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Available in 22 languages" > <label id="p-lang-btn-label" for="p-lang-btn-checkbox" class="vector-dropdown-label cdx-button cdx-button--fake-button cdx-button--fake-button--enabled cdx-button--weight-quiet cdx-button--action-progressive mw-portlet-lang-heading-22" aria-hidden="true" ><span class="vector-icon mw-ui-icon-language-progressive mw-ui-icon-wikimedia-language-progressive"></span> <span class="vector-dropdown-label-text">22 languages</span> </label> <div class="vector-dropdown-content"> <div class="vector-menu-content"> <ul class="vector-menu-content-list"> <li class="interlanguage-link interwiki-ar mw-list-item"><a href="https://ar.wikipedia.org/wiki/%D8%AA%D8%A3%D9%84%D9%82_%D9%85%D9%86%D8%A7%D8%B9%D9%8A" title="تألق مناعي – Arabic" lang="ar" hreflang="ar" data-title="تألق مناعي" data-language-autonym="العربية" data-language-local-name="Arabic" class="interlanguage-link-target"><span>العربية</span></a></li><li class="interlanguage-link interwiki-bs mw-list-item"><a href="https://bs.wikipedia.org/wiki/Imunoflurescencija" title="Imunoflurescencija – Bosnian" lang="bs" hreflang="bs" data-title="Imunoflurescencija" data-language-autonym="Bosanski" data-language-local-name="Bosnian" class="interlanguage-link-target"><span>Bosanski</span></a></li><li class="interlanguage-link interwiki-ca mw-list-item"><a href="https://ca.wikipedia.org/wiki/Immunofluoresc%C3%A8ncia" title="Immunofluorescència – Catalan" lang="ca" hreflang="ca" data-title="Immunofluorescència" data-language-autonym="Català" data-language-local-name="Catalan" class="interlanguage-link-target"><span>Català</span></a></li><li class="interlanguage-link interwiki-cs mw-list-item"><a href="https://cs.wikipedia.org/wiki/Imunofluorescence" title="Imunofluorescence – Czech" lang="cs" hreflang="cs" data-title="Imunofluorescence" data-language-autonym="Čeština" data-language-local-name="Czech" class="interlanguage-link-target"><span>Čeština</span></a></li><li class="interlanguage-link interwiki-de mw-list-item"><a href="https://de.wikipedia.org/wiki/Immunfluoreszenz" title="Immunfluoreszenz – German" lang="de" hreflang="de" data-title="Immunfluoreszenz" data-language-autonym="Deutsch" data-language-local-name="German" class="interlanguage-link-target"><span>Deutsch</span></a></li><li class="interlanguage-link interwiki-et mw-list-item"><a href="https://et.wikipedia.org/wiki/Immunofluorestsents" title="Immunofluorestsents – Estonian" lang="et" hreflang="et" data-title="Immunofluorestsents" data-language-autonym="Eesti" data-language-local-name="Estonian" class="interlanguage-link-target"><span>Eesti</span></a></li><li class="interlanguage-link interwiki-es mw-list-item"><a href="https://es.wikipedia.org/wiki/Inmunofluorescencia" title="Inmunofluorescencia – Spanish" lang="es" hreflang="es" data-title="Inmunofluorescencia" data-language-autonym="Español" data-language-local-name="Spanish" class="interlanguage-link-target"><span>Español</span></a></li><li class="interlanguage-link interwiki-eu mw-list-item"><a href="https://eu.wikipedia.org/wiki/Immunofluoreszentzia" title="Immunofluoreszentzia – Basque" lang="eu" hreflang="eu" data-title="Immunofluoreszentzia" data-language-autonym="Euskara" data-language-local-name="Basque" class="interlanguage-link-target"><span>Euskara</span></a></li><li class="interlanguage-link interwiki-fr mw-list-item"><a href="https://fr.wikipedia.org/wiki/Immunofluorescence" title="Immunofluorescence – French" lang="fr" hreflang="fr" data-title="Immunofluorescence" data-language-autonym="Français" data-language-local-name="French" class="interlanguage-link-target"><span>Français</span></a></li><li class="interlanguage-link interwiki-gl mw-list-item"><a href="https://gl.wikipedia.org/wiki/Inmunofluorescencia" title="Inmunofluorescencia – Galician" lang="gl" hreflang="gl" data-title="Inmunofluorescencia" data-language-autonym="Galego" data-language-local-name="Galician" class="interlanguage-link-target"><span>Galego</span></a></li><li class="interlanguage-link interwiki-ko mw-list-item"><a href="https://ko.wikipedia.org/wiki/%EB%A9%B4%EC%97%AD_%ED%98%95%EA%B4%91%EB%B2%95" title="면역 형광법 – Korean" lang="ko" hreflang="ko" data-title="면역 형광법" data-language-autonym="한국어" data-language-local-name="Korean" class="interlanguage-link-target"><span>한국어</span></a></li><li class="interlanguage-link interwiki-hi mw-list-item"><a href="https://hi.wikipedia.org/wiki/%E0%A4%87%E0%A4%AE%E0%A5%8D%E0%A4%AF%E0%A5%82%E0%A4%A8%E0%A5%8B%E0%A4%AB%E0%A5%8D%E0%A4%B2%E0%A5%8B%E0%A4%B0%E0%A5%87%E0%A4%B8%E0%A5%87%E0%A4%82%E0%A4%B8" title="इम्यूनोफ्लोरेसेंस – Hindi" lang="hi" hreflang="hi" data-title="इम्यूनोफ्लोरेसेंस" data-language-autonym="हिन्दी" data-language-local-name="Hindi" class="interlanguage-link-target"><span>हिन्दी</span></a></li><li class="interlanguage-link interwiki-id mw-list-item"><a href="https://id.wikipedia.org/wiki/Imunofluoresen" title="Imunofluoresen – Indonesian" lang="id" hreflang="id" data-title="Imunofluoresen" data-language-autonym="Bahasa Indonesia" data-language-local-name="Indonesian" class="interlanguage-link-target"><span>Bahasa Indonesia</span></a></li><li class="interlanguage-link interwiki-it mw-list-item"><a href="https://it.wikipedia.org/wiki/Immunofluorescenza" title="Immunofluorescenza – Italian" lang="it" hreflang="it" data-title="Immunofluorescenza" data-language-autonym="Italiano" data-language-local-name="Italian" class="interlanguage-link-target"><span>Italiano</span></a></li><li class="interlanguage-link interwiki-hu mw-list-item"><a href="https://hu.wikipedia.org/wiki/Immunfluoreszcencia" title="Immunfluoreszcencia – Hungarian" lang="hu" hreflang="hu" data-title="Immunfluoreszcencia" data-language-autonym="Magyar" data-language-local-name="Hungarian" class="interlanguage-link-target"><span>Magyar</span></a></li><li