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Histological Techniques

Histological Techniques refer to the methods and procedures used to visualize, analyze, and study the microscopic structure and composition of biological tissues.
These techneiques involve the preparation, staining, and examination of tissue samples under a microscope, allowing researchers to observe cellular and subcellular features, identify tissue types, and detect pathological changes.
Histological techniques are essential tools in fields such as anatomy, pathology, and cell biology, contributing to our understanding of normal and diseased states.
They eanble the detailed examination of organ, tissue, and cellular morphology, facilitating the diagnosis, treatment, and research of a wide range of medical conditions.

Most cited protocols related to «Histological Techniques»

Histologic Assessment of α-Synuclein Pathology

Diagnostic histologic methods were performed on standard blocks of tissue that were fixed in 4% buffered formaldehyde and then either dehydrated and embedded in paraffin or cryoprotected and cut on a freezing, sliding microtome. Paraffin sections from the olfactory bulb and tract, anterior medulla (two levels anterior to the obex), anterior and mid-pons, mid-amygdala with adjacent transentorhinal area, anterior cingulate gyrus (1–3 cm posterior to the coronal slice containing the genu of the corpus callosum), middle temporal gyrus (at the level of the lateral geniculate nucleus), middle frontal gyrus (4–5 cm posterior to the frontal pole), and inferior parietal lobule were stained immunohistochemically for α-synuclein using a polyclonal antibody raised against an α-synuclein peptide fragment phosphorylated at serine 129, after epitope exposure with proteinase K. The process leading to the choice of immunohistochemical method, as well as details of the method, have been described in a previous publication (7 (link)). The density of α-synuclein-immunoreactive Lewy bodies and neurites in each of the above-mentioned brain regions was scored, for more than 90% of slides, by a single observer (TGB), without knowledge of diagnosis, as none, sparse, moderate, frequent and very frequent, using the templates provided by the Dementia with Lewy Bodies Consortium (66 (link)). The remaining slides were scored by trainees under the instruction of the primary observer. For the substantia nigra (SN), LTS was estimated using the same scoring method but on thioflavine-S-stained thick (40 micron) sections due to the standard laboratory practice of sectioning the SN in this manner for unbiased morphometric analysis.

Beach T.G., Adler C.H., Lue L., Sue L.I., Bachalakuri J., Henry-Watson J., Sasse J., Boyer S., Shirohi S., Brooks R., Eschbacher J., White CL I.I.I., Akiyama H., Caviness J., Shill H.A., Connor D.J., Sabbagh M.N, & Walker D.G. (2009). Unified Staging System for Lewy Body Disorders: Correlation with Nigrostriatal Degeneration, Cognitive Impairment and Motor Dysfunction. Acta neuropathologica, 117(6), 613-634.

Publication 2009

Amygdaloid Body Brain Corpus Callosum Dementia Diagnosis Endopeptidase K Epitopes Formaldehyde Gyrus, Anterior Cingulate Histological Techniques Immunoglobulins Knee Lateral Geniculate Body Lewy Bodies Medial Frontal Gyrus Medulla Oblongata Microtomy Middle Temporal Gyrus Neurites Olfactory Bulb Paraffin Paraffin Embedding Parietal Lobule Peptide Fragments Pons Serine SNCA protein, human Substantia Nigra thioflavine Tissues

