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Tissue Fixation

Tissue Fixation is the process of preserving biological tissues to maintain their structural integrity and chemical composition.
This is a crucial step in many research and diagnostic applications, such as histology, microscopy, and immunohistochemistry.
The fixation process typically involves the use of chemical agents, such as formalin or glutaraldehyde, which cross-link proteins and stabilize cellular components.
Proper tissue fixation is essential for accurate analysis and reproducible results.
Researchers can leverage AI-powered platforms like PubCompare.ai to easily locate, compare, and identify the best tissue fixation protocols from the literature, preprints, and patents, enhancing research efficiecy and confidence.

Most cited protocols related to «Tissue Fixation»

1
Constitutive and Conditional Islet1 Knockout Mouse Generation
Cited 19 times
Mice bearing a constitutive null allele of Islet1 were a gift of Sam Pfaff 7 (link). The Brn3atauLacZ mouse line has been previously described18 (link). Generation of Islet1MCM (MerCreMer) mice has also been reported25 (link). Details of the generation of Islet1F mice will be reported elsewhere. Briefly, a genomic fragment of encompassing exon 4 of mouse Isl1 gene was cloned and a Neo-selectable targeting construct was generated in which this fragment is flanked by loxP sites (Fig. S1). Embryonic stem (ES) cells were electroporated with this construct and neomycin-resistant ES cell clones were screened for correct targeting of the Isl1 locus by Southern analysis. Two recombinant clones were used for the blastocyst injection and chimeric mice were crossed to C57BL/6J females to generate heterozygous mice (IsletF/+). The Neomycin resistance gene was removed by crossing IsletF/+ mice to a FLPeR deleter strain (Supplementary Methods online). Islet1F/+ mice were intercrossed to generate homozygous floxed Islet1 mice (Islet1F/F).
Methods for genetic crosses, tamoxifen-induced Islet1 excision, tissue fixation, Xgal staining, immunostaining, in situ hybridization, microarray and Q-PCR analysis and in situ hybridization appear in the Supplementary Methods online. Primers for conventional and real-time genotyping of the floxed Isl1 and Wnt1-cre alleles appear in Table S3.
Sun Y., Dykes I.M., Liang X., Eng S.R., Evans S.M, & Turner E.E. (2008). A central role for Islet1 in sensory neuron development linking sensory and spinal gene regulatory programs. Nature neuroscience, 11(11), 1283-1293.
Publication 2008
Alleles Blastocyst Chimera Clone Cells Embryonic Stem Cells Exons Females Genes Genes, vif Genome Heterozygote Homozygote In Situ Hybridization Microarray Analysis Mus Neomycin Oligonucleotide Primers Strains Tamoxifen Tissue Fixation WNT1 protein, human
2
Optimizing Prostate Cancer Treatment Planning
Cited 15 times
MRI and CT scans for treatment planning were performed 7 days after fiducial implantation to ensure adequate tissue fixation and to allow for resolution of procedural edema/inflammation (Poggi et al., 2003 (link); Pouliot et al., 2003 (link); Kupelian et al., 2005 (link)). Precautions were taken to minimize prostate motion during the planning scans and treatment. Specifically, starting 5 days prior to acquisition of the planning scans until the end of treatment, patients maintained a low fiber diet to reduce intestinal gas (Smitsmans et al., 2008 (link)). Fasts were also initiated 4 h before both acquisition of the planning scans and each treatment session to minimize rectal movement. Enemas were performed 2 h prior to acquisition of the planning scans and each treatment session to minimize rectal volume. All patients were imaged and treated in the supine treatment position with a knee cushion to maximize patient comfort and limit prostate motion in response to respiration (Malone et al., 2000 (link)).