class="interlanguage-link interwiki-ja mw-list-item"><a href="https://ja.wikipedia.org/wiki/%E8%9B%8D%E5%85%89%E6%8A%97%E4%BD%93%E6%B3%95" title="蛍光抗体法 – Japanese" lang="ja" hreflang="ja" data-title="蛍光抗体法" data-language-autonym="日本語" data-language-local-name="Japanese" class="interlanguage-link-target"><span>日本語</span></a></li><li class="interlanguage-link interwiki-pl mw-list-item"><a href="https://pl.wikipedia.org/wiki/Test_immunofluorescencyjny" title="Test immunofluorescencyjny – Polish" lang="pl" hreflang="pl" data-title="Test immunofluorescencyjny" data-language-autonym="Polski" data-language-local-name="Polish" class="interlanguage-link-target"><span>Polski</span></a></li><li class="interlanguage-link interwiki-pt mw-list-item"><a href="https://pt.wikipedia.org/wiki/Imunofluoresc%C3%AAncia" title="Imunofluorescência – Portuguese" lang="pt" hreflang="pt" data-title="Imunofluorescência" data-language-autonym="Português" data-language-local-name="Portuguese" class="interlanguage-link-target"><span>Português</span></a></li><li class="interlanguage-link interwiki-ro mw-list-item"><a href="https://ro.wikipedia.org/wiki/Imunofluorescen%C8%9B%C4%83" title="Imunofluorescență – Romanian" lang="ro" hreflang="ro" data-title="Imunofluorescență" data-language-autonym="Română" data-language-local-name="Romanian" class="interlanguage-link-target"><span>Română</span></a></li><li class="interlanguage-link interwiki-ru mw-list-item"><a href="https://ru.wikipedia.org/wiki/%D0%98%D0%BC%D0%BC%D1%83%D0%BD%D0%BE%D1%84%D0%BB%D1%83%D0%BE%D1%80%D0%B5%D1%81%D1%86%D0%B5%D0%BD%D1%82%D0%BD%D1%8B%D0%B9_%D0%B0%D0%BD%D0%B0%D0%BB%D0%B8%D0%B7" title="Иммунофлуоресцентный анализ – Russian" lang="ru" hreflang="ru" data-title="Иммунофлуоресцентный анализ" data-language-autonym="Русский" data-language-local-name="Russian" class="interlanguage-link-target"><span>Русский</span></a></li><li class="interlanguage-link interwiki-sv mw-list-item"><a href="https://sv.wikipedia.org/wiki/Immunofluorescens" title="Immunofluorescens – Swedish" lang="sv" hreflang="sv" data-title="Immunofluorescens" 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dir="ltr"><div class="shortdescription nomobile noexcerpt noprint searchaux" style="display:none">Technique used for light microscopy</div> <figure typeof="mw:File/Thumb"><a href="/wiki/File:Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/a/af/Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg/278px-Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg" decoding="async" width="278" height="251" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/a/af/Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg/417px-Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/a/af/Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg/556px-Blood_vessels_in_porcine_skin_-_SMA_A488_-_20x.jpg 2x" data-file-width="2042" data-file-height="1843" /></a><figcaption>Vasculature of porcine <a href="/wiki/Skin" title="Skin">skin</a> under fluorescence (<a href="/wiki/Smooth_muscle" title="Smooth muscle">Smooth muscle</a> <a href="/wiki/Actin" title="Actin">actin</a> with AlexaFluor 488). Green = smooth muscle actin (SMA) with Alexa 488 fluorophore. Blue = DAPI counterstain. Red = auto-fluorescence. </figcaption></figure> <p> <b>Immunofluorescence</b> <b>(IF)</b> is a <a href="/wiki/Optical_microscope" title="Optical microscope">light microscopy</a>-based technique that allows detection and localization of a wide variety of target <a href="/wiki/Biomolecule" title="Biomolecule">biomolecules</a> within a cell or tissue at a quantitative level. The technique utilizes the binding specificity of <a href="/wiki/Antibodies" class="mw-redirect" title="Antibodies">antibodies</a> and <a href="/wiki/Antigen" title="Antigen">antigens</a>.<sup id="cite_ref-:0_1-0" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> The specific region an antibody recognizes on an antigen is called an <a href="/wiki/Epitope" title="Epitope">epitope</a>. Several antibodies can recognize the same epitope but differ in their binding affinity. The antibody with the higher affinity for a specific epitope will surpass antibodies with a lower affinity for the same epitope.<sup id="cite_ref-:2_2-0" class="reference"><a href="#cite_note-:2-2"><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> </p><p>By conjugating the antibody to a <a href="/wiki/Fluorophore" title="Fluorophore">fluorophore</a>, the position of the target biomolecule is visualized by exciting the fluorophore and measuring the emission of light in a specific predefined wavelength using a <a href="/wiki/Fluorescence_microscope" title="Fluorescence microscope">fluorescence microscope</a>. It is imperative that the binding of the <a href="/wiki/Fluorophore" title="Fluorophore">fluorophore</a> to the antibody itself does not interfere with the immunological specificity of the antibody or the binding capacity of its antigen.<sup id="cite_ref-:3_4-0" class="reference"><a href="#cite_note-:3-4"><span class="cite-bracket">[</span>4<span class="cite-bracket">]</span></a></sup><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> </p><p> Immunofluorescence is a widely used example of <a href="/wiki/Immunostaining" title="Immunostaining">immunostaining</a> (using antibodies to stain proteins) and is a specific example of <a href="/wiki/Immunohistochemistry" title="Immunohistochemistry">immunohistochemistry</a> (the use of the antibody-antigen relationship in tissues). This technique primarily utilizes fluorophores to visualize the location of the antibodies, while others provoke a color change in the environment containing the antigen of interest or make use of a radioactive label. Immunofluorescent techniques that utilized labelled antibodies was conceptualized in the 1940s by <a href="/wiki/Albert_Coons" title="Albert Coons">Albert H. Coons</a>.