<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הכן עובש מהקלטת עבור פלטפורמת אוקטובר </li><li style=";text-align:right;direction:rtl"> מלא עובש עם אוקטובר להקפיא בתוך cryostat או באמצעות קרח יבש כתוש. </li><li style=";text-align:right;direction:rtl"> הסר את הסרט מרחבי פלטפורמה אוקטובר קפוא. </li><li style=";text-align:right;direction:rtl"> יישר סימנים על צ&#39;אק מקפיא בשלב cryostat גובר לנעול צ&#39;אק. </li><li style=";text-align:right;direction:rtl"> סעיף באמצעות פלטפורמת אוקטובר עד משטח שטוח. </li><li style=";text-align:right;direction:rtl"> הסר resurfaced פלטפורמה אוקטובר מקום על הבמה הקפאת cryostat. </li><li style=";text-align:right;direction:rtl"> דגימת רקמה פלייס (cryopreserved בעבר עם גליצרול 30% או סוכרוז ב PBS) ב אוקטובר </li><li style=";text-align:right;direction:rtl"> הכן את העמודה הקפאה עם הטבעת החיצונית מקרין על 5 מ&quot;מ מעל גבי טור להרכיב גם עבור אוקטובר </li><li style=";text-align:right;direction:rtl"> בזהירות עמדה דגימת רקמה על פני השטח של מרכז עמודה מקפיא לאט להוסיף עד אוקטובר היטב מלא. </li><li style=";text-align:right;direction:rtl"> הקף טור מקפיא עם קרח יבש כתוש. רקמות אוקטובר צריך להקפיא לחלוטין בתוך 20-60 שניות. </li><li style=";text-align:right;direction:rtl"> עם העלייה בטמפרטורה הכנה, הטבעת החיצונית ניתן להסיר תוך המדגם נשאר קפוא. </li><li style=";text-align:right;direction:rtl"> שקופיות מדגם הצידה טור מקפיא מקום cryostat. </li><li style=";text-align:right;direction:rtl"> מניחים ירידה של אוקטובר על פני השטח של פלטפורמת אוקטובר ואת הדגימה עמדה (רקמה למטה) הפעלת לחץ תקיף. הדגימה במהרה להקפיא על גבי פלטפורמת אוקטובר </li><li style=";text-align:right;direction:rtl"> צ&#39;אק מאובטחת על cryostat הרכבה הבמה עם סימני מיושר. </li><li style=";text-align:right;direction:rtl"> סעיף דרך אוקטובר שטחית כדי הדגימה רקמות. </li><li style=";text-align:right;direction:rtl"> הפשירי הר חלקים דקים על גבי שקופיות זכוכית ולאחסן קפוא או בטמפרטורת החדר. </li><li style=";text-align:right;direction:rtl"> Immunoreactions יכול להתבצע עבור רקמה רכוב על שקופיות הזכוכית. </li><li style=";text-align:right;direction:rtl"> הוא מגיב ונקווה לשקופית, יכול להיות נסער בעדינות, עשויים להיות מכוסים אם רגישים לאור. </li><li style=";text-align:right;direction:rtl"> בשלבים מאוחרים יותר של התגובה מבוצעים בקלות לתוך כלי קיבול אשפה על ידי היפוך שקופיות, הפתילה את השקופית, ולאחר מכן להחיל את מגיב הבאה. </li></ol>

<ol>
	<li>Scoutern, C. W., O'Connor, R., Cunningham, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion+fixation+of+research+animals.">Perfusion fixation of research animals.</a> Microsc. Today. 14, 26-33 (2006).</li><li>Cunningham, M. G., Connor, C. M., Zhang, K., Benes, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diminished+serotonergic+innervation+of+adult+medial+prefrontal+cortex+after+6-OHDA+lesions+in+the+newborn+rat.">Diminished serotonergic innervation of adult medial prefrontal cortex after 6-OHDA lesions in the newborn rat.</a> Brain Res. Dev. Brain Res. 157, 124-131 (2005).</li></ol>

הפרוטוקול המובא כאן מספק החוקרים עם תמציתי, קל לעקוב אחר קווי המתאר של כיצד להשיג חלקים cryostat דקה, קשה לניהול, חתיכות קטנות רקמות, כגון ביופסיה, פרוסות המוח למחקרים נוספים להתבצע, כגון מכתים שיטות שונות, הכלאה באתרו, או אימונוהיסטוכימיה.