Brain Tissue Donation for Lewy Body Disorders

Brain tissue was obtained from Sun Health Research Institute (SHRI), located in the Sun Cities retirement communities of northwest metropolitan Phoenix, Arizona. Brain necropsies were performed on elderly subjects who had volunteered for the SHRI Brain Donation Program (6 (link)). The majority of Brain Donation Program subjects are clinically characterized at SHRI with annual standardized test batteries that include functional, neuropsychological and neuromotor components, including the Mini Mental State Examination (MMSE) and Unified Parkinson’s Disease Rating Scale (UPDRS). Additionally, private medical records are requisitioned and reviewed for each subject and the postmortem Dementia Questionnaire (32 (link)) is administered to subject contacts to help determine the presence or absence of dementia for those subjects lacking standardized antemortem evaluations. The Brain Donation Program has been approved by the SHRI Institutional Review Board.
Subjects were chosen by searching the Brain Donation Program database for all cases with a completed neuropathologist’s examination, a full set of paraffin blocks (defined as blocks having all ten regions as defined in the histologic methods) and diagnoses of a Lewy body disorder, including PD, DLB, ILBD and ADLB. Comparison groups were composed of subjects without evidence of dementia or parkinsonism (normal elderly subjects) and subjects with AD but no Lewy body pathology (ADNLB).
Subjects received standardized neuropathological examinations. Specific diagnostic criteria were used for AD (1 (link)), PD (37 (link)) and DLB (67 (link)). For both AD and DLB, cases received the diagnosis if they were classified as “intermediate” or “high” probabilities in their respective classification schemes. Cases with LTS but not meeting these diagnostic criteria were designated as either ILBD, if they had no clinical history of parkinsonism or dementia, or ADLB if they had Alzheimer’s disease and Lewy bodies in any brain region but failed to meet criteria for DLB.
Gross and microscopic neuropathologic assessments were made by a single observer (TGB) without knowledge of the clinical history or clinical diagnosis; subsequently the clinical history was reviewed in order to make an appropriate clinicopathologic diagnosis. Subjects with Lewy body disorders were staged according to Braak (11 (link)) as well as the Dementia with Lewy Bodies Consortium (66 (link)). Apolipoprotein E genotyping was done on postmortem brain tissue using a standard technique (45 (link)).

Publication 2009

Aged Apolipoproteins E Autopsy Brain Dementia Diagnosis Ethics Committees, Research Histological Techniques Lewy Bodies Lewy Body Disease Microscopy Mini Mental State Examination Neuropathologist Paraffin Parkinsonian Disorders Physical Examination Tissues Wellness Programs

Dopamine Dynamics in Selectively-Bred Rats

The majority of these studies were conducted with adult male Sprague-Dawley rats from a selective-breeding colony which has been previously described14 (link). The data presented here were obtained from bHR and bLR rats from generations S18–S22. Equipment and procedures for Pavlovian conditioning have been described in detail elsewhere 13 (link),15 (link). Selectively-bred rats from generations S18, S20 and S21 were transported from the University of Michigan (Ann Arbor, MI) to the University of Washington (Seattle, WA) for the FSCV experiments. During each behavior session, chronically implanted microsensors, placed in the core of the nucleus accumbens, were connected to a head-mounted voltammetric amplifier for detection of dopamine by FSCV23 (link). Voltammetric scans were repeated every 100 ms to obtain a sampling rate of 10 Hz. Voltammetric analysis was carried out using software written in LabVIEW (National Instruments, Austin, TX). On completion of the FSCV experiments, recording sites were verified using standard histological procedures. To examine the effects of flupenthixol (Sigma, St. Louis, Missouri; dissolved in 0.9% NaCl) on the performance of sign-tracking and goal-tracking behavior, rats received an injection (i.p.) of 150, 300 or 600 μg/kg of the drug one hour prior to Pavlovian conditioning sessions 9, 11 and 13. Doses of the drug were counterbalanced between groups and interspersed with saline injections (i.p., 0.9% NaCl; prior to sessions 8, 10, 12 and 14) to prevent any cumulative drug effects. To examine the effects of flupenthixol on the acquisition of sign-tracking and goal-tracking behavior, rats received an injection of either saline (i.p.) or 225 μg/kg of the drug one hour prior to Pavlovian conditioning sessions 1–7.
Detailed methods and any associated references are available in the online version of the paper at www.nature.com/nature.

Flagel S.B., Clark J.J., Robinson T.E., Mayo L., Czuj A., Willuhn I., Akers C.A., Clinton S.M., Phillips P.E, & Akil H. (2010). A selective role for dopamine in reward learning. Nature, 469(7328), 53-57.

Publication 2010

Adult austin Dopamine Flupenthixol Head Histological Techniques Males Normal Saline Nucleus Accumbens Pharmaceutical Preparations Radionuclide Imaging Rats, Sprague-Dawley Rattus norvegicus Saline Solution

Melanocytic Lesions Test Cases

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Publication 2013

Biopsy Body Regions Eosin Histological Techniques Melanocyte Melanoma Patients Skin Youth

Whole-cell and cell-attached recordings in layer 6 auditory cortex

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Publication 2010

Adenosine Triphosphate, Magnesium Salt Agar Auditory Area Axon biocytin cesium chloride Cortex, Cerebral Egtazic Acid gluconate HEPES Histological Techniques Neurons Phocidae Phosphocreatine Plasma Membrane Psychological Inhibition Pyramidal Cells QX-314 Resting Potentials

Most recents protocols related to «Histological Techniques»

