation>

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</math></span><img src="./24b0d5ca6f381068d756f6337c08e0af9d1eeb6f.svg" class="mwe-math-fallback-image-inline mw-invert skin-invert" aria-hidden="true" style="vertical-align: -0.338ex; width:1.678ex; height:2.176ex;" alt="{\displaystyle \Omega }" loading="lazy"></span> that is used for illumination and detection is increased and approaches its maximum. In this case the sample is illuminated and detected from all sides simultaneously.

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<p>The operation mode of a 4Pi microscope is shown in the figure. The laser light is divided by a <a href="Beam_splitter" title="Beam splitter">beam splitter</a> and directed by mirrors towards the two opposing objective lenses. At the common focal point superposition of both focused light beams occurs. Excited molecules at this position emit fluorescence light, which is collected by both objective lenses, combined by the same beam splitter and deflected by a <a href="Dichroic_mirror" class="mw-redirect" title="Dichroic mirror">dichroic mirror</a> onto a detector. There, superposition of both emitted light pathways can take place again.

</p><p>In the ideal case each objective lens can collect light from a solid angle of <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle \Omega =2\pi }">

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<annotation encoding="application/x-tex">{\displaystyle \Omega =2\pi }</annotation>

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</math></span><img src="./ae7b014fa9ccbd6a4eb81eb9b42d9618b0550359.svg" class="mwe-math-fallback-image-inline mw-invert skin-invert" aria-hidden="true" style="vertical-align: -0.338ex; width:7.271ex; height:2.176ex;" alt="{\displaystyle \Omega =2\pi }" loading="lazy"></span>. With two objective lenses one can collect from every direction (solid angle <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle \Omega =4\pi }">

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<annotation encoding="application/x-tex">{\displaystyle \Omega =4\pi }</annotation>

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</math></span><img src="./40abe7ef17afd2715d78276e157fdd51c1c62800.svg" class="mwe-math-fallback-image-inline mw-invert skin-invert" aria-hidden="true" style="vertical-align: -0.338ex; width:7.271ex; height:2.176ex;" alt="{\displaystyle \Omega =4\pi }" loading="lazy"></span>). The name of this type of microscopy is derived from the maximal possible solid angle for excitation and detection. Practically, one can achieve only aperture angles of about 140° for an objective lens, which corresponds to <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle \Omega \approx 1.3\pi }">

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<annotation encoding="application/x-tex">{\displaystyle \Omega \approx 1.3\pi }</annotation>

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</math></span><img src="./3deb9f1706a3dcf7ba6adb740c955d21cc5e0e5b.svg" class="mwe-math-fallback-image-inline mw-invert skin-invert" aria-hidden="true" style="vertical-align: -0.338ex; width:9.08ex; height:2.176ex;" alt="{\displaystyle \Omega \approx 1.3\pi }" loading="lazy"></span>.

</p><p>The microscope can be operated in three different ways: In a 4Pi microscope of type A, the coherent superposition of excitation light is used to generate the increased resolution. The emission light is either detected from one side only or in an incoherent superposition from both sides. In a 4Pi microscope of type B, only the emission light is interfering. When operated in the type C mode, both excitation and emission light are allowed to interfere, leading to the highest possible resolution increase (~7-fold along the optical axis as compared to confocal microscopy).

</p><p>In a real 4Pi microscope light cannot be applied or collected from all directions equally, leading to so-called side lobes in the <a href="Point_spread_function" title="Point spread function">point spread function</a>. Typically (but not always) <a href="Two-photon_excitation_microscopy" title="Two-photon excitation microscopy">two-photon excitation microscopy</a> is used in a 4Pi microscope in combination with an emission pinhole to lower these side lobes to a tolerable level.