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<td>O. C. T.</td>
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thin sectioning of slice preparations for immunohistochemistry

חקירות רבות במדעי המוח, כמו גם מתחומים אחרים, לערב לימוד קטנות, אך חלקים מאקרוסקופיים או חלקים של רקמת שנשתמרו, הסיר טרי, או נכרת, אבל המשיכה קיימא, כמו פרוסת ההכנות של רקמת המוח. מחקרים מאוחרים יותר מיקרוסקופיים של חומר זה יכול להיות מאתגר, כמו דגימות רקמה עלול להיות קשה להתמודד. הפגינו כאן היא שיטה לקבלת סעיפים cryostat דקה של רקמה בעובי זה עשוי לנוע בין 0.2-5.0 מ&quot;מ. אנו שגרתי לחתוך עבה 400 מיקרון Vibratome פרוסות המוח סדרתי לתוך סעיפים 5-10 מיקרון העטרה cryostat. פרוסות הם בדרך כלל בשימוש הראשון של הניסויים electrophysiology ואז דורשים ניתוח מיקרוסקופי של cytoarchitecture של האזור שממנו הקלטות נצפו. אנחנו צריכים לבנות מכשיר פשוט המאפשר הכנה מבוקרת לשחזור והמיקום של הפרוסה רקמות. מכשיר זה מורכב 5 ס&quot;מ אורך גליל בקוטר של 1.2 ס&quot;מ, המשמש כבמה הקפאת עבור הפרוסה. טבעת הדוקה שקופיות על קירות מתן גליל סביב פרוסה המאפשר את הרקמה להיות שקוע בתוך מתחם הקפאה (למשל, אוקטובר). זהו אז קפוא במהירות עם קרח יבש כתוש רקיק וכתוצאה מכך יכול להיות בעמדה בקלות חתך cryostat. סעיפים Thin ניתן להפשיר רכוב על גבי שקופיות מצופה לאפשר מחקרים נוספים שיש לבצע, כגון שיטות צביעה שונים, הכלאה באתרו, או אימונוהיסטוכימיה, כפי שהודגם כאן.

Watch this Scientific Journal Video about Thin Sectioning of Slice Preparations for Immunohistochemistry at JoVE.com

Thin Sectioning of Slice Preparations for Immunohistochemistry

השיטה הנוכחית מאפשרת cryostat לשחזור של חתך, קשה לניהול, חתיכות קטנות רקמות, כגון ביופסיה, פרוסות המוח. אנו מנצלים אלומיניום פשוט מקפיא הבמה כדי להקל על הטיפול רקמות cryostat רגיל שגרתי לייצר חלקים 5-10 מיקרון סדרתי של פרוסות 400 מיקרון עבה המוח.

חתך דק של ההכנות פורסים עבור אימונוהיסטוכימיה

Many investigations in neuroscience, as well as other disciplines, involve studying small, yet macroscopic pieces or sections of tissue that have been ...

thin-sectioning-of-slice-preparations-for-immunohistochemistry

Research

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Biology

32.7K Views.  Yonsei University College of Medicine, Severance Hospital. השיטה הנוכחית מאפשרת cryostat לשחזור של חתך, קשה לניהול, חתיכות קטנות רקמות, כגון ביופסיה, פרוסות המוח. אנו מנצלים אלומיניום פשוט מקפיא הבמה כדי להקל על הטיפול רקמות cryostat רגיל שגרתי לייצר חלקים 5-10 מיקרון סדרתי של פרוסות 400 מיקרון עבה המוח.