<sup id="cite_ref-:2_2-1" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup><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><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><figure class="mw-halign-right" typeof="mw:File/Thumb"><a href="/wiki/File:Lupus_band_test.jpg" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/7/71/Lupus_band_test.jpg/280px-Lupus_band_test.jpg" decoding="async" width="280" height="210" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/7/71/Lupus_band_test.jpg/420px-Lupus_band_test.jpg 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/7/71/Lupus_band_test.jpg/560px-Lupus_band_test.jpg 2x" data-file-width="700" data-file-height="525" /></a><figcaption>Photomicrograph of a <a href="/wiki/Histology" title="Histology">histological section</a> of human skin prepared for <b>direct immunofluorescence</b> using an anti-IgG antibody. The skin is from a patient with systemic <a href="/wiki/Lupus_erythematosus" title="Lupus erythematosus">lupus erythematosus</a> and shows IgG deposit at two different places: The first is a band-like deposit along the epidermal <a href="/wiki/Basement_membrane" title="Basement membrane">basement membrane</a> ("lupus band test" is positive). The second is within the nuclei of the <a href="/wiki/Epidermis_(skin)" class="mw-redirect" title="Epidermis (skin)">epidermal</a> cells (anti-nuclear antibodies).</figcaption></figure> <p>Immunofluorescence is employed in foundational scientific investigations and clinical diagnostic endeavors, showcasing its multifaceted utility across diverse substrates, including tissue sections, cultured <a href="/wiki/Cell_lines" class="mw-redirect" title="Cell lines">cell lines</a>, or individual cells. Its usage includes analysis of the distribution of <a href="/wiki/Protein" title="Protein">proteins</a>, <a href="/wiki/Glycan" title="Glycan">glycans</a>, small biological and non-biological molecules, and visualization of structures such as intermediate-sized filaments.<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> </p><p>If the topology of a cell membrane is undetermined, epitope insertion into proteins can be used in conjunction with immunofluorescence to determine structures within the cell membrane.<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> Immunofluorescence (IF) can also be used as a “semi-quantitative” method to gain insight into the levels and localization patterns of DNA methylation. IF can additionally be used in combination with other, non-antibody methods of fluorescent staining, e.g., the use of <a href="/wiki/DAPI" title="DAPI">DAPI</a> to label <a href="/wiki/DNA" title="DNA">DNA</a>.<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><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> </p><p>Examination of immunofluorescence specimens can be conducted utilizing various microscope configurations, including the <a href="/wiki/Epifluorescence_microscope" class="mw-redirect" title="Epifluorescence microscope">epifluorescence microscope</a>, <a href="/wiki/Confocal_microscope" class="mw-redirect" title="Confocal microscope">confocal microscope</a>, and widefield microscope.<sup id="cite_ref-:4_12-0" class="reference"><a href="#cite_note-:4-12"><span class="cite-bracket">[</span>12<span class="cite-bracket">]</span></a></sup> </p> <meta property="mw:PageProp/toc" /> <div class="mw-heading mw-heading2"><h2 id="Types">Types</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=1" title="Edit section: Types"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <figure class="mw-default-size" typeof="mw:File/Thumb"><a href="/wiki/File:Main_antinuclear_antibody_patterns_on_immunofluorescence.png" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/d/d9/Main_antinuclear_antibody_patterns_on_immunofluorescence.png/220px-Main_antinuclear_antibody_patterns_on_immunofluorescence.png" decoding="async" width="220" height="161" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/d/d9/Main_antinuclear_antibody_patterns_on_immunofluorescence.png/330px-Main_antinuclear_antibody_patterns_on_immunofluorescence.png 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/d/d9/Main_antinuclear_antibody_patterns_on_immunofluorescence.png/440px-Main_antinuclear_antibody_patterns_on_immunofluorescence.png 2x" data-file-width="814" data-file-height="596" /></a><figcaption>Main <a href="/wiki/Antinuclear_antibody" title="Antinuclear antibody">antinuclear antibody</a> patterns on immunofluorescence.<sup id="cite_ref-13" class="reference"><a href="#cite_note-13"><span class="cite-bracket">[</span>13<span class="cite-bracket">]</span></a></sup></figcaption></figure> <div class="mw-heading mw-heading3"><h3 id="Preparation_of_fluorescence">Preparation of fluorescence</h3><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=2" title="Edit section: Preparation of fluorescence"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <p>To perform immunofluorescence staining, a <a href="/wiki/Fluorophore" title="Fluorophore">fluorophore</a> must be conjugated (“tagged”) to an antibody. Staining procedures can be applied to both retained intracellular expressed antibodies, or to cell surface antigens on living cells. There are two general classes of immunofluorescence techniques: primary (direct) and secondary (indirect).<sup id="cite_ref-:0_1-1" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-:2_2-2" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup> The following descriptions will focus primarily on these classes in terms of conjugated antibodies.