{"id":24855,"date":"2022-03-13T03:44:00","date_gmt":"2022-03-12T21:59:00","guid":{"rendered":"https:\/\/microbenotes.com\/?p=24855"},"modified":"2022-05-04T02:42:08","modified_gmt":"2022-05-03T20:57:08","slug":"iron-hematoxylin-staining","status":"publish","type":"post","link":"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/","title":{"rendered":"Iron-Hematoxylin Staining"},"content":{"rendered":"<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_65 counter-hierarchy ez-toc-counter ez-toc-custom ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<p class=\"ez-toc-title \" >Table of Contents<\/p>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #0a0a0a;color:#0a0a0a\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #0a0a0a;color:#0a0a0a\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#what-is-iron-hematoxylin-staining\" title=\"What is Iron-Hematoxylin Staining?\">What is Iron-Hematoxylin Staining?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#principle-of-iron-hematoxylin-staining\" title=\"Principle of Iron-Hematoxylin Staining\">Principle of Iron-Hematoxylin Staining<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#objectives-of-iron-hematoxylin-staining\" title=\"Objectives of Iron-Hematoxylin Staining\">Objectives of Iron-Hematoxylin Staining<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#reagents\" title=\"Reagents\">Reagents<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#procedure-of-iron-hematoxylin-staining\" title=\"Procedure of Iron-Hematoxylin Staining\">Procedure of Iron-Hematoxylin Staining<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-6\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#preparation-of-solutions\" title=\"Preparation of solutions\">Preparation of solutions<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-7\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#preparation-of-pva-samples\" title=\"Preparation of PVA-samples\">Preparation of PVA-samples<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-8\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#preparation-of-saf-samples\" title=\"Preparation of SAF-samples\">Preparation of SAF-samples<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-9\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#staining-procedure\" title=\"Staining Procedure\">Staining Procedure<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-10\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#a-fresh-samples\" title=\"a. Fresh samples:\">a. Fresh samples:<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-11\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#b-pva-preserves-samples\" title=\"b. PVA-Preserves samples:\">b. PVA-Preserves samples:<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-12\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#c-staining-saf-preserved-samples\" title=\"c. Staining SAF preserved samples\">c. Staining SAF preserved samples<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-13\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#result-and-interpretation-of-iron-hematoxylin-staining\" title=\"Result and Interpretation of Iron-Hematoxylin Staining\">Result and Interpretation of Iron-Hematoxylin Staining<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-14\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#applications-of-iron-hematoxylin-staining\" title=\"Applications of Iron-Hematoxylin Staining\">Applications of Iron-Hematoxylin Staining<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-15\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#advantages-of-iron-hematoxylin-staining\" title=\"Advantages of Iron-Hematoxylin Staining\">Advantages of Iron-Hematoxylin Staining<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-16\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#limitations-of-iron-hematoxylin-staining\" title=\"Limitations of Iron-Hematoxylin Staining\">Limitations of Iron-Hematoxylin Staining<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-17\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/#references-and-sources\" title=\"References and Sources\">References and Sources<\/a><\/li><\/ul><\/nav><\/div>\n<h2><span class=\"ez-toc-section\" id=\"what-is-iron-hematoxylin-staining\"><\/span><strong>What is Iron-Hematoxylin Staining?<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>This is a traditional staining technique commonly used for the detection of intestinal protozoans majorly because of its permanence and ability to demonstrate, observe, and quantify with high clarity protozoans nuclear structures. <b>Iron hematoxylin<\/b>\u00a0was the\u00a0<b>stain<\/b> used for most of the original morphological descriptions of intestinal <strong>protozoa<\/strong> found in humans.