Video: Thin Sectioning of Slice Preparations for Immunohistochemistry

Organotypic Slice Culture of E18 Rat Brains

Organotypic slice cultures from embryonic rodent brains are widely used to study brain development. While there are often advantages to an in-vivo system, organotypic slice cultures allow one to perform a number of manipulations that are not presently feasible in-vivo. To date, organtotypic embryonic brain slice cultures have been used to follow individual cells using time-lapse microscopy, manipulate the expression of genes in the ganglionic emanances (a region that is hard to target by in-utero electroporation), as well as for pharmacological studies. In this video protocol we demonstrate how to make organotypic slice cultures from rat embryonic day 18 embryos. The protocol involves dissecting the embryos, embedding them on ice in low melt agarose, slicing the embedded brains on the vibratome, and finally plating the slices onto filters in culture dishes. This protocol is also applicable in its present form to making organotypic slice cultures from different embryonic ages for both rats and mice. 

פורסים תרבות Organotypic של E18 מוח החולדה

Organotypic slice cultures from embryonic rodent brains are widely used to study brain development. While there are often advantages to an in-vivo system, ...

Preparation of Gene Gun Bullets and Biolistic Transfection of Neurons in Slice Culture

Biolistic transfection is a physical means of transfecting cells by bombarding tissue with high velocity DNA coated particles. We provide a detailed protocol for biolistic transfection of rat hippocampal slices, from the initial preparation of DNA coated bullets to the final shooting of the organotypic slice cultures using a gene gun. Gene gun transfection is an efficient and easy means of transfecting neurons and is especially useful for fluorescently labeling a small subset of cells in tissue slice. In this video, we first outline the steps required to coat gold particles with DNA. We next demonstrate how to line the inside of plastic tubing with the gold/DNA bullets, and how to cut this tubing to obtain the plastic cartridges for loading into the gene gun. Finally, we perform biolistic transfection of rat hippocampal slice cultures, demonstrating handling of the Bio-Rad Helios gene gun, and offering trouble shooting advice to obtain healthy and optimally transfected tissue slices.  

We describe a method for preparing DNA coated gold bullets and demonstrate the use of such bullets to biolistically transfect neurons in cultured hippocampal slices.

הכנת כדורי אקדח ין transfection Biolistic של נוירונים תרבות Slice

Biolistic transfection is a physical means of transfecting cells by bombarding tissue with high velocity DNA coated particles. We provide a detailed ...

Drosophila Larval NMJ Immunohistochemistry

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use of the Drosophila motor system is due to its high accessibility. It can be analyzed with single-cell resolution. There are 30 muscles per hemisegment whose arrangement within the peripheral body wall are known. A total of 31 motor neurons attach to these muscles in a pattern that has high fidelity. Using molecular biology and genetics, one can create transgenic animals or mutants. Then, one can study the developmental consequences on the morphology and function of the NMJ. Immunohistochemistry can be used to clearly image the components of the NMJ. In this article, we demonstrate how to use antibody staining to visualize the Drosophila larval NMJ.

Drosophila Larval NMJ Immunohistochemistry

This protocol demonstrates how to perform immunohistochemistry on dissected Drosophila larva.

תסיסנית הזחל NMJ אימונוהיסטוכימיה

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use ...