Histopathological Analysis of Renal Tissue

Renal tissue was fixed for at least 72 h in 5% buffered formaldehyde solution (Fischar), before dehydration in descending alcohol solution and embedding in paraffin (Thermo Fisher Scientific) were performed as described previously (18 (link)). For all histological staining procedures, 2 µm renal sections were prepared. Histopathological evaluation of renal and splenic tissue using periodic acid Schiff (PAS) staining were performed as described previously (18 (link)). Staining for kidney injury molecule-1 (KIM-1), BTK, lymphocyte antigen 6 complex (Ly6g), F4-80, CD3, Ki67 and cleaved caspase-3 (CC-3) were used to evaluate renal sections immunohistochemically. Generally, sections were deparaffinized and hydrated as described previously (18 (link)). Blocking of endogenous peroxidase was performed using 3% H₂O₂ (Carl Roth) and target retrieval solution (pH 6; Dako) was utilized for antigen retrieval in a pressure cooker. Bovine serum albumin (BSA; Sigma Aldrich) or 20% serum (PAA Laboratories) as well as avidin and biotin solution (15 min each; Vector Laboratories) were used each to block unspecific binding sites (Supplementary Table S3). Renal sections were incubated with primary antibody (Supplementary Table S4) overnight at 4°C. Sections were further incubated with secondary antibody (Supplementary Table S5) and with VectaStain ABC kit (Vector Laboratories) for 30 min each (Supplementary Table S3 for detailed information). As substrate, 3,3-diaminobenzidine (DAB; Vector Laboratories) was used and sections were counterstained with hemalaun (Carl Roth). Finally, renal sections were dehydrated and mounted for observation. Tris(hydroxymethyl)aminomethan (TRIS) buffer (pH 7.6) containing 50 mM TRIS (Carl Roth), 300 mM sodium chloride (Carl Roth), 0.04% Tween^® 20 (Sigma Aldrich) was used to wash renal sections between the staining processes. Staining of thrombocytes (glycoprotein-1b (GP1b)) and fibrin deposition (acid fuchsin–Orange G stain (SFOG)) was performed as described previously (18 (link), 19 (link)).

Kröller S., Wissuwa B., Dennhardt S., Krieg N., Thiemermann C., Daniel C., Amann K., Gunzer F, & Coldewey S.M. (2023). Bruton’s tyrosine kinase inhibition attenuates disease progression by reducing renal immune cell invasion in mice with hemolytic-uremic syndrome. Frontiers in Immunology, 14, 1105181.

Publication 2023

acid-fuchsin Antigens Avidin Binding Sites Biotin Blood Platelets Cardiac Arrest Caspase 3 Cloning Vectors Ethanol Fibrin Formalin Glycoproteins HAVCR1 protein, human Histological Techniques Immunoglobulins Kidney LD Antigens Orange G Periodic Acid Peroxidase Peroxide, Hydrogen Pressure Serum Serum Albumin, Bovine Sodium Chloride Spleen Stains Tissues Tromethamine Tween 20

Intestinal Morphometry Analysis in Birds

At the end of the study (12th week), 24 birds [six birds from each group (1 per replicate)] were selected for sample collection after 12 h fasting. The selected birds were euthanized with pentobarbital sodium (100 mg/kg BW) intravenously and cut open under aseptic conditions. The heart, liver, magnum and spleen of each bird were weighed and the organ index was calculated weight of organ (g)/body weight (g) × 100%. The jejunal samples of each bird were processed following that established in Gungor and Erener (2020) (link). About three-centimeter length of tissue from jejunum were flushed in physiological saline solution and placed in 10% buffered formalin, kept at 4°C for analysis.
The fixed jejunal samples were processed and stained with hematoxylin–eosin, while adopting the standard techniques for histology examination (Yamauchi et al., 2010 (link)). Images were observed with the aid of a microscope (An Olympus BX43 microscope; Olympus Corp., Tokyo, Japan). The villi morphometrics were obtained; ten intact villi of each sample and corresponding crypts were selected, measured, and the average value was obtained. The measurements were made with a software (Caseviewer Image). The measurements include; villi height (VH: measured from the top of the villus to the villus-crypt junction), the villus width (VW: was measured at the middle point of the villus), crypt depth (CD: from the base up to the crypt–villus transition region), the VH-CD ratio (measured as VH/CD), villi surface area (VSA: π × VW × VH) (Wang et al., 2016 (link); Thiam et al., 2021 (link)).

