<ol>
	<li>AVMA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> AVMA Guidelines on Euthanasia. , (2007).</li><li>Feldman, D. B., Seely, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Necropsy Guide: Rodents and the rabbit. , (1988).</li><li>King, J. M., Dodd, D. C., Roth, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Necropsy Book. , (2006).</li><li>Kittel, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--Part+2.+A+joint+publication+of+the+RITA+and+NACAD+groups.">Revised guides for organ sampling and trimming in rats and mice--Part 2. A joint publication of the RITA and NACAD groups.</a> Exp Toxicol Pathol. 55, 413-431 (2004).</li><li>Komarek, V., Fox, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+1.">Chapter 1.</a> The Mouse in Biomedical Research. , 1-22 (2007).</li><li>Morawietz, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--Part+3.+A+joint+publication+of+the+RITA+and+NACAD+groups.">Revised guides for organ sampling and trimming in rats and mice--Part 3. A joint publication of the RITA and NACAD groups.</a> Exp Toxicol Pathol. 55, 433-449 (2004).</li><li>Popesko, P., Raijtov&#225;, V., Hor&#225;k, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Colour Atlas of Anatomy of Small Laboratory Animals. 2, 91-106 (2002).</li><li>Ruehl-Fehlert, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--part+1.">Revised guides for organ sampling and trimming in rats and mice--part 1.</a> Exp Toxicol Pathol. 55, 91-106 (2003).</li></ol>

המחברים הם כל העובדים של מכרסמים במעבדה צ&#39;ארלס ריבר של האבחון במעבדה, שבה שירותים אלה מוצעים מסחרית.

איסוף נתונים בסוף המחקר עשוי לדרוש בדיקה שלאחר המוות של בעלי חיים. תאר היטב מה הוא ראה ולזכור לבחון את כל הרקמות. נהלים אלה נועדו בעיקר כדי לייעל necropsy אוסף מדגם לפיקוח מחלות זיהומיות, אך רובם ניתנים להתאמה בקלות החקירות התמקדו התקריות התפרצויות המחלה או חשד בו אולי רק...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><th> שם מגיב </th><th> חברה </th><th> מספר קטלוגי </th><th> תגובות </th></tr><tr><td> קרש חיתוך </td><td> Thermo Electron Corp חתול # 36114 </td><td> חתול # 36114 </td><td></td></tr><tr><td> מספריים קטנים </td><td> ROBOZ </td><td> RS-5910, G204 </td><td> להבי 23mm, 3.5 &quot;אורך, ישר </td></tr><tr><td> בינוני מספריים </td><td> ROBOZ </td><td> RS-6808, G207 </td><td> 5 &quot; </td></tr><tr><td> מספריים גדול </td><td> ROBOZ </td><td> RS-6826, G65 &quot; </td><td> 6.25 &quot; </td></tr><tr><td> מלקחיים-מעוקל </td><td> ROBOZ </td><td> RS-8254 (M1/21004) </td><td> 4.5 &quot;, משונן, עקומה קלה </td></tr><tr><td> מלקחיים-Microdissecting </td><td> ROBOZ </td><td> RS-5238 </td><td> הדסון-(אוולד) </td></tr><tr><td> מלקחיים, רקמות מלקחיים </td><td> ROBOZ </td><td> RS - 8160 </td><td> עכברוש שן </td></tr><tr><td> הלהבה מעקר </td><td> אוקספורד labware מוצרים </td><td> Bacti-Cinerator # 5889-001007 </td><td></td></tr><tr><td> אזמל נתוחים </td><td> אבחון סרטן, Inc </td><td> פינגר אזמלים # 60, # חתול FS0060 </td><td></td></tr><tr><td> Pipettes </td><td> VWR </td><td> פסטר Pipet 5 3 / 4 &quot;, חתול # 14672-400 </td><td></td></tr><tr><td> החיטוי שקיות </td><td> ייצור פרופר החברה </td><td> תיק שקיות נייר מעקר חתול # 021002 (3100923) E09110 </td><td> עבור פסטר pipettes </td></tr><tr><td> פיפטה הנורה </td><td> VWR </td><td> חתול # 56310-240 </td><td></td></tr><tr><td> Eppendorf צינורות </td><td> Sarstedt </td><td> Microtube 2ml SafeSeal, שופט # 72.695 </td><td></td></tr><tr><td> Eppendorf צינורות </td><td> ארגוס </td><td> Microtube Cat 5 מ&quot;ל # T20765-C </td><td></td></tr><tr><td> מזרק </td><td> BD (Beckton דיקנסון) </td><td> 1ml-309602-3 מ&quot;ל 300910 5 מ&quot;ל, 10 מ&quot;ל # 309603 309604 חתול חתול 20mL # 309661 </td><td></td></tr><tr><td> מחטים </td><td> BD </td><td> 26G-18G-309625 305195 </td><td></td></tr><tr><td> ניטרלי שנאגרו פורמלין </td><td> VWR </td><td> 20L NBF 10%, חתול # 16004-128 </td><td></td></tr><tr><td> מלוח </td><td> ThermoScientific </td><td> שנאגרו מלוחים בנק דם, חתול # 23-309-178 </td><td></td></tr><tr><td> Trypticase סויה מרק </td><td> בקטון דיקנסון </td><td> TSB: חתול מיובש, # 211825 </td><td></td></tr><tr><td> פוספט שנאגרו מלוחים </td><td> סיגמא אולדריץ </td><td> HA חוצץ, חתול # P3813-10PAK </td><td></td></tr><tr><td> פורמלין כוסות </td><td> VWR </td><td> חתול 4oz # # 36318-852 8oz 36318-860 16oz חתול חתול # 36318-858 </td><td></td></tr><tr><td> כוסות פורמלין גדול </td><td> Oakridge מוצרים </td><td> 32oz חתול מיכל # 0432-1100 </td><td></td></tr><tr><td> אקסטרה כוס פורמלין גדול </td><td> VWR </td><td> HDPE מיכל 160oz רב קט # 89038-282 </td><td></td></tr><tr><td> תפר חומר </td><td> הנרי שיין </td><td> קלועה משי תפר כירורגי, שופט # 100-5000, M766750 </td><td> לגבי האינפלציה ריאות העכבר </td></tr><tr><td> פתיל </td><td> סטייפלס </td><td> חתול # qua-46173 </td><td> לגבי האינפלציה ריאות חולדה </td></tr></tbody></table>