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<div class="mw-heading mw-heading2"><h2 id="History">History</h2></div>

<p>In 1971, <a href="Christoph_Cremer" title="Christoph Cremer">Christoph Cremer</a> and <a href="Thomas_Cremer" title="Thomas Cremer">Thomas Cremer</a> proposed the creation of a perfect <a href="Hologram" class="mw-redirect" title="Hologram">hologram</a>, i.e. one that carries the whole field information of the emission of a point source in all directions, a so-called <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle 4\pi }">

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<annotation encoding="application/x-tex">{\displaystyle 4\pi }</annotation>

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</math></span><img src="./057444bf35a0c22b19bcae1ef06e06ecdf8abe56.svg" class="mwe-math-fallback-image-inline mw-invert skin-invert" aria-hidden="true" style="vertical-align: -0.338ex; width:2.494ex; height:2.176ex;" alt="{\displaystyle 4\pi }" loading="lazy"></span> hologram.<sup id="cite_ref-2" class="reference"><a href="#cite_note-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-3" class="reference"><a href="#cite_note-3"><span class="cite-bracket">[</span>3<span class="cite-bracket">]</span></a></sup> However the publication from 1978 <sup id="cite_ref-4" class="reference"><a href="#cite_note-4"><span class="cite-bracket">[</span>4<span class="cite-bracket">]</span></a></sup> had drawn an improper physical conclusion (i.e. a point-like spot of light) and had completely missed the axial resolution increase as the actual benefit of adding the other side of the solid angle.<sup id="cite_ref-5" class="reference"><a href="#cite_note-5"><span class="cite-bracket">[</span>5<span class="cite-bracket">]</span></a></sup> The first description of a practicable system of 4Pi microscopy, i.e. the setup with two opposing, interfering lenses, was invented by <a href="Stefan_Hell" title="Stefan Hell">Stefan Hell</a> in 1991.<sup id="cite_ref-6" class="reference"><a href="#cite_note-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup> He demonstrated it experimentally in 1994.<sup id="cite_ref-7" class="reference"><a href="#cite_note-7"><span class="cite-bracket">[</span>7<span class="cite-bracket">]</span></a></sup>

</p><p>In the following years, the number of applications for this microscope has grown. For example, parallel excitation and detection with 64 spots in the sample simultaneously combined with the improved spatial resolution resulted in the successful recording of the dynamics of <a href="Mitochondria" class="mw-redirect" title="Mitochondria">mitochondria</a> in yeast cells with a 4Pi microscope in 2002.<sup id="cite_ref-8" class="reference"><a href="#cite_note-8"><span class="cite-bracket">[</span>8<span class="cite-bracket">]</span></a></sup> A commercial version was launched by microscope manufacturer <a href="Leica_Microsystems" title="Leica Microsystems">Leica Microsystems</a> in 2004<sup id="cite_ref-9" class="reference"><a href="#cite_note-9"><span class="cite-bracket">[</span>9<span class="cite-bracket">]</span></a></sup> and later discontinued.

</p><p>Up to now, the best quality in a 4Pi microscope was reached in conjunction with super-resolution techniques like the <a href="Stimulated_emission_depletion_microscope" class="mw-redirect" title="Stimulated emission depletion microscope">stimulated emission depletion</a> (STED) principle.<sup id="cite_ref-10" class="reference"><a href="#cite_note-10"><span class="cite-bracket">[</span>10<span class="cite-bracket">]</span></a></sup> Using a 4Pi microscope with appropriate excitation and de-excitation beams, it was possible to create a uniformly 50 nm sized spot, which corresponds to a decreased focal volume compared to confocal microscopy by a factor of 150–200 in fixed cells. With the combination of 4Pi microscopy and <a href="RESOLFT" title="RESOLFT">RESOLFT</a> microscopy with switchable proteins, it is now possible to take images of living cells at low light levels with isotropic resolutions below 40 nm.<sup id="cite_ref-11" class="reference"><a href="#cite_note-11"><span class="cite-bracket">[</span>11<span class="cite-bracket">]</span></a></sup>

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<ul><li><a href="Stimulated_emission_depletion_microscope" class="mw-redirect" title="Stimulated emission depletion microscope">Stimulated emission depletion microscope</a> (STED)</li>

<li><a href="Multifocal_plane_microscopy" title="Multifocal plane microscopy">Multifocal plane microscopy</a> (MUM)</li></ul>

<div class="mw-heading mw-heading2"><h2 id="References">References</h2></div>

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</style><cite id="CITEREFJ._BewersdorfA._EgnerS.W._Hell2004" class="citation journal cs1">J. Bewersdorf; A. Egner; S.W. Hell (2004). <a rel="nofollow" class="external text" href="http://www3.mpibpc.mpg.de/groups/hell/other_publications/GIT_4_04.pdf">"4Pi-Confocal Microscopy is Coming of Age"</a> <span class="cs1-format">(PDF)</span>. <i>GIT Imaging & Microscopy</i> (4): <span class="nowrap">24–</span>25.</cite></span>

