Application of anti-BCG antibody for rapid immunohistochemical detection of bacteria, fungi and protozoa in formalin-fixed paraffin-embedded tissue samples

2008 ◽  
Vol 56 (1) ◽  
pp. 89-99 ◽  
Author(s):  
Levente Szeredi ◽  
Róbert Glávits ◽  
Miklós Tenk ◽  
Szilárd Jánosi
Keyword(s):  
Bacterial Species ◽  
Histochemical Staining ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Bacteriological Culture ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed Paraffin Embedded ◽  
Almost All ◽  
Formalin Fixed

The applicability of an anti- Mycobacterium bovis (BCG) antibody-based immunohistochemistry (IHC) procedure was investigated using everyday veterinary pathological samples collected from 13 different animal species. Fifty-one formalin-fixed and paraffin-embedded tissue samples were selected for this study. Forty, 4 and 7 tissue samples contained different species of bacteria, fungi and protozoa, respectively. Three serial sections were prepared in each case. Two sections were pre-treated with enzyme and heat, respectively, while the last section was not pre-treated. In seven cases the sensitivity of histochemical staining (HSM), IHC and bacteriological culture were compared. Heating of the sections in a microwave oven was the most effective method in the case of almost all pathogens used. Strong or moderate positive reactions were observed for 26 bacterial species, all fungal and 2 protozoal species, while weak reactions occurred for 2 bacterial and 1 protozoal species. Only 4 protozoal and 12 bacterial species, including Leptospira and all the five Mycoplasma species examined, showed no reaction in this test. IHC had almost the same sensitivity as bacteriological culture and was more sensitive than HSM. The IHC method presented here should be preferred to HSM as a general screening tool in cases where pathological lesions suspicious for infections are evident and no microorganism can be cultured in vitro or only formalin-fixed tissue samples are available for the laboratory examination.

Effect of formalin fixation on thermal conductivity of the biological tissue

2014 ◽  
Author(s):  
Steven C. Bauserman ◽  
Jonathan W. Valvano
Keyword(s):  
Thermal Conductivity ◽  
Biological Tissues ◽  
Formalin Fixation ◽  
Fatty Tissue ◽  
Fresh Tissue ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed

Effect of formalin fixation on thermal conductivity of the biological tissues is presented. A self-heated thermistor probe was used to measure the tissue thermal conductivity. The thermal conductivity of muscle and fatty tissue samples was measured before the formalin fixation and then 27 hours after formalin fixation. The results indicate that the formalin fixation does not cause a significant change in the tissue thermal conductivity of muscle and fatty tissues. In the clinical setting, tissues removed surgically are often fixed in formalin for subsequent pathological analysis. These results suggest that, in terms of thermal properties, it is equally appropriate to perform in vitro studies in either fresh tissue or formalin-fixed tissue.

Abstract 4281: Targeted or whole genome sequencing of formalin-fixed tissue samples

2014 ◽  
Author(s):  
Sarah Munchel ◽  
Yen Hoang ◽  
Yue Zhao ◽  
Joseph Cottrell ◽  
Brandy Klotzle ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Whole Genome ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed

scholarly journals Specific Detection of Lawsonia intracellularis in Porcine Proliferative Enteropathy Inferred from Fluorescent rRNA In Situ Hybridization

1998 ◽  
Vol 35 (2) ◽  
pp. 153-156 ◽  
Author(s):  
M. Boye ◽  
T. K. Jensen ◽  
K. Møller ◽  
T. D. Leser ◽  
S. E. Jorsal
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Lawsonia Intracellularis ◽  
Specific Detection ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Formalin Fixed Tissue ◽  
Proliferative Enteropathy ◽  
Formalin Fixed

Fluorescent in situ hybridization targeting 16S ribosomal RNA was used for specific detection of the obligate intracellular bacterium Lawsonia intracellularis in enterocytes from pigs affected by proliferative enteropathy. A specific oligonucleotide probe was designed and the specificity of the probe was determined by simultaneous comparison with indirect immunofluorescence assay for detection of L. intracellularis in formalin-fixed tissue samples from 15 pigs affected by porcine proliferative enteropathy. We used 10 tissue samples from pigs without proliferative mucosal changes as negative controls. The results showed that the oligonucleotide probe is specific for L. intracellularis and that fluorescent in situ hybridization targeting ribosomal RNA is a suitable and fast method for specific detection and histological recognition of L. intracellularis in formalin-fixed tissue.