diagnostic necropsy and selected tissue and sample collection in rats and mice

ישנם סוגים רבים מדגם שעשויים להיות שנאספו חיה מורדמים על מנת לסייע באבחון או לגלות חומרים מזהמים במושבה חיה. איסוף נכון של רקמות לעיבוד היסטולוגית עוד יכול להשפיע על איכות תוצאות הבדיקה. מאמר זה מתאר את התנהלות בדיקה בסיסית ברוטו כולל זיהוי של הלב, הכבד, הריאות, הכליות והטחול, וכן כיצד לאסוף אותם איברים. בנוסף ארבעה רקמה / מדגם טכניקות קשה יותר אוסף הם הפגינו. אוסף ריאות זלוף יכול להיות מאתגר במיוחד כאשר הרקמה צריך להיות מנופח כיאות עם מקבע על מנת בתוך הרקמה לתקן כראוי כדי לאפשר הערכה היסטולוגית יסודית. מאמר זה מדגים את הטכניקה צעד אחר צעד כדי להסיר את הריאות לנפח אותו עם מקבע על מנת להשיג קיבעון אופטימלי של הרקמה בתוך 24 שעות. אוסף המוח יכול להיות מאתגר באותה מידה כמו ברקמה רכה נפגע בקלות. מאמר זה מדגים את הטכניקה צעד אחר צעד כדי לחשוף ולהסיר את המוח מהגולגולת עם נזק מינימלי לרקמות. קשרי לימפה mesenteric הוא סוג מדגם טוב בו כדי לזהות הרבה גורמים מזהמים נפוצים כמו וירוסים enteric להתמיד זמן רב יותר הצומת לימפה מאשר לשפוך בצואה. מאמר זה מדגים את התהליך צעד אחר צעד לאיתור בסביבה נקייה מחיידקים, הסרת בלוטת לימפה mesenteric. לבסוף, זיהוי של חומרים מזהמים של דרכי הנשימה יכולה להתבצע על ידי התרבות חיידקי או בדיקת PCR של האף ו / או הסימפונות נוזל aspirates נלקחה ב necropsy. הליך זה מתאר קבלת והכנת מדגם לשאוב הנשימה לתרבות חיידקי בדיקות PCR.