<\/p>\n<p>The iron hematoxylin staining method can stain the intracellular and nuclear structure&#8217;s dark compared to the cytoplasm, for easy identification of parasite morphologies.<\/p>\n<p>Iron hematoxylin staining methods can be used with either fresh specimen, or sodium acetate acetic acid formalin (SAF)-preserved, or <em>polyvinyl-alcohol (<\/em>PVA)-preserved specimens.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"principle-of-iron-hematoxylin-staining\"><\/span><strong>Principle of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<div class=\"t m0 x7 h7 yc ff2 fs3 fc0 sc0 lse wsc\">Hematoxylin is a natural dye that is extracted from <em><span class=\"ff5 ls10 wse\">Haematoxylon campechianum<\/span><\/em><span class=\"ls11 wsf\">, a leguminous plant, which must undergo oxidation (Ripening) to form hematein. Oxidation is a slow process involving dissolving crystals of Hematoxylin in water and its then exposed to oxygen (Atmospheric) and the addition of hydrogen peroxide with close monitoring to avoid overoxidation. Over-oxidation leads to the formation of oxy-hematein which is not a dye. <\/span><\/div>\n<p><span class=\"ls11 wsf\">A hematoxylin solution is made up of a combination of hematoxylin, hematein, oxihematein.\u00a0 Hematein acts as the mordant to produce a lake i.e it <\/span>reacts with ferric ammonium sulfate to produce the ferric lake (iron-hematoxylin), a basic dye. The iron-hematoxylin compound is stable and stains sharply the structures used to identify intestinal protozoan.<\/p>\n<div>It is mostly used to stain regressively, i.e the slides are first overstained and then differentiated. The use of Alcoholic iodine ensures complete fixation of the smear.<\/div>\n<h2><span class=\"ez-toc-section\" id=\"objectives-of-iron-hematoxylin-staining\"><\/span><strong>Objectives of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ul>\n<li>To demonstrate the presence of intestinal parasites from a given stool sample.<\/li>\n<li>To differentiate protozoal parasite morphologies.<\/li>\n<\/ul>\n<h3><span class=\"ez-toc-section\" id=\"reagents\"><\/span><strong>Reagents<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<div>Distilled water, Mercuric Chloride, Ethyl alcohol, Iodine, Ferric Ammonium Sulfate (Iron Alum), Hydrochloric acid, Hematoxylin, alcoholic iodine<\/div>\n<h2><span class=\"ez-toc-section\" id=\"procedure-of-iron-hematoxylin-staining\"><\/span><strong>Procedure of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<h3><span class=\"ez-toc-section\" id=\"preparation-of-solutions\"><\/span><strong>Preparation of solutions<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li><strong>Schaudinn\u2019s solution:<\/strong> Into 80ml of distilled water, add a saturated solution of Mercuric Chloride, Ethyl alcohol 20ml (95%), Acetic acid 3 mL<\/li>\n<li><strong>Iodine solution:<\/strong> add 98ml of Ethyl alcohol (95%) and 2ml of Tincture of iodine (2%)<\/li>\n<li><strong>The mordant-differentiating solution:<\/strong> Iron alum (ferric ammonium sulfate) 2 g, 1ml of Hydrochloric acid, distilled water 98 mL<\/li>\n<li><strong>Hematoxylin solution:<\/strong> Hematoxylin 0.30 g, distilled water 100 mL<\/li>\n<\/ol>\n<p><strong>Note<\/strong>: Iron Hematoxylin Stain and Mordant-differentiating solutions are stable individually for 12 months when stored at the right temperature. Once the working solution is prepared, let it remains stable for 7 days before using it for staining. Filtering of the stains is recommended periodically using a filter paper to remove any debris that may be present and always keep seal the container to avoid evaporation.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"preparation-of-pva-samples\"><\/span><strong>Preparation of PVA-samples<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li>PVA-preserved samples should be allowed to fix for at least 30 min and mix the content with two applicator sticks.<\/li>\n<li>Pour some of the above PVA mixtures to a paper towel and allow to stand for 2-3 minutes for effective absorption of PVA.<\/li>\n<li>Apply some of the stool samples on the paper towel to the glass slides using an applicator stick and allow it to dry in an incubator at 37\u00b0C or overnight at room temperature to ensure the smear is completely dry.<\/li>\n<li>To the dry smear, add iodine-alcohol.<\/li>\n<\/ol>\n<h3><span class=\"ez-toc-section\" id=\"preparation-of-saf-samples\"><\/span><strong>Preparation of SAF-samples<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li>A SAF-stool sample is mixed and strained through a wire gauze to a 15ml centrifuge tube and centrifuge for 1 minute at 500xg<\/li>\n<li>Decant the supernatant and centrifuged for 10min at 500xg and collect sediments of 0.5ml to 10ml. If necessary, adjust by repeating step 1 or by resuspending the sediment in saline (0.85% NaCl) and removing part of the suspension.