Fused thin cut CT images (1.25 mm) and high-resolution MR images were used for treatment planning (Figures 3A–C). MRI imaging was employed to define the target volume (Roach et al., 1996 (link)) and to aid in accurate localization of the bladder neck, membranous urethra, and penile bulb (Mclaughlin et al., 2005 (link)). Intra-prostatic fiducials were employed to guide image co-registration and limit fusion errors (Parker et al., 2003 (link)). MR images were obtained on a 1.5-T scanner in a phased-array torso coil. The MRI was quickly (<1 h) followed by a CT scan to minimize anatomical changes in the rectum that may interfere with image fusion. Two MRI sequences were employed to maximize visualization of the fiducials (susceptibility-weighted gradient-echo images) and the soft tissues (axial high-resolution turbo T2-weighted spin-echo images). Treatment planning included the prostate as the gross target volume (GTV). The CTV included the prostate and the proximal seminal vesicles. The PTV equaled the CTV expanded by 3 mm posteriorly and 5 mm in all other dimensions. Treatment planning was generally completed within 1 week of imaging and subsequent treatment was initiated within a 1- to 2-week window.
Lei S., Piel N., Oermann E.K., Chen V., Ju A.W., Dahal K.N., Hanscom H.N., Kim J.S., Yu X., Zhang G., Collins B.T., Jha R., Dritschilo A., Suy S, & Collins S.P. (2011). Six-Dimensional Correction of Intra-Fractional Prostate Motion with CyberKnife Stereotactic Body Radiation Therapy. Frontiers in Oncology, 1, 48.
Publication 2011
ECHO protocol Edema Enema Fibrosis Inflammation Intestines Knee Medulla Oblongata Movement Neck Ovum Implantation Patients Penis Prostate Radionuclide Imaging Rectum Respiration Seminal Vesicles Susceptibility, Disease Therapy, Diet Tissue, Membrane Tissue Fixation Tissues Torso Urethra Urinary Bladder X-Ray Computed Tomography
3
Tissue Fixation and Immunohistochemistry of Mouse Eyes
Cited 12 times
Tissue fixation and sectioning were performed as previously described [34] (link). Briefly, eyes from mice at 2 days post-injection were enucleated and fixed in 4% paraformaldehyde/PBS for 16 h 4°C prior to paraffin embedding. Tissue sections (10 µm thickness) were deparaffinized, rehydrated, and blocked in 5% BSA/PBS for 30 min at room temperature (RT). Slides were briefly washed with PBS and incubated with anti-GFP (Molecular Probes Inc.) at 1∶1000 in 1× BSA/PBS for 2 h at RT. Following a brief washing in PBS, vectashield with DAPI (Vector Laboratories Inc.) was applied and the slide was coverslipped. Sections were viewed at room temperature with an Axioskop 50 (Carl Zeiss Inc.) in the autoexpose mode using a ×20, ×40, or ×63 objective. Images were captured with an Axiocam HR digital camera (Carl Zeiss Inc.) using Axiovision 3.1 software (Carl Zeiss Inc.).
Farjo R., Skaggs J., Quiambao A.B., Cooper M.J, & Naash M.I. (2006). Efficient Non-Viral Ocular Gene Transfer with Compacted DNA Nanoparticles. PLoS ONE, 1(1), e38.
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Publication 2006
Cloning Vectors DAPI Eye Fingers Molecular Probes Mus paraform Tissue Fixation Tissues
4
Tissue Preparation for RNA and Histology
Cited 10 times
Organs were prepared from adult Wistar rats, cut to appr. 3–5 mm thick pieces, and immediately submerged in 10% neutral buffered formalin for fixation and embedding in paraffin, or stabilized in RNAlater solution (QIAGEN GmbH, Hilden, Germany), for reference samples with ideal RNA preservation, respectively. Formalin fixation of tissue specimens was performed over night (20–24 h) at room temperature, unless otherwise noted. Specimens were dehydrated in 70%, 90%, and 100% ethanol, followed by xylene, for 2×1 h each solution, then embedded in Paraplast Plus (Sherwood Medical Co., St. Louis) for 2–3 h at 60°C. For RNA isolation, 10 µm sections were cut on a standard microtome. In each case, the first 2 sections were discarded to exclude negative effects from exposure to air.