Watch this Scientific Journal Video about Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice at JoVE.com

Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice

מאמר זה מתאר את ההליכים לביצוע בדיקה שלאחר המוות הבסיסי של עכבר או חולדה, ואת אוסף של איברים בסיסיים, כמו גם סוגי המדגם מאתגר יותר מ להערכה היסטולוגית, מיקרוביולוגית, ו-PCR.

Necropsy לאבחון רקמות אוסף נבחר מדגם של חולדות ועכברים

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal ...

diagnostic-necropsy-selected-tissue-sample-collection-rats

Research

JoVE Journal

Biology

74.0K Views.  Charles River. מאמר זה מתאר את ההליכים לביצוע בדיקה שלאחר המוות הבסיסי של עכבר או חולדה, ואת אוסף של איברים בסיסיים, כמו גם סוגי המדגם מאתגר יותר מ להערכה היסטולוגית, מיקרוביולוגית, ו-PCR.

Video: Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice

Using Micro-Electro-Mechanical Systems (MEMS) to Develop Diagnostic Tools

Using Micro-Electro-Mechanical Systems MEMS to Develop Diagnostic Tools

שימוש Micro-Electro-Mechanical Systems (MEMS) לפתח כלים לאבחון

Manual Restraint and Common Compound Administration Routes in Mice and Rats

Being able to safely and effectively restrain mice and rats is an important part of conducting research. Working confidently and humanely with mice and rats requires a basic competency in handling and restraint methods. This article will present the basic principles required to safely handle animals. One-handed, two-handed, and restraint with specially designed restraint objects will be illustrated. Often, another part of the research or testing use of animals is the effective administration of compounds to mice and rats. Although there are a large number of possible administration routes (limited only by the size and organs of the animal), most are not used regularly in research. This video will illustrate several of the more common routes, including intravenous, intramuscular, subcutaneous, and oral gavage. The goal of this article is to expose a viewer unfamiliar with these techniques to basic restraint and substance administration routes. This video does not replace required hands-on training at your facility, but is meant to augment and supplement that training.

Working safely and humanely with research rodents requires a core competency in handling and restraint methods. This article will present the basic principles required to safely handle and effectively administer compounds to mice and rats.

איפוק ידני ומסלולי מנהל מתחם נפוצים בעכברים וחולדות

Being able to safely and effectively restrain mice and rats is an important part of conducting research.  Working confidently and humanely with mice and ...