<\/li>\n<li>Prepare a smear on a clean glass slide, from the sediments for later, by adding a drop of Mayer&#8217;s albumin on the smear. Then add a drop of the SAF-preserved sample sediments and allow them to dry at room temperature for 30 minutes before staining.<\/li>\n<li>You can (NOT MUST) also postfix with Schaudinn&#8217;s solution before staining before initiating a trichrome stain procedure with 70% alcohol rinse then adding iodine- alcohol.<\/li>\n<li>After drying, place the smear in 70% alcohol<\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"staining-procedure\"><\/span><strong>Staining Procedure<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<h3><span class=\"ez-toc-section\" id=\"a-fresh-samples\"><\/span><strong>a. Fresh samples:<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li>On clean microscopic glass slides, slides, prepare thin smears of the stool samples using an applicator stick and while still wet, dip the slides into Schaudinn\u2019s solution, for at least 30 minutes. This is called the Schaudinn&#8217;s fixation<\/li>\n<li>Place the Schaudinn&#8217;s fixed smear in 70% alcohol to remove excess fixative.<\/li>\n<li>Rinse the slide in smoothly flowing tap water at least three times<\/li>\n<li>Place slide in iron hematoxylin working solution for 4 to 5 min<\/li>\n<li>Wash slides with smooth running tap water (constant stream of water into the container) for 10 min.<\/li>\n<li>Immerse the slides in 95% ethyl alcohol for 5 minutes.<\/li>\n<li>Place the slides in 100% ethanol for 5 minutes.<\/li>\n<li>Immerse the slides in two changes in xylene for 5 minutes for each change<\/li>\n<li>Add a permount (Mounting Media is formulated for mounting and storing long-term slides) to the stain and cover with a coverslip.<\/li>\n<li>Examine the smear under a microscope at 100x and also under oil immersion before reporting.<\/li>\n<\/ol>\n<h3><span class=\"ez-toc-section\" id=\"b-pva-preserves-samples\"><\/span><strong>b. PVA-Preserves samples:<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li>On clean microscopic glass slides, slides, prepare thin smears of the stool samples using an applicator stick and while still wet, dip the slides into Schaudinn\u2019s solution, for at least 30 minutes. This is called the Schaudinn&#8217;s fixation<\/li>\n<li>If the fresh specimen is liquid, place 3 to 4 drops of PVA on the slide, mix several drops of fecal material, spread and allow to dry overnight at room temperature, or for a few hours in an incubator at 37oc.\u00a0 Note: this procedure is applied only for liquid samples for uniform mixing and not semi-solid or solid stool samples.<\/li>\n<li>Proceed with the trichrome staining procedure by placing the slides in iodine-alcohol.<\/li>\n<li>Place in 70% ethanol for 5 minutes and wash in smooth running tap water for 10 minutes<\/li>\n<li>Place slide in iron hematoxylin working solution for 4 to 5 min<\/li>\n<li>Wash slides with smooth running tap water for 10 min.<\/li>\n<li>Immerse the slides in 70% ethanol for 5 minutes.<\/li>\n<li>Then immerse the slides again in 95% ethanol for another 5 minutes<\/li>\n<li>In two changes, place the slides in 100% ethanol for 5 minutes for each change.<\/li>\n<li>Immerse the slides in two changes in xylene for 5 minutes for each change<\/li>\n<li>Add a permount (Mounting Media is formulated for mounting and storing long-term slides) to the stain and cover with a coverslip.<\/li>\n<li>Examine the smear under a microscope at 100x and also under oil immersion before reporting.<\/li>\n<\/ol>\n<h3><span class=\"ez-toc-section\" id=\"c-staining-saf-preserved-samples\"><\/span><strong>c. Staining SAF preserved samples<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ol>\n<li>Prepare slides for staining using the SChaudinn solution and let it air dry for a few hours in an incubator or overnight at room temperature.<\/li>\n<li>Place the slides in 70% ethanol for 5 min and fix with SAF as described above.<\/li>\n<li>Wash slide in smoothly running tap water for 10 minutes.<\/li>\n<li>Place the slides in iron hematoxylin working solution for 4 to 5 min.<\/li>\n<li>Wash the slides in smoothly running tap water for 10 minutes.<\/li>\n<li>Place the slides in 70% ethanol for 5 min.<\/li>\n<li>Then, place the slides in 95% ethanol for 5 min.<\/li>\n<li>Followed by immersing the slides in 100% ethanol in two changes for 5 minutes for each change.<\/li>\n<li>Immerse the slide in two changes of xylene for 5 minutes for each change.<\/li>\n<li>Add a permount (Mounting Media is formulated for mounting and storing long-term slides) to the stain and cover with a coverslip.