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis

Drosophila has long been used as model system to study development, mainly due to the ease with which it is genetically tractable. Over the years, a plethora of mutant strains and technical tricks have been developed to allow sophisticated questions to be asked and answered in a reasonable amount of time. Fundamental insight into the interplay of components of all known major signaling pathways has been obtained in forward and reverse genetic Drosophila studies. The fly eye has proven to be exceptionally well suited for mutational analysis, since, under laboratory conditions, flies can survive without functional eyes. Furthermore, the surface of the insect eye is composed of some 800 individual unit eyes (facets or ommatidia) that form a regular, smooth surface when looked at under a dissecting microscope. Thus, it is easy to see whether a mutation might affect eye development or growth by externally looking for the loss of the smooth surface ('rough eye' phenotype; Fig. 1) or overall eye size, respectively (for examples of screens based on external eye morphology see e.g.1). Subsequent detailed analyses of eye phenotypes require fixation, plastic embedding and thin-sectioning of adult eyes.
The Drosophila eye develops from the so-called eye imaginal disc, a bag of epithelial cells that proliferate and differentiate during larval and pupal stages (for review see e.g. 2). Each ommatidium consists of 20 cells, including eight photoreceptors (PR or R-cells; Fig. 2), four lens-secreting cone cells, pigment cells ('hexagon' around R-cell cluster) and a bristle. The photoreceptors of each ommatidium, most easily identified by their light sensitive organelles, the rhabdomeres, are organized in a trapezoid made up of the six "outer" (R1-6) and two "inner" photoreceptors (R7/8; R8 [Fig. 2] is underneath R7 and thus only seen in sections from deeper areas of the eye). The trapezoid of each facet is precisely aligned with those of its neighbors and the overall anteroposterior and dorsoventral axes of the eye (Fig. 3A). In particular, the ommatidia of the dorsal and ventral (black and red arrows, respectively) halves of the eye are mirror images of each other and correspond to two chiral forms established during planar cell polarity signaling (for review see e.g. 3).
The method to generate semi-thin eye sections (such as those presented in Fig. 3) described here is slightly modified from the one originally described by Tomlinson and Ready4. It allows the morphological analysis of all cells except for the transparent cone cells. In addition, the pigment of R-cells (blue arrowheads in Fig. 2 and 3) can be used as a cell-autonomous marker for the genotype of a R-cell, thus genetic requirements of genes in a subset of R-cells can readily be determined5,6.

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis

A standard approach to prepare adult Drosophila eyes for semi-thin sectioning and light microscopic analysis is presented here. The protocol can be used for gross morphological analysis of eye defects, or with the indicated adjustments can be used to determine genetic requirements of genes in specific cell types of the eye (e.g. clonal analysis of photoreceptors) or for electron microscopic analysis.

הכנת למבוגרים תסיסנית עיניים עבור חתך דק ניתוח מיקרוסקופי

Drosophila has long been used as model system to study development, mainly due to the ease with which it is genetically tractable. Over the years, a ...

Drosophila Pupal Abdomen Immunohistochemistry

The Drosophila pupal abdomen is an established model system for the study of epithelial morphogenesis and the development of sexually dimorphic morphologies 1-3. During pupation, which spans approximately 96 hours (at 25 &deg;C), proliferating populations of imaginal cells replace the larval epidermis to generate the adult abdominal segments. These imaginal cells, born during embryogenesis, exist as lateral pairs of histoblast nests in each abdominal segment of the larvae. Four pairs of histoblast nests give rise to the adult dorsal cuticle (anterior and posterior dorsal nests), the ventral cuticle (ventral nests) and the spiracles associated with each segment (spiracle nests) 4. Upon puparation, these diploid cells (distinguishable by size from the larger polyploid larval epidermal cells- LECs) begin a stereotypical process of proliferation, migration and replacement of the LECs. Various molecular and genetic tools can be employed to investigate the contributions of genetic pathways involved in morphogenesis of the adult abdomen. Ultimate adult phenotypes are typically analyzed following dissection of adult abdominal cuticles. However, investigation of the underlying molecular processes requires immunohistochemical analyses of the pupal epithelium, which present unique challenges. Temporally dynamic morphogenesis and the interactions of two distinct epithelial populations (larval and imaginal) generate a fragile tissue prone to excessive cell loss during dissection and subsequent processing. We have developed methods of dissection, fixation, mounting and imaging of the Drosophila pupal abdominem epithelium for immunohistochemical studies that generate consistent high quality samples suitable for confocal or standard fluorescent microscopy.

Drosophila Pupal Abdomen Immunohistochemistry

Antibody staining of the Drosophila pupae can enhance genetic analyses of adult abdominal developmental genetics. We present our protocol for dissection, fixation and antibody staining of staged Drosophila pupal abdomen.

תסיסנית גלמי אימונוהיסטוכימיה בטן

The Drosophila pupal abdomen is an established model system for the study of epithelial morphogenesis and the development of sexually dimorphic ...