<sup id="cite_ref-:4_12-1" class="reference"><a href="#cite_note-:4-12"><span class="cite-bracket">[</span>12<span class="cite-bracket">]</span></a></sup> </p> <div class="mw-heading mw-heading3"><h3 id="Primary_(direct)"><span id="Primary_.28direct.29"></span>Primary (direct)</h3><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=3" title="Edit section: Primary (direct)"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <figure typeof="mw:File/Thumb"><a href="/wiki/File:Primary_Immunofluorescence.png" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/4/43/Primary_Immunofluorescence.png/255px-Primary_Immunofluorescence.png" decoding="async" width="255" height="270" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/4/43/Primary_Immunofluorescence.png/383px-Primary_Immunofluorescence.png 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/4/43/Primary_Immunofluorescence.png/510px-Primary_Immunofluorescence.png 2x" data-file-width="722" data-file-height="764" /></a><figcaption>Basic concept of <b>Primary Immunofluorescence</b>: An antibody with a conjugated fluorophore, that is specifically bound to an epitope on the target molecule. </figcaption></figure> <p>Primary (direct) immunofluorescence (DIF) uses a single antibody, conjugated to a <a href="/wiki/Fluorophore" title="Fluorophore">fluorophore</a>. The antibody recognizes the target molecule (antigen) and binds to a specific region, called the <a href="/wiki/Epitope" title="Epitope">epitope</a>. The attached fluorophore can be detected via fluorescent microscopy, which, depending on the type of fluorophore, will emit a specific wavelength of light once excited.<sup id="cite_ref-:0_1-2" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-14" class="reference"><a href="#cite_note-14"><span class="cite-bracket">[</span>14<span class="cite-bracket">]</span></a></sup> </p><p>The direct attachment of the fluorophore to the antibody reduces the number of steps in the sample preparation procedure, saving time and reducing non-specific background signal during analysis.<sup id="cite_ref-:4_12-2" class="reference"><a href="#cite_note-:4-12"><span class="cite-bracket">[</span>12<span class="cite-bracket">]</span></a></sup> This also limits the possibility of antibody cross-reactivity, and possible mistakes throughout the process. One disadvantage of DIF is the limited number of antibodies that can bind to the antigen. This limitation may reduce sensitivity to the technique. When the target protein is available in only small concentrations, a better approach would be secondary IF, which is considered to be more sensitive than DIF <sup id="cite_ref-:2_2-3" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-:4_12-3" class="reference"><a href="#cite_note-:4-12"><span class="cite-bracket">[</span>12<span class="cite-bracket">]</span></a></sup> when compared to Secondary (Indirect) Immunofluorescence.<sup id="cite_ref-:0_1-3" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> </p> <figure typeof="mw:File/Thumb"><a href="/wiki/File:Secondary_Immunofluorescence.png" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/d/dd/Secondary_Immunofluorescence.png/254px-Secondary_Immunofluorescence.png" decoding="async" width="254" height="354" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/d/dd/Secondary_Immunofluorescence.png/381px-Secondary_Immunofluorescence.png 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/d/dd/Secondary_Immunofluorescence.png/508px-Secondary_Immunofluorescence.png 2x" data-file-width="786" data-file-height="1096" /></a><figcaption>Basic concept of <b>Secondary Immunofluorescence</b>: Secondary antibody, with a conjugated fluorophore, bound to a primary antibody that is specifically bound to an epitope on the target molecule.</figcaption></figure> <div class="mw-heading mw-heading3"><h3 id="Secondary_(indirect)"><span id="Secondary_.28indirect.29"></span>Secondary (indirect)</h3><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=4" title="Edit section: Secondary (indirect)"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <p>Secondary (indirect) immunofluorescence (SIF) is similar to direct immunofluorescence, however the technique utilizes two types of antibodies whereas only one of them have a conjugated fluorophore. The antibody with the conjugated fluorophore is referred to as the secondary antibody, while the unconjugated is referred to as the primary antibody.<sup id="cite_ref-:0_1-4" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> </p><p>The principle of this technique is that the primary antibody specifically binds to the epitope on the target molecule, whereas the secondary antibody, with the conjugated fluorophore, recognizes and binds to the primary antibody.<sup id="cite_ref-:0_1-5" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup>   </p><p>This technique is considered to be more sensitive than primary immunofluorescence, because multiple secondary antibodies can bind to the same primary antibody. The increased number of fluorophore molecules per antigen increases the amount of emitted light, and thus amplifies the signal.<sup id="cite_ref-:0_1-6" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> There are different methods for attaining a higher fluorophore-antigen ratio such as the Avidin-Biotin Complex (ABC method) and Labeled Streptavidin-Biotin (LSAB method).