<\/li>\n<li>Examine the smear under a microscope at 100x and also under oil immersion before reporting.<\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"result-and-interpretation-of-iron-hematoxylin-staining\"><\/span><strong>Result and Interpretation of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>Using the iron hematoxylin staining procedure, with a specimen containing microorganisms, the cytoplasm of trophozoites and cysts will stain violet. The nuclear chromatin, chromatoid bodies, erythrocytes, and bacteria stain dark purple to black.<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-30282\" src=\"https:\/\/microbenotes.com\/wp-content\/uploads\/2020\/02\/Iron-Hematoxylin-Staining.jpg\" alt=\"Iron-Hematoxylin Staining\" width=\"800\" height=\"420\" srcset=\"https:\/\/microbenotes.com\/wp-content\/uploads\/2020\/02\/Iron-Hematoxylin-Staining.jpg 800w, https:\/\/microbenotes.com\/wp-content\/uploads\/2020\/02\/Iron-Hematoxylin-Staining-300x158.jpg 300w, https:\/\/microbenotes.com\/wp-content\/uploads\/2020\/02\/Iron-Hematoxylin-Staining-768x403.jpg 768w\" sizes=\"(max-width: 800px) 100vw, 800px\" \/><\/p>\n<p>Figure: Photomicrographs of enteric protozoa stained with a modified iron-hematoxylin stain (incorporating a carbol fuschin staining step). (A) <em>Cyclospora<\/em> oocysts; (B) <em>Cryptosporidium<\/em> oocysts; (C) <em>Dientamoeba fragilis<\/em> binucleated trophozoite; (D) <em>Dientamoeba fragilis<\/em> uninucleated trophozoite; (E) <em>Entamoeba histolytica<\/em> cysts; (F) <em>Entamoeba histolytica<\/em> trophozoite; (G) <em>Giardia<\/em> cysts; (H) <em>Giardia<\/em> trophozoites. Image Source: <a href=\"https:\/\/cmr.asm.org\/content\/22\/4\/634\" target=\"_blank\" rel=\"noopener noreferrer\">American Society for Microbiology<\/a>.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"applications-of-iron-hematoxylin-staining\"><\/span><strong>Applications of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ul>\n<li>For diagnosis of parasitic infections by the demonstration of protozoal cysts and trophozoites in specimens.<\/li>\n<li>It prepares permanent stains which are important for reference.<\/li>\n<\/ul>\n<h3><span class=\"ez-toc-section\" id=\"advantages-of-iron-hematoxylin-staining\"><\/span><strong>Advantages of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ul>\n<li>It is adequate for routine use in the diagnosis of intestinal protozoans.<\/li>\n<li>It offers high clarity for the differentiation of protozoans.<\/li>\n<\/ul>\n<h3><span class=\"ez-toc-section\" id=\"limitations-of-iron-hematoxylin-staining\"><\/span><strong>Limitations of Iron-Hematoxylin Staining<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<ul>\n<li>If the fixation is not properly done, the result will be distorted.<\/li>\n<li>It is a slow procedure that requires adequate time to perform.<\/li>\n<li>Schaudinn solution contains mercury and therefore it must be removed completely before staining. it interferes with the staining reagents.<\/li>\n<\/ul>\n<h2><span class=\"ez-toc-section\" id=\"references-and-sources\"><\/span><strong>References and Sources<\/strong><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ul>\n<li><span style=\"font-size: 10pt;\">1% &#8211; https:\/\/www.researchgate.net\/publication\/26274071_Pancreatic_Remodeling_Beta-Cell_Apoptosis_Proliferation_and_Neogenesis_and_the_Measurement_of_Beta-Cell_Mass_and_of_Individual_Beta-Cell_Size<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">1% &#8211; https:\/\/www.researchgate.net\/post\/Does_anyone_has_done_a_very_good_luxol_fast_blue_staining_using_cryosections<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">1% &#8211; https:\/\/www.dalynn.com\/dyn\/ck_assets\/files\/tech\/SI70.pdf<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">1% &#8211; https:\/\/citeseerx.ist.psu.edu\/viewdoc\/download?doi=10.1.1.832.446&amp;rep=rep1&amp;type=pdf<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">1% &#8211; http:\/\/www.med-chem.com\/para-site.php?url=procedures&amp;subsection=microscopic&amp;tertiary_section=permanent_stained_smear&amp;quad_section=pss_iron_hematoxylin_stain<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/www.jove.com\/t\/3954\/rat-mesentery-exteriorization-model-for-investigating-cellular<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/www.cdc.gov\/meningitis\/lab-manual\/chpt06-culture-id.html<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/www.cdc.gov\/dpdx\/diagnosticprocedures\/stool\/staining.html<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/www.abcam.com\/ps\/pdf\/protocols\/ihc_p.pdf<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/science.howstuffworks.com\/engineering\/structural\/water-slide4.htm<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/microbeonline.com\/types-of-staining-techniques-used-in-microbiology-and-their-applications\/<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/eduwavepool.unizwa.edu.om\/lmsdatapool\/00011253\/LearningObjects\/Parasitology%20Practical%20Manual.doc<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; https:\/\/chem.libretexts.org\/Bookshelves\/Organic_Chemistry\/Book%3A_Basic_Principles_of_Organic_Chemistry_(Roberts_and_Caserio)\/22%3A_Arenes%2C_Electrophilic_Aromatic_Substitution\/22.10%3A_Oxidation_Reactions<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; http:\/\/www.atlas-protozoa.com\/microscope-exam-ed.php<\/span><\/li>\n<li><span