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<li id="cite_note-2"><span class="mw-cite-backlink"><b><a href="#cite_ref-2">^</a></b></span> <span class="reference-text">Cremer C., Cremer T. (1971) <span class="mwe-math-element mwe-math-element-inline"><span class="mwe-math-mathml-inline mwe-math-mathml-a11y" style="display: none;"><math xmlns="http://www.w3.org/1998/Math/MathML" alttext="{\displaystyle 4\pi }">

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<annotation encoding="application/x-tex">{\displaystyle 4\pi }</annotation>

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<li id="cite_note-3"><span class="mw-cite-backlink"><b><a href="#cite_ref-3">^</a></b></span> <span class="reference-text">Considerations on a laser-scanning-microscope with high resolution and depth of field: C. Cremer and T. Cremer in MICROSCOPICA ACTA VOL. 81 NUMBER 1 September, p. 31–44 (1978). <a rel="nofollow" class="external text" href="http://www.kip.uni-heidelberg.de/AG_Cremer/pdf-files/Cremer_Micros_Acta_1978.pdf">Basic design of a confocal laser scanning fluorescence microscope & principle of a confocal laser scanning 4Pi fluorescence microscope, 1978</a> <a rel="nofollow" class="external text" href="https://web.archive.org/web/20160304030914/http://www.kip.uni-heidelberg.de/AG_Cremer/pdf-files/Cremer_Micros_Acta_1978.pdf">Archived</a> 2016-03-04 at the <a href="Wayback_Machine" title="Wayback Machine">Wayback Machine</a>.</span>

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<li id="cite_note-4"><span class="mw-cite-backlink"><b><a href="#cite_ref-4">^</a></b></span> <span class="reference-text">C. Cremer and T. Cremer (1978): Considerations on a laser-scanning-microscope with high resolution and depth of field Microscopica Acta VOL. 81 NUMBER 1 September, pp. 31—44 (1978) </span>

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<li id="cite_note-5"><span class="mw-cite-backlink"><b><a href="#cite_ref-5">^</a></b></span> <span class="reference-text">The Nobel Prize in Chemistry 2014 <a rel="nofollow" class="external free" href="https://www.nobelprize.org/prizes/chemistry/2014/hell/biographical/">https://www.nobelprize.org/prizes/chemistry/2014/hell/biographical/</a></span>

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<li id="cite_note-6"><span class="mw-cite-backlink"><b><a href="#cite_ref-6">^</a></b></span> <span class="reference-text">European patent <a rel="nofollow" class="external text" href="http://v3.espacenet.com/publicationDetails/biblio?CC=EP&NR=0491289">EP 0491289</a>.</span>

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<li id="cite_note-7"><span class="mw-cite-backlink"><b><a href="#cite_ref-7">^</a></b></span> <span class="reference-text"><cite id="CITEREFS._W._HellE._H._K._StelzerS._LindekC._Cremer1994" class="citation journal cs1">S. W. Hell; E. H. K. Stelzer; S. Lindek; C. Cremer (1994). <a rel="nofollow" class="external text" href="http://www.opticsinfobase.org/viewmedia.cfm?uri=ol-19-3-222&seq=0">"Confocal microscopy with an increased detection aperture: type-B 4Pi confocal microscopy"</a>. <i>Optics Letters</i>. <b>19</b> (3): <span class="nowrap">222–</span>224. <a href="Bibcode_(identifier)" class="mw-redirect" title="Bibcode (identifier)">Bibcode</a>:<a rel="nofollow" class="external text" href="https://ui.adsabs.harvard.edu/abs/1994OptL...19..222H">1994OptL...19..222H</a>. <a href="CiteSeerX_(identifier)" class="mw-redirect" title="CiteSeerX (identifier)">CiteSeerX</a> <span class="id-lock-free" title="Freely accessible"><a rel="nofollow" class="external text" href="https://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.501.598">10.1.1.501.598</a></span>. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1364%2FOL.19.000222">10.1364/OL.19.000222</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/19829598">19829598</a>.</cite></span>