A Basis for Distinguishing Cultured Dendritic Cells and Macrophages in Cytospins and Fixed Sections

2005 ◽  
Vol 8 (1) ◽  
pp. 43-51 ◽  
Author(s):  
Jukka Vakkila ◽  
Michael T. Lotze ◽  
Connie Riga ◽  
Ronald Jaffe
Keyword(s):  
Dendritic Cells ◽  
Cell Types ◽  
Fixed Tissue ◽  
Reliable Identification ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Hla Dr ◽  
Formalin Fixed

There is a burgeoning literature on the contrasting role of intratumoral dendritic cells (DCs) and tumor-associated macrophages, making reliable identification of both cell types in clinical and experimental tissue sections important. However, because these cell types are closely related and share several differentiation antigens, their absolute distinction in tissue sections is difficult. We differentiated DCs and macrophages from monocytes in vitro, prepared cytospins and paraffin-embedded sections of the various cell populations, and tested a variety of antibodies that purportedly recognize monocytes and DCs for their capacity to react and distinguish cells after conventional formalin fixation. Cultured DCs but not macrophages were detected by fascin, DC-LAMP, and CD83 with a predictable increase in staining that paralleled their maturation. Staining by CD1a was found on immature DCs but was weak and absent on mature DCs and macrophages, respectively. CD14 and CD163 were characteristic for macrophages and absent on DCs. CD68, HLA-DR, and S100 did not discriminate between DCs and macrophages. We conclude that antigens such as HLA-DR and S100 are not in themselves sufficient for identification of DCs in formalin-fixed tissue sections, but that additional macrophage-specific (CD14, CD163) and DC-specific (CD1a, CD83, fascin, DC-LAMP) antigens should be used to distinguish cell types from each other and to provide information on their state of maturation.

scholarly journals Biochemical Subtyping of Amyloid in Formalin-Fixed Tissue Samples Confirms and Supplements Immunohistologic Data

2004 ◽  
Vol 121 (6) ◽  
pp. 794-800 ◽  
Author(s):  
Batia Kaplan ◽  
Brian M. Martin ◽  
Avi Livneh ◽  
Mordechai Pras ◽  
Gloria R. Gallo
Keyword(s):  
Fixed Tissue ◽  
Tissue Samples ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed

scholarly journals Resin Tissue Microarrays: a Universal Format for Immunohistochemistry

2005 ◽  
Vol 53 (10) ◽  
pp. 1189-1197 ◽  
Author(s):  
William J. Howat ◽  
Anthony Warford ◽  
Joanne N. Mitchell ◽  
Kay F. Clarke ◽  
Jen S. Conquer ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Tissue Microarrays ◽  
Single Chain ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Glycol Methacrylate ◽  
Formalin Fixed Tissue ◽  
Single Chain Fv ◽  
Nuclear Antigens ◽  
Formalin Fixed

Tissue microarray (TMA) technology allows the miniaturization and characterization of multiple tissue samples on a single slide and commonly uses formalin-fixed paraffin-embedded (FFPE) tissue or acetone-fixed frozen tissue. The former provides good morphology but can compromise antigenicity, whereas the latter provides compromised morphology with good antigenicity. Here, we report the development of TMAs in glycol methacrylate resin, which combine the advantages of both methods in one embedding format. Freshly collected tissue fixed in -20C acetone or 10% neutral buffered formaldehyde were cored and arrayed into an intermediary medium of 2% agarose before infiltration of the agarose array with glycol methacrylate resin. Acetone-fixed resin TMA demonstrated improved morphology over acetone-fixed frozen TMA, with no loss of antigenicity. Staining for extracellular, cell surface, and nuclear antigens could be realized with monoclonal and polyclonal antibodies as well as with monomeric single-chain Fv preparations. In addition, when compared with FFPE TMA, formalin-fixed tissue in a resin TMA gave enhanced morphology and subcellular detail. Therefore, resin provides a universal format for the construction of TMAs, providing improved tissue morphology while retaining antigenicity, allows thin-section preparation, and could be used to replace preparation of frozen and FFPE TMAs for freshly collected tissue.