Obianwuna U.E., Qiu K., Wang J., Zhang H.J., Qi G.H., Huang L.L, & Wu S.G. (2023). Effects of dietary Clostridium butyricum and fructooligosaccharides, alone or in combination, on performance, egg quality, amino acid digestibility, jejunal morphology, immune function, and antioxidant capacity of laying hens. Frontiers in Microbiology, 14, 1125897.

Publication 2023

Asepsis Aves Body Weight CFC1 protein, human DNA Replication Eosin Formalin Heart Histological Techniques Jejunum Liver Microscopy Pentobarbital Sodium physiology Saline Solution Specimen Collection Spleen Tissues

Histopathological Assessment of Drosophila Neurodegeneration

Histopathological tests were performed using standard histological methods as previously described.^{78 (link),80 (link)-82 (link)} Briefly, on the day of histological assessments, Drosophila melanogaster flies were anesthetized with CO₂ (Fly CO₂ anesthesia setup; Genesee Scientific, 59-114/54-104M, USA), and placed on a CO₂ perfused pad for collecting. The flies were decapitated under a dissecting stereo microscope (Leica Microsystems, Leica M60 CMO), and the heads were fixed in 10% neutral buffered formalin (NBF) fixative at room temperature (22- 30°C) for 24 h, after which tissues were processed using routine histology techniques by dehydrating with graded alcohols, clearing with xylene, and embedding into paraffin wax using an automated tissue processor (Histokinette-SLEE MAINZ, MTP type). Every tissue was randomly sectioned into six, 6 μm-thick transverse histological sections using a rotary microtome (SLEE MAINZ, CUT4062), and the sections were then placed on slides and stained with hematoxylin and eosin (H &E) and combined Luxol fast blue (LFB) and Nissl (Klüver's) staining techniques following standard protocols.^{78 (link),80 (link)-82 (link)} The stained sections were mounted in mounting media and qualitative histological examination of the sections was done and photographed with a light microscope (Nikon Eclipse Ci-L Upright Microscope, New York, USA), at a magnification of 200x or 400x, digital camera (Nikon DS-Fi1c Digital Camera, New York, USA), and imaging software (Nikon NIS- NIS-Elements F Ver4.60.00 Imaging software), for image analysis and documentation.
A qualitative examination of the tissues was done using previously described methods^{64 ,83 (link)-85 (link)} where brain neurodegeneration was categorized as normal, moderate, or severe based on the size and frequency of brain vacuolations in H and E stained-sections following previous methods.^{84 (link),85 (link)} The non-myelinating Schwann cells in mammals are comparable to the ‘ensheathing’ glial cells within the CNS of Drosophila which encase axons and neuropil of the flies,^{86 (link),87 (link)} therefore, LFB in Klüver LFB stain was used to demonstrate axonal degeneration of neurons rather than axonal demyelination.^{88 (link)} The nature of the Nissl substance and nerve tracts (axons) were demonstrated using the Klüver LFB-stained tissues following previous methods,^{64 ,83 (link)} where the nerve tracts were shown by blue color and the Nissl substance was shown by magenta (violet) colour. A weak LFB stain (light blue patchy areas) indicated axonal degeneration of nerve tracts, while a weak cresyl echt violet stain (light violet) indicated abnormalities in Nissl substance,^{64 ,83 (link)}

supplementary file 2

Ssempijja F., Dare S.S., Bukenya E.E., Kasozi K.I., Kenganzi R., Fernandez E.M, & Vicente-Crespo M. (2023). Attenuation of Seizures, Cognitive Deficits, and Brain Histopathology by Phytochemicals of Imperata cylindrica (L.) P. Beauv (Poaceae) in Acute and Chronic Mutant Drosophila melanogaster Epilepsy Models. Journal of Evidence-based Integrative Medicine, 28, 2515690X231160191.

Publication 2023

Alcohols Anesthesia Axon Brain Congenital Abnormality cresyl violet Debility Demyelination Diptera Drosophila Drosophila melanogaster Eosin Fingers Fixatives Formalin Head Histological Techniques Light Light Microscopy Luxol Fast Blue MBS Mammals Microscopy Microtomy Nerve Degeneration Nervousness Neuroglia Neuropil Rosaniline Dyes Schwann Cells Staining Stains Tissues Tissue Stains Viola Xylene