Germ Cell Transplantation and Testis Tissue Xenografting in Mice

Germ cell transplantation was developed by Dr. Ralph Brinster and colleagues at the University of Pennsylvania in 19941,2. These ground-breaking studies showed that microinjection of germ cells from fertile donor mice into the seminiferous tubules of infertile recipient mice results in donor-derived spermatogenesis and sperm production by the recipient animal2. The use of donor males carrying the bacterial &beta;-galactosidase gene allowed identification of donor-derived spermatogenesis and transmission of the donor haplotype to the offspring by recipient animals1. Surprisingly, after transplantation into the lumen of the seminiferous tubules, transplanted germ cells were able to move from the luminal compartment to the basement membrane where spermatogonia are located3. It is generally accepted that only SSCs are able to colonize the niche and re-establish spermatogenesis in the recipient testis. Therefore, germ cell transplantation provides a functional approach to study the stem cell niche in the testis and to characterize putative spermatogonial stem cells. To date, germ cell transplantation is used to elucidate basic stem cell biology, to produce transgenic animals through genetic manipulation of germ cells prior to transplantation4,5, to study Sertoli cell-germ cell interaction6,7, SSC homing and colonization3,8, as well as SSC self-renewal and differentiation9,10.
Germ cell transplantation is also feasible in large species11. In these, the main applications are preservation of fertility, dissemination of elite genetics in animal populations, and generation of transgenic animals as the study of spermatogenesis and SSC biology with this technique is logistically more difficult and expensive than in rodents. Transplantation of germ cells from large species into the seminiferous tubules of mice results in colonization of donor cells and spermatogonial expansion, but not in their full differentiation presumably due to incompatibility of the recipient somatic cell compartment with the germ cells from phylogenetically distant species12. An alternative approach is transplantation of germ cells from large species together with their surrounding somatic compartment. We first reported in 2002, that small fragments of testis tissue from immature males transplanted under the dorsal skin of immunodeficient mice are able to survive and undergo full development with the production of fertilization competent sperm13. Since then testis tissue xenografting has been shown to be successful in many species and emerged as a valuable alternative to study testis development and spermatogenesis of large animals in mice14.

Protocols for germ cell transplantation and testis tissue xenografting are described. Germ cell transplantation results in donor-derived spermatogenesis in recipient testes and represents a functional reconstitution assay for identification of spermatogonial stem cells (SSCs). Testis tissue xenografting reproduces testis development and spermatogenesis of various donor species in recipient mice.

נבט השתלת ורקמות testis Xenografting ב עכברים

Germ cell transplantation was developed by Dr. Ralph Brinster and colleagues at the University of Pennsylvania in 19941,2. These ground-breaking studies ...

Ovariectomy and 17&beta;-estradiol Replacement in Rats and Mice: A Visual Demonstration

Estrogens are a family of female sexual hormones with an exceptionally wide spectrum of effects. When rats and mice are used in estrogen research they are commonly ovariectomized in order to ablate the rapidly cycling hormone production, replacing the 17&beta;-estradiol exogenously. There is, however, lack of consensus regarding how the hormone should be administered to obtain physiological serum concentrations. This is crucial since the 17&beta;-estradiol level/administration method profoundly influences the experimental results1-3. We have in a series of studies characterized the different modes of 17&beta;-estradiol administration, finding that subcutaneous silastic capsules and per-oral nut-cream Nutella are superior to commercially available slow-release pellets (produced by the company Innovative Research of America) and daily injections in terms of producing physiological serum concentrations of 17&beta;-estradiol4-6. Amongst the advantages of the nut-cream method, that previously has been used for buprenorphine administration7, is that when used for estrogen administration it resembles peroral hormone replacement therapy and is non-invasive. The subcutaneous silastic capsules are convenient and produce the most stable serum concentrations. This video article contains step-by-step demonstrations of ovariectomy and 17&beta;-estradiol hormone replacement by silastic capsules and peroral Nutella in rats and mice, followed by a discussion of important aspects of the administration procedures.

Ovariectomy and 17&amp;#946;-estradiol Replacement in Rats and Mice: A Visual Demonstration

Ovariectomy and subsequent replacement of 17&beta;-estradiol by means of silastic capsules and peroral Nutella are demonstrated in rats and mice.

Ovariectomy ו-17β אסטרדיול החלפת בחולדות ועכברים: הפגנה חזותית

Estrogens are a family of female sexual hormones with an exceptionally wide spectrum of effects. When rats and mice are used in estrogen research they are ...

Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue

Preserved clinical material is a unique source for proteomic investigation of human disorders. Here we describe an optimized protocol allowing large scale quantitative analysis of formalin fixed and paraffin embedded (FFPE) tissue. The procedure comprises four distinct steps. The first one is the preparation of sections from the FFPE material and microdissection of cells of interest. In the second step the isolated cells are lysed and processed using 'filter aided sample preparation' (FASP) technique. In this step, proteins are depleted from reagents used for the sample lysis and are digested in two-steps using endoproteinase LysC and trypsin. 
After each digestion, the peptides are collected in separate fractions and their content is determined using a highly sensitive fluorescence measurement. Finally, the peptides are fractionated on 'pipette-tip' microcolumns. The LysC-peptides are separated into 4 fractions whereas the tryptic peptides are separated into 2 fractions. In this way prepared samples allow analysis of proteomes from minute amounts of material to a depth of 10,000 proteins. Thus, the described workflow is a powerful technique for studying diseases in a system-wide-fashion as well as for identification of potential biomarkers and drug targets.