FFPE specimens from surgically removed lung carcinoma were obtained using routine diagnostic procedures; the study was approved by the ethical committee of the Ärztekammer Hamburg.
von Ahlfen S., Missel A., Bendrat K, & Schlumpberger M. (2007). Determinants of RNA Quality from FFPE Samples. PLoS ONE, 2(12), e1261.
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Publication 2007
Adult Biologic Preservation Diagnostic Tests, Routine Ethanol Formalin isolation Lung Cancer Microtomy Operative Surgical Procedures Rats, Wistar Tissue Fixation Xylene
5
Immunofluorescence Staining and Microscopy Protocol
Cited 9 times
For fixation of cells 4% paraformaldehyde in phosphate buffered saline (PBS, pH 7.4), precooled to 4°C, was added to the samples and incubated for 20 min. at room temperature. Permeablization was with 0.2% Triton X-100 in PBS for 20 min. at room temperature. For fixation of tissues 4% paraformaldehyde and 0.5% Triton X-100 in PBS, prewarmed to 37°C, was added to the samples and incubated for 20 min. at room temperature. No further permeabilization was performed. For both sample types non-specific binding was blocked by incubation in 4% bovine serum albumin (BSA; fraction V, Fisher Scientific), 2% donkey serum, 0.1% Triton X-100 in PBS for 1 hr. at room temperature. Antibodies were diluted into the above blocking solution at saturating concentrations (Table S1) and incubated for 2 hr. at room temperature (cells) or overnight at 4°C (tissue) with rocking in a humidified chamber. Secondary antibodies, when used, were also diluted in blocking solution and incubated for 2 hr. at room temperature. Nuclei were counterstained with 2 μg/ml DAPI in PBS, 0.2% Triton X-100 for 15 min. (cells) or 45 min. (tissues).
Cell culture images were acquired using a Zeiss Axiovert 200M fluorescence microscope equipped with a Roper CoolSnap HQ monochrome camera controlled by MetaMorph software (Molecular Devices). Tissue sections were imaged on a Zeiss LSM 710 Confocal Laser Scanning Microscope. A z-series encompassing the full thickness of the tissue was collected for each field. All microscope settings and exposure times were set to collect images below saturation and were kept constant for all images taken in one experiment. Image analysis was performed using either MetaMorph software or ImageJ open source software from the NIH (http://rsbweb.nih.gov/ij/). The region of each nucleus in an image was defined using the DAPI channel (435–485 nm emission), and the total and average fluorescence intensities within this region were recorded for all of the fluorophores used (depending on the experiment): DAPI, Cy3 (590–650 nm emission) and Cy5 or Alexa 647 (665 nm long pass emission). To automate the analysis ImageJ macros were developed and are available at upon request. For each sample a total of 100 to 500 nuclei were recorded in multiple fields. For cell culture images mH2A and HP1β levels were expressed as average intensities in arbibrary fluorescence units (a.u.). For tissue images mH2A fluorescence intensities were normalized to DAPI intensities for each nucleus. Nuclei were distributed into bins based on their expression levels, and plotted as histograms with the value of each bin shown as % of total nuclei.
Kreiling J.A., Tamamori-Adachi M., Sexton A.N., Jeyapalan J.C., Munoz-Najar U., Peterson A.L., Manivannan J., Rogers E.S., Pchelintsev N.A., Adams P.D, & Sedivy J.M. (2010). Age-associated increase in heterochromatic marks in murine and primate tissues. Aging cell, 10(2), 292-304.