style=\"font-size: 10pt;\">&lt;1% &#8211; http:\/\/shs-manual.ucsc.edu\/policy\/fecal-microscopic-examination<\/span><\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>What is Iron-Hematoxylin Staining? This is a traditional staining technique commonly used for the detection of intestinal protozoans majorly because of its permanence and ability to demonstrate, observe, and quantify &#8230; <a title=\"Iron-Hematoxylin Staining\" class=\"read-more\" href=\"https:\/\/microbenotes.com\/iron-hematoxylin-staining\/\" aria-label=\"Read more about Iron-Hematoxylin Staining\">Read more<\/a><\/p>\n","protected":false},"author":3,"featured_media":30282,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[705],"tags":[5599,5164,988],"custom":{"td_video":"","featured_image":"https:\/\/microbenotes.com\/wp-content\/uploads\/2020\/02\/Iron-Hematoxylin-Staining.jpg","author":{"name":"Faith Mokobi","avatar":"https:\/\/secure.gravatar.com\/avatar\/8b4f039c6d83a1b8509046a793299d54?s=96&d=mm&r=g"},"categories":[{"term_id":705,"name":"Staining","slug":"staining","term_group":0,"term_taxonomy_id":705,"taxonomy":"category","description":"<strong>Staining is a biochemical process usually conducted to increase the contrast of a microscopic image.<\/strong>\r\n<ul>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining samples with different dyes is an important part of microscopy where the color contrast of a magnified is increased to differentiate the stained portion of the image from the rest.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Microscopic techniques in hematology, histopathology, and clinical microbiology use staining as a method to study and diagnose diseases at the microscopic level.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Dyes used in staining can be used to define the biological tissues, cell populations, or subcellular organelles.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The purpose of staining might also differ as some staining procedures are used to differentiate between two groups of microorganisms (Gram Staining) while some are used to observe motility and presence of a capsule.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining is a part of biochemistry where a class-specific dye is added to a substrate to qualify or quantify the presence of a certain compound.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining, however, is not limited to biological samples, as it can be used to study the lamellar structure of polymers and the structure of block compounds.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Bright field microscopes are commonly used to observe stained cells or other biological materials to provide a distinction between the sample and the background.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">It is also difficult to observe colorless substances under a microscope, which is why the concept of staining is so popular.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining is of different types depending on their purpose; simple staining using a single stain to provide a contrast between the sample and the background; differential staining using more than one dyes in order to differentiate between cells or between organelles of a cell.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Among all the staining techniques, <a href=\"https:\/\/microbenotes.com\/gram-stain-principle-reagents-procedure-and-result-interpretation\/\" target=\"_blank\" rel=\"noopener\">Gram staining<\/a> is the most popular technique that are used to <a href=\"https:\/\/microbenotes.com\/differences-between-gram-positive-and-gram-negative-bacteria\/\" target=\"_blank\" rel=\"noopener\">differentiate between Gram-positive and Gram-negative microorganisms<\/a>.