<sup id="cite_ref-15" class="reference"><a href="#cite_note-15"><span class="cite-bracket">[</span>15<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-16" class="reference"><a href="#cite_note-16"><span class="cite-bracket">[</span>16<span class="cite-bracket">]</span></a></sup> </p> <div class="mw-heading mw-heading2"><h2 id="Limitations">Limitations</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=5" title="Edit section: Limitations"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <figure typeof="mw:File/Thumb"><a href="/wiki/File:ABC-method.png" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/9/9e/ABC-method.png/257px-ABC-method.png" decoding="async" width="257" height="200" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/9/9e/ABC-method.png/386px-ABC-method.png 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/9/9e/ABC-method.png/514px-ABC-method.png 2x" data-file-width="557" data-file-height="434" /></a><figcaption>Basic concept of the <b>ABC-method</b>: Primary antibody binds to the antigen, before binding to the biotinylated secondary antibody. Avidin-Biotin enzyme complex (ABC) then attaches to the secondary antibody. </figcaption></figure> <p>Immunofluorescence is only limited to fixed (i.e. dead) cells, when studying structures within the cell, as antibodies generally do not penetrate intact cellular or subcellular membranes in living cells, because they are large proteins. To visualize these structures, antigenic material must be fixed firmly on its natural localization inside the cell.<sup id="cite_ref-17" class="reference"><a href="#cite_note-17"><span class="cite-bracket">[</span>17<span class="cite-bracket">]</span></a></sup> To study structures within living cells, in combination with fluorescence, one can utilize <a href="/wiki/Recombinant_protein" class="mw-redirect" title="Recombinant protein">recombinant proteins</a> containing fluorescent protein domains, e.g., <a href="/wiki/Green_fluorescent_protein" title="Green fluorescent protein">green fluorescent protein</a> (GFP). The GFP-technique involves altering the genetic information of the cells.<sup id="cite_ref-18" class="reference"><a href="#cite_note-18"><span class="cite-bracket">[</span>18<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-19" class="reference"><a href="#cite_note-19"><span class="cite-bracket">[</span>19<span class="cite-bracket">]</span></a></sup> </p><p>A significant problem with immunofluorescence is <a href="/wiki/Photobleaching" title="Photobleaching">photobleaching</a>,<sup id="cite_ref-:4_12-4" class="reference"><a href="#cite_note-:4-12"><span class="cite-bracket">[</span>12<span class="cite-bracket">]</span></a></sup> the fluorophores permanent loss of ability to emit light.<sup id="cite_ref-:0_1-7" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> To mitigate the risk of photobleaching one can employ different strategies. By reducing or limiting the intensity, or timespan of light exposure, the absorption-emission cycle of fluorescent light is decreased, thus preserving the fluorophores functionality. One can also increase the concentration of fluorophores, or opt for more robust fluorophores that exhibit resilience against photobleaching such as <a href="/wiki/Alexa_Fluor" title="Alexa Fluor">Alexa Fluors</a>, Seta Fluors, or <a href="/wiki/DyLight_Fluor" title="DyLight Fluor">DyLight Fluors</a>.<sup id="cite_ref-:2_2-4" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup> </p> <figure typeof="mw:File/Thumb"><a href="/wiki/File:LSAB-method.jpg" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/4/44/LSAB-method.jpg/259px-LSAB-method.jpg" decoding="async" width="259" height="199" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/4/44/LSAB-method.jpg/389px-LSAB-method.jpg 1.5x, //upload.wikimedia.org/wikipedia/commons/4/44/LSAB-method.jpg 2x" data-file-width="489" data-file-height="375" /></a><figcaption>Basic concept of the <b>LSAB-method</b>: Utilizes a Streptavidin–enzyme conjugate for the identification of the biotinylated secondary antibody which is bound to the primary antibody. This approach is applicable when the Avidin–Biotin complex in the ABC method becomes too large.</figcaption></figure> <p>Other problems that may arise when using immunofluorescence techniques include <a href="/wiki/Autofluorescence" title="Autofluorescence">autofluorescence</a>, spectral overlap and non-specific staining.<sup id="cite_ref-:0_1-8" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-:2_2-5" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup> Autofluorescence includes the natural fluorescence emitted from the sample tissue or cell itself. Spectral overlap happens when a fluorophore has a broad emission specter, that overlaps with the specter of another fluorophore, thus giving rise to false signals. Non-specific staining occurs when the antibody, containing the fluorophore, binds to unintended proteins because of sufficient similarity in the epitope. This can lead to false positives.<sup id="cite_ref-:2_2-6" class="reference"><a href="#cite_note-:2-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-:3_4-1" class="reference"><a href="#cite_note-:3-4"><span class="cite-bracket">[</span>4<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-:0_1-9" class="reference"><a href="#cite_note-:0-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> </p> <div class="mw-heading mw-heading2"><h2 id="Advances">Advances</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=6" title="Edit section: Advances"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <p>The main improvements to immunofluorescence lie in the development of fluorophores and fluorescent microscopes. Fluorophores can be structurally modified to improve brightness and photostability, while preserving spectral properties and cell permeability.<sup id="cite_ref-20" class="reference"><a href="#cite_note-20"><span class="cite-bracket">[</span>20<span class="cite-bracket">]</span></a></sup> </p> <figure typeof="mw:File/Thumb"><a href="/wiki/File:HSP_IF_IgA.jpg" class="mw-file-description"><img src="//upload.wikimedia.org/wikipedia/commons/thumb/1/1f/HSP_IF_IgA.jpg/286px-HSP_IF_IgA.jpg" decoding="async" width="286" height="215" class="mw-file-element" srcset="//upload.wikimedia.org/wikipedia/commons/thumb/1/1f/HSP_IF_IgA.jpg/429px-HSP_IF_IgA.jpg 1.5x, //upload.wikimedia.org/wikipedia/commons/thumb/1/1f/HSP_IF_IgA.jpg/572px-HSP_IF_IgA.jpg 2x" data-file-width="600" data-file-height="450" /></a><figcaption>Photomicrograph of a <a href="/wiki/Histology" title="Histology">histological section</a> of human skin prepared for <b>direct immunofluorescence</b> using an anti-IgA antibody. The skin is from a patient with <a href="/wiki/Henoch%E2%80%93Sch%C3%B6nlein_purpura" title="Henoch–Schönlein purpura">Henoch–Schönlein purpura</a>: IgA deposits are found in the walls of small superficial capillaries (yellow arrows). The pale wavy green area on top is the <a href="/wiki/Epidermis_(skin)" class="mw-redirect" title="Epidermis (skin)">epidermis</a>, the bottom fibrous area is the <a href="/wiki/Dermis" title="Dermis">dermis</a>.