Publication 2010
Antibodies Cell Culture Techniques Cell Nucleus Cells DAPI Equus asinus Fluorescence Medical Devices Microscopy Microscopy, Confocal, Laser Scanning Microscopy, Fluorescence paraform Phosphates Saline Solution Serum Serum Albumin, Bovine Tissue Fixation Tissues Triton X-100

Most recents protocols related to «Tissue Fixation»

1
Histological Evaluation of Implant Integration
Animals were euthanized 30 days after surgery by CO2 asphyxiation. Gross necropsy was performed according to Registry of Industrial Toxicology Animal data (RITA) and North American Control Animal Database (NACAD) guidelines [20 (link)]. Screw positioning and surrounding bone quality were imaged directly following euthanasia using computed tomography imaging (MicroCT: Quantum FX, Perkin Elmer, MA, USA). Implantation sites were then resected en bloc and stored in 4% buffered formaldehyde. To prevent metal oxidation, screws were removed prior to sample decalcification yet after tissue fixation. Thereafter, samples were decalcified (Rapid Decalcifying Solution, Klinipath, Netherlands) and embedded in paraffin. 4 μm sections were stained using hematoxylin (Haemaluin, Fisher Scientific, MA, USA) & eosin (Eosine G(Y), Merck, Germany) stain. Three to five sections were assessed per animal in two locations: at the level of the screw head and in the surgical trajectory. For every animal, two sections containing either the screw hole or trajectory, and visually most extensive damage were scored in a semi-quantitative fashion by a blinded board-certified pathologist using standardized terminology [21 (link)]. A Nikon Eclipse E900 microscope was used for imaging. All slides were screened for relevant tissue processes and cell types to be included for scoring. Next, the range in which these processes took place was determined on an ordinal scale (0 = absent, 1 = mild, 2 = moderate, 3 = severe). Included categories for scoring were: bone damage, fibrosis, presence of histiocytes, inflammation, necrosis, osteoblasts and periosteal reaction. Muscle tissue was separately scored for atrophy, calcifications, fibrosis, necrosis and inflammation. The density of polymorphonuclear cells was used to score inflammation.
Steverink J.G., van Tol F.R., Bruins S., Smorenburg A.J., Tryfonidou M.A., Oosterman B.J., van Dijk M.R., Malda J, & Verlaan J.J. (2023). Lack of concentration-dependent local toxicity of highly concentrated (5%) versus conventional 0.5% bupivacaine following musculoskeletal surgery in a rat model. Journal of Experimental Orthopaedics, 10, 21.
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Publication 2023
Animals Asphyxia Atrophy Autopsy Bones Cells Eosin Euthanasia Fibrosis Formaldehyde Head Hematoxylin Histiocytes Inflammation Metals Microscopy Muscle Tissue Necrosis North American People Operative Surgical Procedures Osteoblasts Ovum Implantation Paraffin Embedding Pathologists Periosteum Physiologic Calcification Stains Tissue Fixation Tissues X-Ray Computed Tomography X-Ray Microtomography
2
Intradermal P. acnes-Induced Acne Model and Treatment
The animal experimental process was approved by the Center for Food and Drug Safety Evaluation, Hubei Center for Disease Control and Prevention, Hubei Academy of Preventive Medicine. The animal experiment ethics approval number was 202110166. All animals were maintained and used in accordance with the Animal Management Rules of the Ministry of Health of the People’s Republic of China and the Guide for the Care and Use of Laboratory Animals—Chinese Version. In addition, all animal experiments were conducted with the assistance of the Wuhan Pinuofei Biological Technology Co. Ltd.
Hemolysis assay test was conducted before the in vivo experiment. Fresh RBCs (4 ml) were separated by centrifuge at 3000 rpm for 10 min at 4°C, washed with saline for three times, and diluted to final concentration [5% (v/v)]. Different samples were separately added to 500 μl of RBCs and incubated at 37°C for 4 hours. At the end of incubation, the solution was centrifuged at 3000 rpm for 10 min at 4°C, and the hemolytic activity was determined at 570 nm with a microplate reader: Rate of hemolysis (%) = [(ODsample − ODnegative)/(ODpositive − ODnegative)] × 100%.