scholarly journals Constant Detection of CD2, CD3, CD4, and CD5 in Fixed and Paraffin-embedded Tissue Using the Peroxidase-mediated Deposition of Biotin-Tyramide

1997 ◽  
Vol 45 (12) ◽  
pp. 1665-1672 ◽  
Author(s):  
Rainer Malisius ◽  
Hartmut Merz ◽  
Boris Heinz ◽  
Evariste Gafumbegete ◽  
Britta U. Koch ◽  
...  
Keyword(s):  
Binding Sites ◽  
Lymphoid Tissues ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Leukocyte Antigens ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Immunohistochemical Techniques ◽  
Microwave Techniques ◽  
Formalin Fixed

Immunohistochemical methods are widely used for diagnostic purposes in histopathology. However, the use of most monoclonal anti-leukocyte antibodies is limited to frozen tissues. Initially, it was believed that formalin fixation in particular, which is the gold standard for morphological tissue preservation, destroys most of the antigen binding sites. In recent years, protease digestion and the introduction of microwave techniques have significantly enhanced the sensitivity of immunohistochemical techniques, and a variety of hidden antigen sites in formalin-fixed tissue have been retrieved for initially unreactive antibodies. It therefore became clear that many of the leukocyte antigens are not irreversibly destroyed but are most probably masked during the fixation process. We developed a technique combining optimized pretreatment of formalin-fixed tissue with a dramatic enhancement of the immunohistochemical sensitivity and named it the ImmunoMax method. The ImmunoMax method proves that by optimizing the technique at the following three levels it is possible to detect formalin-sensitive leukocyte antigens: (a) standard fixation of the tissue; (b) sufficient antigen unmasking; and (c) increasing the substrate turnover by multiplication of binding sites with subsequent enhancement of the immunohistochemical reaction. Using this optimized ImmunoMax method, we were able to detect CD2, CD3, CD4, and CD5 with conventional monoclonal antibodies in formalin-fixed, paraffin-embedded tissue specimens of various lymphoid tissues.

scholarly journals Effects of fixation time and enzymatic digestion on immunohistochemical demonstration of bromodeoxyuridine in formalin-fixed, paraffin-embedded tissue.

1988 ◽  
Vol 36 (5) ◽  
pp. 511-514 ◽  
Author(s):  
Y Hayashi ◽  
M Koike ◽  
M Matsutani ◽  
T Hoshino
Keyword(s):  
Human Brain ◽  
Enzymatic Digestion ◽  
Fixation Time ◽  
Reaction Products ◽  
Human Brain Tumor ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

We studied the effects of fixation time and enzymatic digestion on immunohistochemical staining for bromodeoxyuridine (BUdR) in excised rat and human gastrointestinal tissues and human brain tumors which had been fixed in formalin after intravenous administration of BUdR shortly before biopsy of tissue. In formalin-fixed rat gastrointestinal tissues not treated with proteinase, the reaction products were insufficient to identify BUdR-positive cells. Results similar to those in ethanol-fixed tissue were obtained when formalin-fixed tissue sections were treated with protease, pepsin, or trypsin. The longer the material had been fixed in formalin, the longer the incubation in proteinase required to identify BUdR-labeled nuclei. The BUdR labeling indices of formalin-fixed human brain tumor specimens treated with protease were comparable to those of ethanol-fixed tissues. Sufficient BUdR staining was obtained even in tissues fixed in formalin for prolonged periods. Therefore, the BUdR labeling index can be determined retrospectively in clinical materials stored in formalin.

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