</figcaption></figure> <p><a href="/wiki/Super-resolution_microscopy" title="Super-resolution microscopy">Super-resolution fluorescence microscopy</a> methods can produce images with a higher resolution than those microscopes imposed by the <a href="/wiki/Diffraction-limited_system" title="Diffraction-limited system">diffraction limit</a>. This enables the determination of structural details within the cell.<sup id="cite_ref-:1_21-0" class="reference"><a href="#cite_note-:1-21"><span class="cite-bracket">[</span>21<span class="cite-bracket">]</span></a></sup> Super-resolution in fluorescence, more specifically, refers to the ability of a microscope to prevent the simultaneous fluorescence of adjacent spectrally identical fluorophores (spectral overlap). Some of the recently developed super-resolution fluorescent microscope methods include stimulated emission depletion (<a href="/wiki/STED_microscopy" title="STED microscopy">STED</a>) microscopy, saturated structured-illumination microscopy (SSIM), fluorescence photoactivation localization microscopy (F<a href="/wiki/Photoactivated_localization_microscopy" title="Photoactivated localization microscopy">PALM</a>), and stochastic optical reconstruction microscopy (STORM).<sup id="cite_ref-22" class="reference"><a href="#cite_note-22"><span class="cite-bracket">[</span>22<span class="cite-bracket">]</span></a></sup> </p> <div class="mw-heading mw-heading2"><h2 id="Notable_people">Notable people</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=7" title="Edit section: Notable people"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <ul><li><a href="/wiki/Albert_Coons" title="Albert Coons">Albert Hewett Coons</a> (1912–1978), <a href="/wiki/Physician" title="Physician">physician</a>, <a href="/wiki/Pathologist" class="mw-redirect" title="Pathologist">pathologist</a> and <a href="/wiki/Immunologist" class="mw-redirect" title="Immunologist">immunologist</a></li> <li><a href="/wiki/Cornelia_Mitchell_Downs" title="Cornelia Mitchell Downs">Cornelia Mitchell Downs</a> (1892–1987), <a href="/wiki/Microbiologist" title="Microbiologist">microbiologist</a> and <a href="/wiki/Journalist" title="Journalist">journalist</a></li></ul> <div class="mw-heading mw-heading2"><h2 id="See_also">See also</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=8" title="Edit section: See also"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <ul><li><a href="/wiki/Antibody" title="Antibody">Antibodies</a></li> <li><a href="/wiki/Cutaneous_conditions_with_immunofluorescence_findings" class="mw-redirect" title="Cutaneous conditions with immunofluorescence findings">Cutaneous conditions with immunofluorescence findings</a></li> <li><a href="/wiki/Fluorescence" title="Fluorescence">Fluorescence</a></li> <li><a href="/wiki/Immunochemistry" title="Immunochemistry">Immunochemistry</a></li> <li><a href="/wiki/Immunohistochemistry" title="Immunohistochemistry">Immunohistochemistry</a></li> <li><a href="/wiki/Patching_and_Capping" title="Patching and Capping">Patching and Capping</a></li></ul> <div class="mw-heading mw-heading2"><h2 id="References">References</h2><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="/w/index.php?title=Immunofluorescence&action=edit&section=9" title="Edit section: References"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <style data-mw-deduplicate="TemplateStyles:r1239543626">.mw-parser-output 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href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835776">2835776</a></span>. <a href="/wiki/PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/19489737">19489737</a>.</cite><span title="ctx_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.jtitle=Annual+Review+of+Biochemistry&rft.atitle=Super-resolution+fluorescence+microscopy&rft.volume=78&rft.pages=%3Cspan+class%3D%22nowrap%22%3E993-%3C%2Fspan%3E1016&rft.date=2009-06-02&rft_id=https%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fpmc%2Farticles%2FPMC2835776%23id-name%3DPMC&rft_id=info%3Apmid%2F19489737&rft_id=info%3Adoi%2F10.1146%2Fannurev.biochem.77.061906.092014&rft.aulast=Huang&rft.aufirst=B&rft.au=Bates%2C+M&rft.au=Zhuang%2C+X&rft_id=https%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fpmc%2Farticles%2FPMC2835776&rfr_id=info%3Asid%2Fen.wikipedia.org%3AImmunofluorescence" class="Z3988"></span></span> </li> <li 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External links"><span>edit</span></a><span class="mw-editsection-bracket">]</span></span></div> <ul><li><a rel="nofollow" class="external text" href="http://www.ii.bham.ac.uk/clinicalimmunology/CISimagelibrary/">Images associated with autoimmune diseases</a> <a rel="nofollow" class="external text" href="https://web.archive.org/web/20090925190032/http://www.ii.bham.ac.uk/clinicalimmunology/CISimagelibrary/">Archived</a> 2009-09-25 at the <a href="/wiki/Wayback_Machine" title="Wayback Machine">Wayback Machine</a> at <a href="/wiki/University_of_Birmingham" title="University of Birmingham">University of Birmingham</a></li> <li><a rel="nofollow" class="external text" href="http://www.bio.davidson.edu/COURSES/genomics/method/IMF.html">Overview</a> at <a href="/wiki/Davidson_College" title="Davidson