The mice (Balb/c, female, 8 to 10 weeks, and 25 to 30 g) were weighed and anesthetized with the anesthetic isoflurane, the hair on the back of the mice was removed in a sterile biosafety cabinet using a shaving machine and a hair removal cream, and 100 μl of P. acnes (2 × 105 CFUs/ml) was intradermally injected into the skin of the back of the mice after disinfection of the skin with 75% alcohol cotton balls to establish an animal model of common acne. There were five mice per group. The mice were treated with different materials for 7 days. MNs were pressed into the skin by hand against the acne site and held for 3 min. The blank control group, medical acne ointment, patch, blank MN patch, MN patch, and MN patch were treated for 10 min with ultrasound (medical ultrasound probe, 1 cm in diameter). During the treatment period, the swelling volume of acne was measured daily with microcalipers. The changes in the acne on the back of mice were photographed and recorded at 1, 3, and 7 days after the experiment, and after euthanasia treatment, and the skin was cut along 2 mm of the wound edge to complete the tissue acquisition. The expression of factors related to wounds, underlying muscle repair, and inflammatory indexes were analyzed. For skin tissue acquisition and embedding tissue fixation, tissue fixation, tissue dehydration, tissue permeabilization, tissue wax immersion, tissue embedding, tissue sectioning were performed.
Xiang Y., Lu J., Mao C., Zhu Y., Wang C., Wu J., Liu X., Wu S., Kwan K.Y., Cheung K.M, & Yeung K.W. (2023). Ultrasound-triggered interfacial engineering-based microneedle for bacterial infection acne treatment. Science Advances, 9(10), eadf0854.
Publication 2023
Acne Anesthetics Animal Model Animals Animals, Laboratory ARID1A protein, human Biological Assay Biopharmaceuticals Chinese Dehydration Depilation Disinfection Erythrocytes Ethanol Euthanasia Females Food Gossypium Hair Hemolysis Inflammation Isoflurane Mice, Laboratory Muscle Tissue Ointments Safety Saline Solution Skin Sterility, Reproductive Submersion Tissue Fixation Tissues Ultrasonics Ultrasonography Wounds
3
Lung Morphological Assessment
Briefly, immediately after bronchoalveolar lavage, the right lungs were immersed in 4% paraformaldehyde for 24 h. After tissue fixation and paraffin embedding, 5 μm sections were incubated with anti-E-cadherin, ZO-1, Keap1 and Nrf2 antibodies (1:100 dilution) at 4 ℃ overnight followed by incubation with secondary antibody, at last stained with DAB and counterstained with hematoxylin.
Song Y., Fu W., Zhang Y., Huang D., Wu J., Tong S., Zhong M., Cao H, & Wang B. (2023). Azithromycin ameliorated cigarette smoke-induced airway epithelial barrier dysfunction by activating Nrf2/GCL/GSH signaling pathway. Respiratory Research, 24, 69.
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Publication 2023
Antibodies Bronchoalveolar Lavage Cadherins Hematoxylin Immunoglobulins KEAP1 protein, human Lung NFE2L2 protein, human paraform Technique, Dilution Tissue Fixation
4
Tissue Perfusion and Histological Analysis

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Publication 2023
Animals Brain Buffers CM-DiI cresyl violet DAPI Freezing General Anesthesia Heparin Isoflurane Microscopy paraform Phosphates Rattus Sucrose Tissue Fixation
5
Multi-Color Immunofluorescence Staining and Scoring of ESCC
FFPE tissue blocks were cut into 4μm serials slides for hematoxylin & eosin (H&E) staining, immunohistochemistry (IHC) staining and multi-color Immunofluorescence (mIF). For detailed IHC experimental procedures, please refer to our previous publication (30 (link)).