Archival formalin fixed and paraffin embedded (FFPE) clinical samples are valuable material for investigation of diseases. Here we demonstrate a sample preparation workflow allowing in-depth proteomic analysis of microdissected FFPE tissue.

לדוגמא הכנת proteomic מפורמלין קבועה ורקמות פרפין Embedded

Preserved clinical material is a unique source for proteomic investigation of human disorders. Here we describe an optimized protocol allowing large scale ...

Milk Collection Methods for Mice and Reeves' Muntjac Deer

Animal models are commonly used throughout research laboratories to accomplish what would normally be considered impractical in a pathogen&#8217;s native host. Milk collection from animals allows scientists the opportunity to study many aspects of reproduction including vertical transmission, passive immunity, mammary gland biology, and lactation. Obtaining adequate volumes of milk for these studies is a challenging task, especially from small animal models. Here we illustrate an inexpensive and facile method for milk collection in mice and Reeves&#8217; muntjac deer that does not require specialized equipment or extensive training. This particular method requires two researchers: one to express the milk and to stabilize the animal, and one to collect the milk in an appropriate container from either a Muntjac or mouse model. The mouse model also requires the use of a P-200 pipetman and corresponding pipette tips. While this method is low cost and relatively easy to perform, researchers should be advised that anesthetizing the animal is required for optimal milk collection.

Milk Collection Methods for Mice and Reeves&#039; Muntjac Deer

Milk collection from animal models facilitates various research avenues: understanding passive immunity, identifying pathogens responsible for vertical transmission and, through the use of transgenic mice, even commercial production of proteins found in human breast milk. Here we illustrate a simple method for milk collection in mice and Reeves&#8217; muntjac deer.

שיטות איסוף חלב לעכברים וריבס &#39;Muntjac צביים

Animal models are commonly used throughout research laboratories to accomplish what would normally be considered impractical in a pathogen&#8217;s native ...

Dissection of the Auditory Bulla in Postnatal Mice: Isolation of the Middle Ear Bones and Histological Analysis

In most mammals, auditory ossicles in the middle ear, including the malleus, incus and stapes, are the smallest bones. In mice, a bony structure called the auditory bulla houses the ossicles, whereas the auditory capsule encloses the inner ear, namely the cochlea and semicircular canals. Murine ossicles are essential for hearing and thus of great interest to researchers in the field of otolaryngology, but their metabolism, development, and evolution are highly relevant to other fields. Altered bone metabolism can affect hearing function in adult mice, and various gene-deficient mice show changes in morphogenesis of auditory ossicles in utero. Although murine auditory ossicles are tiny, their manipulation is feasible if one understands their anatomical orientation and 3D structure. Here, we describe how to dissect the auditory bulla and capsule of postnatal mice and then isolate individual ossicles by removing part of the bulla. We also discuss how to embed the bulla and capsule in different orientations to generate paraffin or frozen sections suitable for preparation of longitudinal, horizontal, or frontal sections of the malleus. Finally, we enumerate anatomical differences between mouse and human auditory ossicles. These methods would be useful in analyzing pathological, developmental and evolutionary aspects of auditory ossicles and the middle ear in mice.

We present a protocol to isolate the auditory bulla, capsule, and ossicles from postnatal mice for whole mount and histological analysis.

דיסקציה של בולה השמיעה של עכברים אחרי לידה: בידוד של עצמות באוזן התיכונה ניתוח היסטולוגית

In most mammals, auditory ossicles in the middle ear, including the malleus, incus and stapes, are the smallest bones. In mice, a bony structure called ...

Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually performed as terminal analysis under urethane anesthesia. It is well known that anesthetic drugs can influence bladder function. Hence, the aim of this technique is to perform cystometric measurements of the urinary bladder and external urethral sphincter in lightly restrained awake rats. For this purpose, a bladder catheter is implanted into the bladder dome. Subsequently, two electrodes are implanted bilateral to the external urethral sphincter and a ground electrode is sutured to a non-responsive skeletal muscle. The bladder catheter and the three electrodes are finally tunneled subcutaneously to the neck region and affixed to a harness. With this technique, the lower urinary tract can be measured at multiple time points in the same animal to assess lower urinary tract function. The main application of this technique is the follow-up of simultaneous urinary bladder and external urethral sphincter function in awake healthy rats and after induction of a disease or injury. Moreover, subsequent lower urinary tract monitoring can be performed during evaluation of the disease/injury and to monitor treatment efficacy.

This protocol first describes the surgical procedure of the permanent implantation of a urinary bladder catheter combined with external urethral sphincter electrodes, and second, the measurement of the function of the urinary bladder and external urethral sphincter in implanted awake animals.

מדידות סוגר השופכה Cystometric או חיצוני בחולדות ער עם קטטר מושתל ואלקטרודות ומאפשר למדידות חוזרות

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually ...

Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also termed the hypothalamic-pituitary-gonadal axis. The reproductive axis is a key integrator of environmental and internal input adapting fertility to favorable conditions for reproduction. Prior to embarking upon a fertility study in mice and rats, sexual maturity is evaluated to exclude the possibility that the observed reproductive phenotypes are caused by delayed or absent pubertal onset. This protocol describes a non-invasive approach to assess pubertal onset in males through the determination of preputial separation, and in females through vaginal opening and first estrus. After the confirmation of the completion of puberty and the achievement of sexual maturity, a fertility study can be initiated. The procedure describes the optimal breeding conditions for mice and rats, how to set up a fertility study, and what parameters to evaluate and determine if the treatment or gene deletion has an impact on fertility.

Many treatments and genetic mutations impact the timing of sexual maturity and fertility. This protocol describes a non-invasive method to evaluate pubertal onset in mice and rats prior to setting up a fertility study in sexually mature animals.

קביעת כשירות הרבייה על-ידי המאשר תחילת המתבגר וביצוע וזמינותו של פוריות ב עכברים וחולדות

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also ...

Tissue Collection of Bats for -Omics Analyses and Primary Cell Culture

As high-throughput sequencing technologies advance, standardized methods for high quality tissue acquisition and preservation allow for the extension of these methods to non-model organisms. A series of protocols to optimize tissue collection from bats has been developed for a series of high-throughput sequencing approaches. Outlined here are protocols for the capture of bats, desired demographics to be collected for each bat, and optimized methods to minimize stress on a bat during tissue collection. Specifically outlined are methods for collecting and treating tissue to obtain (i) DNA for high molecular weight genomic analyses, (ii) RNA for tissue-specific transcriptomes, and (iii) proteins for proteomic-level analyses. Lastly, also outlined is a method to avoid lethal sampling by creating viable primary cell cultures from wing clips. A central motivation of these methods is to maximize the amount of potential molecular and morphological data for each bat and suggest optimal ways to preserve tissues so they retain their value as new methods develop in the future. This standardization has become particularly important as initiatives to sequence chromosome-level, error-free genomes of species across the world have emerged, in which multiple scientific parties are spearheading the sequencing of different taxonomic groups. The protocols outlined herein define the ideal tissue collection and tissue preservation methods for Bat1K, the consortium that is sequencing the genomes of every species of bat.

This is a protocol for the optimal tissue preparation for genomic, transcriptomic, and proteomic analyses of bats caught in the wild. It includes protocols for bat capture and dissection, tissue preservation, and cell culturing of bat tissue.

אוסף רקמות של עטלפים עבור-ניתוח Omics ותרבות התא הראשוני

As high-throughput sequencing technologies advance, standardized methods for high quality tissue acquisition and preservation allow for the extension of ...