College">Davidson College</a></li> <li><a rel="nofollow" class="external text" href="https://meshb.nlm.nih.gov/record/ui?name=Immunofluorescence">Immunofluorescence</a> at the U.S. National 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pathology</th><td class="navbox-list-with-group navbox-list navbox-odd hlist" style="width:100%;padding:0"><div style="padding:0 0.25em"> <ul><li><a href="/wiki/Disease" title="Disease">Disease</a> <ul><li><a href="/wiki/Infection" title="Infection">Infection</a></li> <li><a href="/wiki/Neoplasm" title="Neoplasm">Neoplasia</a></li> <li><a href="/wiki/Cause_(medicine)" title="Cause (medicine)">Cause</a></li> <li><a href="/wiki/Pathogenesis" title="Pathogenesis">Pathogenesis</a></li></ul></li> <li><a href="/wiki/Hemodynamics" title="Hemodynamics">Hemodynamics</a> <ul><li><a href="/wiki/Ischemia" title="Ischemia">Ischemia</a></li></ul></li> <li><a href="/wiki/Inflammation" title="Inflammation">Inflammation</a></li> <li><a href="/wiki/Cell_damage" title="Cell damage">Cell damage</a></li> <li><a href="/wiki/Wound_healing" title="Wound healing">Wound healing</a></li></ul> <dl><dt><a href="/wiki/Cellular_adaptation" title="Cellular adaptation">Cellular adaptation</a></dt> <dd></dd> <dd><a href="/wiki/Atrophy" title="Atrophy">Atrophy</a></dd> <dd><a href="/wiki/Hypertrophy" title="Hypertrophy">Hypertrophy</a></dd> <dd><a href="/wiki/Hyperplasia" title="Hyperplasia">Hyperplasia</a></dd> <dd><a href="/wiki/Dysplasia" title="Dysplasia">Dysplasia</a></dd> <dd><a href="/wiki/Metaplasia" title="Metaplasia">Metaplasia</a> <dl><dd><a href="/wiki/Squamous_metaplasia" title="Squamous metaplasia">Squamous</a></dd> <dd><a href="/wiki/Glandular_metaplasia" title="Glandular metaplasia">Glandular</a></dd></dl></dd></dl> <dl><dt><a href="/wiki/Cell_death" title="Cell death">Cell death</a></dt> <dd></dd> <dd><a href="/wiki/Necrosis" title="Necrosis">Necrosis</a> <dl><dd><a href="/wiki/Coagulative_necrosis" title="Coagulative necrosis">Coagulative necrosis</a></dd> <dd><a href="/wiki/Liquefactive_necrosis" title="Liquefactive necrosis">Liquefactive necrosis</a></dd> <dd><a href="/wiki/Gangrene" title="Gangrene">Gangrenous necrosis</a></dd> <dd><a href="/wiki/Caseous_necrosis" title="Caseous necrosis">Caseous necrosis</a></dd> <dd><a href="/wiki/Fat_necrosis" title="Fat necrosis">Fat necrosis</a></dd> <dd><a href="/wiki/Fibrinoid_necrosis" title="Fibrinoid necrosis">Fibrinoid necrosis</a></dd> <dd><a href="/wiki/Myocytolysis" title="Myocytolysis">Myocytolysis</a></dd></dl></dd> <dd><a href="/wiki/Programmed_cell_death" title="Programmed cell death">Programmed cell death</a> <dl><dd><a href="/wiki/Apoptosis" title="Apoptosis">Apoptosis</a></dd></dl></dd> <dd><a href="/wiki/Pyknosis" title="Pyknosis">Pyknosis</a></dd> <dd><a href="/wiki/Karyorrhexis" title="Karyorrhexis">Karyorrhexis</a></dd> <dd><a href="/wiki/Karyolysis" title="Karyolysis">Karyolysis</a></dd></dl> <dl><dt><i>Accumulations</i></dt> <dd><i><a href="/wiki/Pigment" title="Pigment">pigment</a></i> <dl><dd><a href="/wiki/Hemosiderin" title="Hemosiderin">Hemosiderin</a></dd> <dd><a href="/wiki/Lipochrome" class="mw-redirect" title="Lipochrome">Lipochrome</a>/<a href="/wiki/Lipofuscin" title="Lipofuscin">Lipofuscin</a></dd> <dd><a href="/wiki/Melanin" title="Melanin">Melanin</a></dd></dl></dd> <dd><a href="/wiki/Steatosis" title="Steatosis">Steatosis</a></dd></dl> </div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="/wiki/Anatomical_pathology" title="Anatomical pathology">Anatomical pathology</a></th><td class="navbox-list-with-group navbox-list navbox-even hlist" style="width:100%;padding:0"><div style="padding:0 0.25em"> <ul><li><a href="/wiki/Surgical_pathology" title="Surgical pathology">Surgical pathology</a></li> <li><a href="/wiki/Cytopathology" title="Cytopathology">Cytopathology</a></li> <li><a href="/wiki/Autopsy" title="Autopsy">Autopsy</a></li> <li><a href="/wiki/Molecular_pathology" title="Molecular pathology">Molecular pathology</a></li> <li><a href="/wiki/Forensic_pathology" title="Forensic pathology">Forensic pathology</a></li> <li><a href="/wiki/Oral_and_maxillofacial_pathology" title="Oral and maxillofacial pathology">Oral and maxillofacial pathology</a></li></ul> <ul><li><a href="/wiki/Gross_processing" title="Gross processing">Gross processing</a></li> <li><a href="/wiki/Histopathology" title="Histopathology">Histopathology</a></li> <li><a href="/wiki/Immunohistochemistry" title="Immunohistochemistry">Immunohistochemistry</a></li> <li><a href="/wiki/Electron_microscope" title="Electron microscope">Electron microscopy</a></li> <li><a class="mw-selflink selflink">Immunofluorescence</a></li> <li><a href="/wiki/Fluorescence_in_situ_hybridization" title="Fluorescence in situ hybridization">Fluorescence in situ hybridization</a></li></ul> </div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="/wiki/Clinical_pathology" title="Clinical pathology">Clinical pathology</a></th><td class="navbox-list-with-group navbox-list navbox-odd hlist" style="width:100%;padding:0"><div style="padding:0 0.25em"> <ul><li><a href="/wiki/Clinical_chemistry" title="Clinical chemistry">Clinical chemistry</a></li> <li><a href="/wiki/Hematopathology" title="Hematopathology">Hematopathology</a></li> <li><a href="/wiki/Transfusion_medicine" title="Transfusion medicine">Transfusion medicine</a></li> <li><a href="/wiki/Medical_microbiology" title="Medical microbiology">Medical microbiology</a></li> <li><a href="/wiki/Immunoassay" title="Immunoassay">Diagnostic immunology</a></li> <li><a href="/wiki/Immunopathology" title="Immunopathology">Immunopathology</a></li></ul> <ul><li><a href="/wiki/Enzyme_assay" title="Enzyme assay">Enzyme assay</a></li> <li><a href="/wiki/Mass_spectrometry" title="Mass spectrometry">Mass spectrometry</a></li> <li><a