Production of Tissue Microarrays, Immunohistochemistry Staining and Digitalization Within the Human Protein Atlas

The tissue microarray (TMA) technology provides the means for high-throughput analysis of multiple tissues and cells. The technique is used within the Human Protein Atlas project for global analysis of protein expression patterns in normal human tissues, cancer and cell lines. Here we present the assembly of 1 mm cores, retrieved from microscopically selected representative tissues, into a single recipient TMA block. The number and size of cores in a TMA block can be varied from approximately forty 2 mm cores to hundreds of 0.6 mm cores. The advantage of using TMA technology is that large amount of data can rapidly be obtained using a single immunostaining protocol to avoid experimental variability. Importantly, only limited amount of scarce tissue is needed, which allows for the analysis of large patient cohorts 1 2. Approximately 250 consecutive sections (4 &mu;m thick) can be cut from a TMA block and used for immunohistochemical staining to determine specific protein expression patterns for 250 different antibodies. In the Human Protein Atlas project, antibodies are generated towards all human proteins and used to acquire corresponding protein profiles in both normal human tissues from 144 individuals and cancer tissues from 216 different patients, representing the 20 most common forms of human cancer. Immunohistochemically stained TMA sections on glass slides are scanned to create high-resolution images from which pathologists can interpret and annotate the outcome of immunohistochemistry. Images together with corresponding pathology-based annotation data are made publically available for the research community through the Human Protein Atlas portal (<a href="http://www.proteinatlas.org" target="_blank">www.proteinatlas.org</a>) (Figure 1) 3 4. The Human Protein Atlas provides a map showing the distribution and relative abundance of proteins in the human body. The current version contains over 11 million images with protein expression data for 12.238 unique proteins, corresponding to more than 61% of all proteins encoded by the human genome.

Tissue microarrays allows for an efficient method to gain concurrent information from a multitude of tissues. Representative parts of tissues are assembled into a single paraffin block. Sections from the block are used for immunohistochemistry and analysis of protein expression patterns. Digital scanning generates corresponding images for distribution of data.

הפקה של microarrays רקמות, צביעה אימונוהיסטוכימיה ו דיגיטציה בתוך אטלס החלבון האנושי

The  tissue microarray (TMA) technology provides the means for high-throughput  analysis of multiple tissues and cells. The technique is used within the ...

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ring-supported hydrophilized PTFE membrane to provide structural support to both living and fixed tissue during immunohistochemical processing, which allows for the use of a variety of protocols that would otherwise cause damage to the tissue. First, this is demonstrated with bolus loading of fluorescent markers into living retinal tissue. This method allows for quick visualization of targeted structures, while the membrane support maintains tissue integrity during the injection and allows for easy transfer of the preparation for further imaging or processing.
Second, a procedure for antibody staining in tissue fixed with carbodiimide is described. Though paraformaldehyde fixation is more common, carbodiimide fixation provides a superior substrate for the visualization of synaptic proteins. A limitation of carbodiimide is that the resulting fixed tissue is relatively fragile; however, this is overcome with the use of the supporting membrane. Retinal tissue is used to demonstrate these techniques, but they may be applied to any fragile tissue.

The protocol described here is for structural assessment of a wholemount retinal preparation. This includes descriptions of tissue dissection, mounting onto a hydrophilized polytetrafluoroethylene (PTFE) membrane insert, bolus loading with fluorescent markers, and a comparison of fixation with carbodiimide and paraformaldehyde for immunohistochemical analysis of cellular and synaptic components.

פרוטוקול אופטימלי עבור רשתית Wholemount הכנה להדמיה ואימונוהיסטוכימיה

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ...