Histological Analysis of Skin Wound Healing

After the experiments, the experimented mouse or pig skin was cut off. The targeted area was the incised skin. Skin tissues were embedded in the Tissue‐Tek optimal cutting temperature compound and cryosectioned into 5‐µm slices. The tissue slabs were processed by standard histological procedures, histochemically stained with hematoxylin and eosin (H&E), F4/80 antibody, trichrome, and CD³⁺ antibody.^[¹⁰⁵, ¹⁰⁶^] Antibodies were diluted according to manufacturer's instruction, unless indicated otherwise. Serving as a control, two wounds were created on the dorsal side of each mouse by removing full‐thickness skin via 3‐mm punch biopsy. Tegaderm films (3 M Inc.) were used to cover the wounds and prevent water loss in all the mice until the wounds were fully epithelialized. At days 7 and 14 postsurgery, three mice in each group were euthanized and the wounded skin removed, fixed in formalin, embedded in paraffin, and sectioned. Trichrome, F4/80 antibody, and CD³⁺ antibody staining were used for histological observations. Microscopic evaluation of the tissue sections was performed after that. The sections with F4/80, and CD³⁺ staining were observed under the microscope (Eclipse Ti2; Nikon Inc.) at 20× and 200× magnification. Four fields were randomly selected from each section to count the F4/80 positive macrophages, and CD³⁺ positive T‐cells. Immunoreactive cells were quantified as the mean cell count expressing the appropriate positive marker per high‐power field (HPF). Histological data were expressed as mean ± standard deviation (SD). Statistical analysis was performed by Student's t‐test. A p value of < 0.05 was considered significant.

Zhang J., Zheng Y., Lee J., Hoover A., King S.A., Chen L., Zhao J., Lin Q., Yu C., Zhu L, & Wu X. (2023). Continuous Glucose Monitoring Enabled by Fluorescent Nanodiamond Boronic Hydrogel. Advanced Science, 10(7), 2203943.

Publication 2023

Antibodies Biopsy Cells Eosin Formalin Histological Techniques Immunoglobulins Macrophage Microscopy Mus Paraffin Embedding Skin T-Lymphocyte Tissues

Visualizing Radiation Dermatitis in Multicellular Model

Since in this pre-clinical study we aim to visualise radiation-induced lesions, alterations, and suspicious features in the multicellular model that are predictive of the development of radiation dermatitis, we used both non-invasive OCT imaging and a comparative histological method, although the latter requires biopsy/sectioning and staining.

Bromberger L., Heise B., Felbermayer K., Leiss-Holzinger E., Ilicic K., Schmid T.E., Bergmayr A., Etzelstorfer T, & Geinitz H. (2023). Radiation-induced alterations in multi-layered, in-vitro skin models detected by optical coherence tomography and histological methods. PLOS ONE, 18(3), e0281662.

Publication 2023

Biopsy Histological Techniques Radiation Radiation-Induced Dermatitis

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More about "Histological Techniques"

Histological techniques refer to the methodologies and procedures employed to visualize, analyze, and study the microscopic structure and composition of biological tissues.
These techniques involve the preparation, staining, and examination of tissue samples under a microscope, enabling researchers to observe cellular and subcellular features, identify tissue types, and detect pathological changes.
Histological techniques are essential tools in fields such as anatomy, pathology, and cell biology, contributing to our understanding of normal and diseased states.
They enable the detailed examination of organ, tissue, and cellular morphology, facilitating the diagnosis, treatment, and research of a wide range of medical conditions.
The histological process often begins with tissue fixation, using chemicals like paraformaldehyde or formalin to preserve the cellular structure.
The fixed tissue is then dehydrated, embedded in paraffin wax, and sectioned using a microtome to create thin slices.
These sections are then stained with dyes like hematoxylin and eosin, which selectively bind to different cellular components, enhancing contrast and allowing for the identification of various tissue types and structures.
Light microscopy techniques, such as those utilized in the Image-Pro Plus 6.0, BX51 microscope, and Eclipse 80i, are widely employed in histological analysis.
These advanced microscopes enable the detailed examination of tissue samples, revealing intricate cellular features and patterns.
The BX41 microscope, for instance, is commonly used in histopathology for the diagnosis of diseases by analyzing tissue samples.
Histological techniques also involve the use of fluorescent stains, such as DAPI, which bind to DNA and allow for the visualization of nuclei and chromosomes.
These techniques are particularly useful in cell biology research, as they enable the study of cellular processes and structures at a subcellular level.
Ultimately, histological techniques are indispensable tools in the fields of medicine, biology, and research, providing invaluable insights into the structure and function of living organisms and contributing to our understanding of health and disease.