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Other staining techniques include <a href=\"https:\/\/microbenotes.com\/endospore-staining\/\" target=\"_blank\" rel=\"noopener\">endospore staining<\/a> for endospores, <a href=\"https:\/\/microbenotes.com\/acid-fast-stain-principle-reagents-procedure-and-result-interpretation\/\" target=\"_blank\" rel=\"noopener\">acid-fast staining<\/a> for fastidious organisms, <a href=\"https:\/\/microbenotes.com\/periodic-acid-schiff-pas-staining\/\" target=\"_blank\" rel=\"noopener\">Periodic-acid Schiff (PAS)<\/a> for biomolecules, and <a href=\"https:\/\/microbenotes.com\/romanowsky-stains\/\" target=\"_blank\" rel=\"noopener\">Romanowsky staining<\/a> to differentiate between <a href=\"https:\/\/microbenotes.com\/blood-cells\/\" target=\"_blank\" rel=\"noopener\">blood cells<\/a>.<\/span><\/li>\r\n<\/ul>","parent":0,"count":29,"filter":"raw","cat_ID":705,"category_count":29,"category_description":"<strong>Staining is a biochemical process usually conducted to increase the contrast of a microscopic image.<\/strong>\r\n<ul>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining samples with different dyes is an important part of microscopy where the color contrast of a magnified is increased to differentiate the stained portion of the image from the rest.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Microscopic techniques in hematology, histopathology, and clinical microbiology use staining as a method to study and diagnose diseases at the microscopic level.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Dyes used in staining can be used to define the biological tissues, cell populations, or subcellular organelles.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The purpose of staining might also differ as some staining procedures are used to differentiate between two groups of microorganisms (Gram Staining) while some are used to observe motility and presence of a capsule.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining is a part of biochemistry where a class-specific dye is added to a substrate to qualify or quantify the presence of a certain compound.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining, however, is not limited to biological samples, as it can be used to study the lamellar structure of polymers and the structure of block compounds.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Bright field microscopes are commonly used to observe stained cells or other biological materials to provide a distinction between the sample and the background.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">It is also difficult to observe colorless substances under a microscope, which is why the concept of staining is so popular.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Staining is of different types depending on their purpose; simple staining using a single stain to provide a contrast between the sample and the background; differential staining using more than one dyes in order to differentiate between cells or between organelles of a cell.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Among all the staining techniques, <a href=\"https:\/\/microbenotes.com\/gram-stain-principle-reagents-procedure-and-result-interpretation\/\" target=\"_blank\" rel=\"noopener\">Gram staining<\/a> is the most popular technique that are used to <a href=\"https:\/\/microbenotes.com\/differences-between-gram-positive-and-gram-negative-bacteria\/\" target=\"_blank\" rel=\"noopener\">differentiate between Gram-positive and Gram-negative microorganisms<\/a>.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Other staining techniques include <a href=\"https:\/\/microbenotes.com\/endospore-staining\/\" target=\"_blank\" rel=\"noopener\">endospore staining<\/a> for endospores, <a href=\"https:\/\/microbenotes.com\/acid-fast-stain-principle-reagents-procedure-and-result-interpretation\/\" target=\"_blank\" rel=\"noopener\">acid-fast staining<\/a> for fastidious organisms, <a href=\"https:\/\/microbenotes.com\/periodic-acid-schiff-pas-staining\/\" target=\"_blank\" rel=\"noopener\">Periodic-acid Schiff (PAS)<\/a> for biomolecules, and <a href=\"https:\/\/microbenotes.com\/romanowsky-stains\/\" target=\"_blank\" rel=\"noopener\">Romanowsky staining<\/a> to differentiate between <a href=\"https:\/\/microbenotes.com\/blood-cells\/\" target=\"_blank\" rel=\"noopener\">blood cells<\/a>.<\/span><\/li>\r\n<\/ul>","cat_name":"Staining","category_nicename":"staining","category_parent":0}]},"_links":{"self":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts\/24855"}],"collection":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/comments?post=24855"}],"version-history":[{"count":0,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts\/24855\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/media\/30282"}],"wp:attachment":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/media?parent=24855"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/categories?post=24855"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/tags?post=24855"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}