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<li id="cite_note-8"><span class="mw-cite-backlink"><b><a href="#cite_ref-8">^</a></b></span> <span class="reference-text"><cite id="CITEREFA._EgnerS._JakobsS._W._Hell2002" class="citation journal cs1">A. Egner; S. Jakobs; S. W. Hell (2002). <a rel="nofollow" class="external text" href="http://www.pnas.org/content/99/6/3370.full.pdf">"Fast 100-nm resolution three-dimensional microscope reveals structural plasticity of mitochondria in live yeast"</a> <span class="cs1-format">(PDF)</span>. <i>PNAS</i>. <b>99</b> (6): <span class="nowrap">3370–</span>3375. <a href="Bibcode_(identifier)" class="mw-redirect" title="Bibcode (identifier)">Bibcode</a>:<a rel="nofollow" class="external text" href="https://ui.adsabs.harvard.edu/abs/2002PNAS...99.3370E">2002PNAS...99.3370E</a>. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<span class="id-lock-free" title="Freely accessible"><a rel="nofollow" class="external text" href="https://doi.org/10.1073%2Fpnas.052545099">10.1073/pnas.052545099</a></span>. <a href="PMC_(identifier)" class="mw-redirect" title="PMC (identifier)">PMC</a> <span class="id-lock-free" title="Freely accessible"><a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC122530">122530</a></span>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/11904401">11904401</a>.</cite></span>

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<li id="cite_note-9"><span class="mw-cite-backlink"><b><a href="#cite_ref-9">^</a></b></span> <span class="reference-text">Review article <a rel="nofollow" class="external text" href="http://www.kompetenznetze.de/netzwerke/opti-photonicnet/innovationshighlights/en/4Pi_Mikroskop">4Pi microscopy</a>.</span>

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<li id="cite_note-10"><span class="mw-cite-backlink"><b><a href="#cite_ref-10">^</a></b></span> <span class="reference-text"><cite id="CITEREFR._SchmidtC._A._WurmS._JakobsJ._Engelhardt2008" class="citation journal cs1">R. Schmidt; C. A. Wurm; S. Jakobs; J. Engelhardt; A. Egner; S. W. Hell (2008). "Spherical nanosized focal spot unravels the interior of cells". <i>Nature Methods</i>. <b>5</b> (6): <span class="nowrap">539–</span>544. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1038%2Fnmeth.1214">10.1038/nmeth.1214</a>. <a href="Hdl_(identifier)" class="mw-redirect" title="Hdl (identifier)">hdl</a>:<span class="id-lock-free" title="Freely accessible"><a rel="nofollow" class="external text" href="https://hdl.handle.net/11858%2F00-001M-0000-0012-DBBB-8">11858/00-001M-0000-0012-DBBB-8</a></span>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/18488034">18488034</a>. <a href="S2CID_(identifier)" class="mw-redirect" title="S2CID (identifier)">S2CID</a> <a rel="nofollow" class="external text" href="https://api.semanticscholar.org/CorpusID:16580036">16580036</a>.</cite></span>

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<li id="cite_note-11"><span class="mw-cite-backlink"><b><a href="#cite_ref-11">^</a></b></span> <span class="reference-text"><cite id="CITEREFU._BöhmS._W._HellR._Schmidt2016" class="citation journal cs1">U. Böhm; S. W. Hell; R. Schmidt (2016). <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4740410">"4Pi-RESOLFT nanoscopy"</a>. <i>Nature Communications</i>. <b>7</b> (10504): <span class="nowrap">1–</span>8. <a href="Bibcode_(identifier)" class="mw-redirect" title="Bibcode (identifier)">Bibcode</a>:<a rel="nofollow" class="external text" href="https://ui.adsabs.harvard.edu/abs/2016NatCo...710504B">2016NatCo...710504B</a>. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1038%2Fncomms10504">10.1038/ncomms10504</a>. <a href="PMC_(identifier)" class="mw-redirect" title="PMC (identifier)">PMC</a> <span class="id-lock-free" title="Freely accessible"><a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4740410">4740410</a></span>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/26833381">26833381</a>.</cite></span>

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</style><div id="Optical_microscopy454" style="font-size:114%;margin:0 4em"><a href="Microscopy" title="Microscopy">Optical microscopy</a></div></th></tr><tr><td class="navbox-abovebelow" colspan="3"><div>

<ul><li><b><a href="Microscope" title="Microscope">Microscope</a></b></li>

<li><b><a href="Optical_microscope" title="Optical microscope">Optical microscopy</a></b></li></ul>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Illumination and<br>contrast methods</th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Bright-field_microscopy" title="Bright-field microscopy">Bright-field microscopy</a></li>