Spatiotemporal Mapping of Motility in Ex Vivo Preparations of the Intestines

Multiple approaches have been used to record and evaluate gastrointestinal motility including: recording changes in muscle tension, intraluminal pressure, and membrane potential. All of these approaches depend on measurement of activity at one or multiple locations along the gut simultaneously which are then interpreted to provide a sense of overall motility patterns. Recently, the development of video recording and spatiotemporal mapping (STmap) techniques have made it possible to observe and analyze complex patterns in ex vivo whole segments of colon and intestine. Once recorded and digitized, video records can be converted to STmaps in which the luminal diameter is converted to grayscale or color [called diameter maps (Dmaps)]. STmaps can provide data on motility direction (i.e., stationary, peristaltic, antiperistaltic), velocity, duration, frequency and strength of contractile motility patterns. Advantages of this approach include: analysis of interaction or simultaneous development of different motility patterns in different regions of the same segment, visualization of motility pattern changes over time, and analysis of how activity in one region influences activity in another region. Video recordings can be replayed with different timescales and analysis parameters so that separate STmaps and motility patterns can be analyzed in more detail. This protocol specifically details the effects of intraluminal fluid distension and intraluminal stimuli that affect motility generation. The use of luminal receptor agonists and antagonists provides mechanistic information on how specific patterns are initiated and how one pattern can be converted into another pattern. The technique is limited by the ability to only measure motility that causes changes in luminal diameter, without providing data on intraluminal pressure changes or muscle tension, and by the generation of artifacts based upon experimental setup; although, analysis methods can account for these issues. When compared to previous techniques the video recording and STmap approach provides a more comprehensive understanding of gastrointestinal motility.

Spatiotemporal Mapping of Motility in Ex Vivo Preparations of the Intestines

Recently available video recording and spatiotemporal mapping (STmap) techniques make it possible to visualize and quantify both propagating and mixing patterns of intestinal motility. The goal of this protocol is to explain the generation and analysis of STmaps using the GastroIntestinal Motility Monitoring (GIMM) system.

מיפוי Spatiotemporal של תנועתיות ב<em&gt; Ex Vivo</em&gt; תכשירים מהמעיים

Multiple approaches have been used to record and evaluate gastrointestinal motility including: recording changes in muscle tension, intraluminal pressure, ...

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. 5-hydroxylmethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC) are the recently discovered modified cytosine bases produced by enzymatic oxidation of 5mC, whose biological functions remain relatively obscure. A number of approaches ranging from biochemical to antibody based techniques have been employed to study the genomic distribution and global content of these modifications in various biological systems. Although some of these approaches can be useful for quantitative assessment of these modified forms of 5mC, most of these methods do not provide any spatial information regarding the distribution of these DNA modifications in different cell types, required for correct understanding of their functional roles. Here we present a highly sensitive method for immunochemical detection of the modified forms of cytosine. This method permits co-detection of these epigenetic marks with protein lineage markers and can be employed to study their nuclear localization, thus, contributing to deciphering their potential biological roles in different experimental contexts.

Herein we describe a sensitive immunochemical method for mapping the spatial distribution of 5mC oxidation derivatives based on the use of peroxidase-conjugated secondary antibodies and tyramide signal amplification.

איתור של טפסים השתנו ציטוזין שימוש אימונוהיסטוכימיה רגיש

Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. ...

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples

Maintaining plant section integrity is essential for studying detailed anatomical structures at the cellular, tissue, or even organ level. However, some plant cells have rigid cell walls, tough fibers and crystals (calcium oxalate, silica, etc.), and high water content that often disrupt tissue integrity during plant tissue sectioning.
This study establishes a simple Hybrid-Cut tissue sectioning method. This protocol modifies a paraffin-based sectioning technique and improves the integrity of tissue sections from different plants. Plant tissues were embedded in paraffin before sectioning in a cryostat at -16 &#176;C. Sectioning under low temperature hardened the paraffin blocks, reduced tearing and scratching, and improved tissue integrity significantly. This protocol was successfully applied to calcium oxalate-rich Phalaenopsis orchid tissues as well as recalcitrant tissues such as reproductive organs and leaves of rice, maize, and wheat. In addition, the high quality of tissue sections from Hybrid-Cut could be used in combination with in situ hybridization (ISH) to provide spatial expression patterns of genes of interest. In conclusion, this protocol is particularly useful for recalcitrant plant tissue containing high crystal or silica content. Good quality tissue sections enable morphological and other biological studies.

This protocol describes a simple Hybrid-Cut tissue sectioning method that is useful for recalcitrant plant tissues. Good quality tissue sections enable anatomical studies and other biological studies including in situ hybridization (ISH).

-Cut ההיברידי: שיטת חתך משופרת עבור דוגמאות רקמות צמח סרבן

Maintaining plant section integrity is essential for studying detailed anatomical structures at the cellular, tissue, or even organ level. However, some ...