Tissue preparation, immunohistochemistry, Imaging, and Quantification v1

Author(s):  
Bryan Yoo ◽  
Jessica Griffiths ◽  
Sarkis Mazmanian

Protocol for neuronal imaging used in Yoo et al 2021

Author(s):  
Kenjiro Yasuda

Localization of amylase,chymotrypsinogen and trypsinogen in pancreas was demonstrated by Yasuda and Coons (1966), by using fluorescent antibody method. These enzymes were naturally found in the zymogen granules. Among them, amylase showed a diffuse localization around the nucleus, in addition to the zymogen granules. Using ferritin antibody method, scattered ferritin granules were also found around the Golgi area (Yasuda et al.,1967). The recent advance in the tissue preparation enables the antigen to be localized in the ultrathin frozen sections, by applying the labeled antibodies onto the sections instead of staining the tissue en bloc.The present study deals with the comparison of the localization of amylase and lipase demonstrated by applying the bismuth-labeled, peroxidase-labeled and ferritin-labeled antibody methods on the ultrathin frozen sections of pancreas, and on the blocks of the same tissue.


Author(s):  
Anne F. Bushnell ◽  
Sarah Webster ◽  
Lynn S. Perlmutter

Apoptosis, or programmed cell death, is an important mechanism in development and in diverse disease states. The morphological characteristics of apoptosis were first identified using the electron microscope. Since then, DNA laddering on agarose gels was found to correlate well with apoptotic cell death in cultured cells of dissimilar origins. Recently numerous DNA nick end labeling methods have been developed in an attempt to visualize, at the light microscopic level, the apoptotic cells responsible for DNA laddering.The present studies were designed to compare various tissue processing techniques and staining methods to assess the occurrence of apoptosis in post mortem tissue from Alzheimer's diseased (AD) and control human brains by DNA nick end labeling methods. Three tissue preparation methods and two commercial DNA nick end labeling kits were evaluated: the Apoptag kit from Oncor and the Biotin-21 dUTP 3' end labeling kit from Clontech. The detection methods of the two kits differed in that the Oncor kit used digoxigenin dUTP and anti-digoxigenin-peroxidase and the Clontech used biotinylated dUTP and avidinperoxidase. Both used 3-3' diaminobenzidine (DAB) for final color development.


Author(s):  
J.D. Shelburne ◽  
G.M. Roomans

Proper preparative procedures are a prerequisite for the validity of the results of x-ray microanalysis of biological tissue. Clinical applications of x-ray microanalysis are often concerned with diagnostic problems and the results may have profound practical significance for the patient. From this point of view it is especially important that specimen preparation for clinical applications is carried out correctly.Some clinical problems require very little tissue preparation. Hair, nails, and kidney and gallbladder stones may be examined and analyzed after carbon coating. High levels of zinc or copper in hair may be indicative of dermatological or systemic diseases. Nail clippings may be analyzed (as an alternative to the more conventional sweat test) to confirm a diagnosis of cystic fibrosis. X-ray microanalysis in combination with scanning electron microscopy has been shown to be the most reliable method for the identification of the components of kidney or gallbladder stones.A quantitatively very important clinical application of x-ray microanalysis is the identification and quantification of asbestos and other exogenous particles in lung.


Author(s):  
K. Florian Klemp ◽  
J.R. Guyton

The earliest distinctive lesions in human atherosclerosis are fatty streaks (FS), characterized initially by lipid-laden foam cell formation. Fibrous plaques (FP), the clinically significant lesions, differ from FS in several respects. In addition to foam cells, the FP also exhibit fibromuscular proliferation and a necrotic core region rich in extracellular lipid. The possible transition of FS into mature FP has long been debated, however. A subset of FS described by Katz etal., was intermediate in lipid composition between ordinary FS and FP. We investigated this hypothesis by electron microscopic cytochemistry by employing a tissue processing technique previously described by our laboratory. Osmium-tannic acid-paraphenylenediamine (OTAP) tissue preparation enabled ultrastructural analysis of lipid deposits to discern features characteristic of mature fibrous plaques.


Author(s):  
K. N. Colonna ◽  
G. Oliphant

Harmonious use of Z-contrast imaging and digital image processing as an analytical imaging tool was developed and demonstrated in studying the elemental constitution of human and maturing rabbit spermatozoa. Due to its analog origin (Fig. 1), the Z-contrast image offers information unique to the science of biological imaging. Despite the information and distinct advantages it offers, the potential of Z-contrast imaging is extremely limited without the application of techniques of digital image processing. For the first time in biological imaging, this study demonstrates the tremendous potential involved in the complementary use of Z-contrast imaging and digital image processing.Imaging in the Z-contrast mode is powerful for three distinct reasons, the first of which involves tissue preparation. It affords biologists the opportunity to visualize biological tissue without the use of heavy metal fixatives and stains. For years biologists have used heavy metal components to compensate for the limited electron scattering properties of biological tissue.


2017 ◽  
pp. 437-446
Author(s):  
Maria Ciesielska

Men’s circumcision is in many countries considered as a hygienic-cosmetic or aesthetic treatment. However, it still remains in close connection with religious rites (Judaism, Islam) and is still practiced all over the world. During the Second World War the visible effects of circumcision became an indisputable evidence of being a Jew and were often used especially by the so-called szmalcownicy (blackmailers). Fear of the possibility of discovering as non-Aryan prompted many Jews hiding on the so-called Aryan side of Warsaw to seek medical practitioners who would restore the condition as it was before the circumcision. The reconstruction surgery was called in surgical jargon “knife baptizing”. Almost all of the procedures were performed by Aryan doctors although four cases of hiding Jewish doctors participating in such procedures are known. Surgical technique consisted of the surgical formation of a new foreskin after tissue preparation and stretching it by manual treatment. The success of the repair operation depended on the patient’s cooperation with the doctor, the worst result was in children. The physicians described in the article and the operating technique are probably only a fragment of a broader activity, described meticulously by only one of the doctors – Dr. Janusz Skórski. This work is an attempt to describe the phenomenon based on the very scanty source material, but it seems to be the first such attempt for several decades.


2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.


2008 ◽  
Vol 2008 (12) ◽  
pp. pdb.prot5088-pdb.prot5088 ◽  
Author(s):  
K. Pang ◽  
M. Q. Martindale
Keyword(s):  

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