<li><a href="K%C3%B6hler_illumination" title="Köhler illumination">Köhler illumination</a></li>

<li><a href="Dark-field_microscopy" title="Dark-field microscopy">Dark-field microscopy</a></li>

<li><a href="Phase-contrast_microscopy" title="Phase-contrast microscopy">Phase contrast</a></li>

<li><a href="Quantitative_phase-contrast_microscopy" title="Quantitative phase-contrast microscopy">Quantitative phase-contrast microscopy</a></li>

<li><a href="Differential_interference_contrast_microscopy" title="Differential interference contrast microscopy">Differential interference contrast (DIC)</a></li>

<li><a href="Dispersion_staining" title="Dispersion staining">Dispersion staining</a></li>

<li><a href="Second-harmonic_imaging_microscopy" title="Second-harmonic imaging microscopy">Second harmonic imaging (SHIM)</a></li>

<li><a href="Microscopy#Structured_illumination" title="Microscopy">Structured illumination</a></li>

<li><a href="Sarfus" title="Sarfus">Sarfus</a></li>

<li><a href="Interference_reflection_microscopy" title="Interference reflection microscopy">Interference reflection microscopy (IRM/RICM)</a></li>

<li><a href="Raman_microscope" title="Raman microscope">Raman</a></li></ul>

</div></td><td class="noviewer navbox-image" rowspan="3" style="width:1px;padding:0 0 0 2px"><div><span typeof="mw:File"></span></div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Fluorescence methods</th><td class="navbox-list-with-group navbox-list navbox-even" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Fluorescence_microscope" title="Fluorescence microscope">Fluorescence microscopy</a></li>

<li><a href="Confocal_microscopy" title="Confocal microscopy">Confocal microscopy</a></li>

<li><a href="Two-photon_excitation_microscopy" title="Two-photon excitation microscopy">Multiphoton microscopy</a> (<a href="Two-photon_excitation_microscopy" title="Two-photon excitation microscopy">Two-photon</a>, <a href="Three_photon_microscopy" class="mw-redirect" title="Three photon microscopy">Three-photon</a>)</li>

<li><a href="Deconvolution#Optics_and_other_imaging" title="Deconvolution">Image deconvolution</a></li>

<li><a href="Total_internal_reflection_fluorescence_microscope" title="Total internal reflection fluorescence microscope">Total internal reflection fluorescence microscopy (TIRF)</a></li>

<li><a href="Light_sheet_fluorescence_microscopy" title="Light sheet fluorescence microscopy">Lightsheet microscopy (LSFM/SPIM)</a></li>

<li><a href="Lattice_light-sheet_microscopy" title="Lattice light-sheet microscopy">Lattice light-sheet microscopy</a></li></ul>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Sub-diffraction<br>limit techniques</th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Diffraction-limited_system" title="Diffraction-limited system">Diffraction limit</a></li>

<li><a href="STED_microscopy" title="STED microscopy">Stimulated emission depletion (STED)</a></li>

<li><a href="Photoactivated_localization_microscopy" title="Photoactivated localization microscopy">Photo-activated localization microscopy (PALM/STORM)</a></li>

<li><a href="Near-field_scanning_optical_microscope" title="Near-field scanning optical microscope">Near-field (NSOM/SNOM)</a></li></ul>

</div></td></tr><tr><td class="navbox-abovebelow hlist" colspan="3"><div>

<ul><li><span class="noviewer" typeof="mw:File"><span title="Category"></span></span> <b>Category</b></li>

<li><span class="noviewer" typeof="mw:File"><span title="Commons page"></span></span> <b><a href="https://commons.wikimedia.org/wiki/Category:Optical_microscopy" class="extiw external" title="commons:Category:Optical microscopy">Commons</a></b></li></ul>

</div></td></tr></tbody></table></div></div><div><div class="zim-footer">

This article is issued from <a class="external text" title="Last edited on 2025-07-03" href="https://en.wikipedia.org/wiki/?title=4Pi_microscope&oldid=1298550288">Wikipedia</a>. The text is available under <a class="external text" href="https://creativecommons.org/licenses/by-sa/4.0/deed.en">Creative Commons Attribution-Share Alike 4.0</a> unless otherwise noted. Additional terms may apply for the media files.

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