href="/wiki/Chromatography" title="Chromatography">Chromatography</a></li> <li><a href="/wiki/Flow_cytometry" title="Flow cytometry">Flow cytometry</a></li> <li><a href="/wiki/Blood_bank" title="Blood bank">Blood bank</a></li> <li><a href="/wiki/Microbiological_culture" title="Microbiological culture">Microbiological culture</a></li> <li><a href="/wiki/Serology" title="Serology">Serology</a></li></ul> </div></td></tr></tbody></table></div> <div class="navbox-styles"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1129693374"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1236075235"></div><div role="navigation" class="navbox" aria-labelledby="Medical_tests_used_in_immunology_(CPT_86000–86849)179" style="padding:3px"><table class="nowraplinks hlist mw-collapsible autocollapse navbox-inner" style="border-spacing:0;background:transparent;color:inherit"><tbody><tr><th scope="col" class="navbox-title" colspan="2"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1129693374"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1239400231"><div class="navbar plainlinks hlist navbar-mini"><ul><li class="nv-view"><a href="/wiki/Template:Immunologic_techniques_and_tests" title="Template:Immunologic techniques and 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class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em"> <ul><li><a href="/wiki/Chromatin_immunoprecipitation" title="Chromatin immunoprecipitation">Chromatin immunoprecipitation</a></li> <li><a href="/wiki/Immunodiffusion" title="Immunodiffusion">Immunodiffusion</a> <ul><li><a href="/wiki/Ouchterlony_double_immunodiffusion" title="Ouchterlony double immunodiffusion">Ouchterlony double immunodiffusion</a></li> <li><a href="/wiki/Radial_immunodiffusion" title="Radial immunodiffusion">Radial immunodiffusion</a></li> <li><a href="/wiki/Immunoelectrophoresis" title="Immunoelectrophoresis">Immunoelectrophoresis</a></li> <li><a href="/wiki/Counterimmunoelectrophoresis" title="Counterimmunoelectrophoresis">Counterimmunoelectrophoresis</a></li></ul></li></ul> </div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="/wiki/Immunoassay" title="Immunoassay">Immunoassay</a></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="/wiki/Chemiluminescent_immunoassay" title="Chemiluminescent immunoassay">Chemiluminescent immunoassay</a></li> <li><a href="/wiki/ELISA" title="ELISA">ELISA</a></li> <li><a href="/wiki/ELISpot" title="ELISpot">ELISpot</a></li> <li><a href="/wiki/Enzyme_multiplied_immunoassay_technique" title="Enzyme multiplied immunoassay technique">EMIT</a></li> <li><a href="/wiki/Radioallergosorbent_test" title="Radioallergosorbent test">RAST test</a></li> <li><a href="/wiki/Radioimmunoassay" title="Radioimmunoassay">Radioimmunoassay</a></li> <li><a href="/wiki/Radiobinding_assay" title="Radiobinding assay">Radiobinding assay</a></li> <li><a class="mw-selflink selflink">Immunofluorescence</a></li> <li><a href="/wiki/Turbidimetric_inhibition_immunoassay" title="Turbidimetric inhibition immunoassay">TINIA</a></li> <li><a href="/wiki/Fluorescence_polarization_immunoassay" title="Fluorescence polarization immunoassay">FPIA</a></li> <li><a href="/wiki/Cloned_enzyme_donor_immunoassay" title="Cloned enzyme donor immunoassay">CEDIA</a></li> <li><a href="/wiki/Surround_optical-fiber_immunoassay" title="Surround optical-fiber immunoassay">SOFIA</a></li> <li><a href="/wiki/Lateral_flow_test" title="Lateral flow test">Rapid test</a></li> <li><a href="/wiki/Surface_plasmon_resonance#SPR_immunoassay" title="Surface plasmon resonance">SPR_IA</a></li> <li><a href="/wiki/Surface-enhanced_Raman_spectroscopy#Applications" title="Surface-enhanced Raman spectroscopy">SERS_IA</a></li></ul> </div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="/wiki/Agglutination_(biology)" title="Agglutination (biology)">Agglutination</a></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="/wiki/Hemagglutination" title="Hemagglutination">Hemagglutination</a>/<a href="/wiki/Hemagglutinin" title="Hemagglutinin">Hemagglutinin</a> <ul><li><a href="/wiki/Coombs_test" title="Coombs test">Coombs test</a></li></ul></li> <li><a href="/wiki/Latex_fixation_test" title="Latex fixation test">Latex fixation test</a></li></ul> </div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Other</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="/wiki/Diagnostic_immunology" class="mw-redirect" title="Diagnostic immunology">Diagnostic immunology</a></li> <li><a href="/wiki/Nephelometry" class="mw-redirect" title="Nephelometry">Nephelometry</a></li> <li><a href="/wiki/Complement_fixation_test" title="Complement fixation test">Complement fixation test</a></li> <li><a href="/wiki/Immunocytochemistry" title="Immunocytochemistry">Immunocytochemistry</a></li> <li><a href="/wiki/Immunohistochemistry" title="Immunohistochemistry">Immunohistochemistry</a> <ul><li><a href="/wiki/Direct_fluorescent_antibody" title="Direct fluorescent antibody">Direct fluorescent antibody</a></li></ul></li> <li><a href="/wiki/Epitope_mapping" title="Epitope mapping">Epitope mapping</a></li> <li><a href="/wiki/Skin_allergy_test" title="Skin allergy test">Skin allergy test</a></li> <li><a href="/wiki/Patch_test" title="Patch test">Patch test</a></li> <li><a href="/wiki/Total_complement_activity" title="Total complement activity">Total complement activity</a></li> <li><a href="/wiki/MELISA" title="MELISA">MELISA</a></li></ul> </div></td></tr></tbody></table></div> <div class="navbox-styles"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1129693374"><link rel="mw-deduplicated-inline-style" href="mw-data:TemplateStyles:r1236075235"><style data-mw-deduplicate="TemplateStyles:r1038841319">.mw-parser-output .tooltip-dotted{border-bottom:1px dotted;cursor:help}</style><link rel="mw-deduplicated-inline-style" 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