Multi-color immunofluorescence staining and scoring
Five markers CD4, CD8, FOXP3, IL-10 and IgG4 were stained on tissue from surgical samples of 118 ESCC specimens employing TSA-RM-24259 (50T) kits from PANOVUE according to the manufacturer’s instructions. The dilution and fluorescence of antibodies were shown in Supplementary Table 1. The protocols were described briefly below. First, slides were deparaffinized and rehydrated as for conventional IHC. Then, 4% paraformaldehyde was used for tissue re-fixation for 10 min at room temperature followed by antigen retrieval the same as for IHC. After cooled down and washed with 1X TBST 3 times 5min each, slides were further incubated with 10% horse serum for 10 min and then primary antibody for 30min at room temperature. Next, slides were performed with a second antibody for 10 min and PPD fluorescent dye solution, diluted at 1:100 with signal amplifying fluid of the kit, for 10 min. Then, the sections went through antigen retrieval again and entered into next cycle until the 5 markers signals were stained. Finally, the slides were incubated with DAPI and covered with coverslips, and the slides were then ready for multispectral microscopic imaging and automatic cell counting.
Multispectral microscopic imaging employed PerkinElmer Vectra device and automatic counting used inForm® V2.2 advanced image analysis system. More than 5 random high resolutions (200X) microscope fields of tumor parenchyma or TLS were acquired for scoring, which contained tumor and stroma areas or with IgG4 positive cells. Based on computer intelligent recognition algorithm, the software distinguished differently defined areas and identified every single cell according to the DAPI signal of cell nucleus. In the end output, the software displays single positive rates of 5 markers individually and double positive rates of CD4 plus Foxp3. The process of tissue staining and positive cell counting are shown in Figure 1.
Wang H., Su C., Li Z., Ma C., Hong L., Li Z., Ma X., Xu Y., Wei X., Geng Y., Zhang W., Li P, & Gu J. (2023). Evaluation of multiple immune cells and patient outcomes in esophageal squamous cell carcinoma. Frontiers in Immunology, 14, 1091098.
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Publication 2023
Antigens Cell Nucleus Cells DAPI Eosin Equus caballus Fluorescent Antibody Technique Fluorescent Dyes IgG4 IL10 protein, human Immunoglobulins Immunohistochemistry liquid crystal polymer Medical Devices Microscopy Neoplasms Operative Surgical Procedures paraform Serum Technique, Dilution Tissue Fixation Tissues Tissue Stains

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Phosphate-buffered saline (PBS) is a widely used buffer solution in biological research and laboratory procedures. It is a balanced salt solution that maintains a physiological pH and osmolarity, making it suitable for a variety of applications. PBS is primarily used to maintain the viability and integrity of cells, tissues, and other biological samples during various experimental protocols.
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.

More about "Tissue Fixation"

Tissue preservation, histological preparation, cellular stabilization, protein cross-linking, microscopy sample preparation, immunohistochemistry sample preparation, formalin fixation, glutaraldehyde fixation, PubCompare.ai, research protocols, reproducibility, accuracy, literature search, preprint analysis, patent review, research efficiency, NanoZoomer digital slide scanner, DAPI nuclear stain, paraformaldehyde fixative, microtome sectioning, phosphate buffered saline (PBS), laser scanning microscope (LSM 710), epidermal growth factor (EGF), ultra-low attachment tissue culture plates, penicillin/streptomycin antibiotic, MeSH term.
Tissue fixation is a crucial step in many research and diagnostic applications, such as histology, microscopy, and immunohistochemistry.
The fixation process typically involves the use of chemical agents, such as formalin or glutaraldehyde, which cross-link proteins and stabilize cellular components.
Proper tissue fixation is essential for accurate analysis and reproducible results.
Researchers can leverage AI-powered platforms like PubComapre.ai to easily locate, compare, and identify the best tissue fixation protocols from the literature, preprints, and patents, enhancing research effiecy and confindence.