ברצוננו להודות לקרן קנדית עבור חדשנות, פיתוח קולומביה הבריטית קרן ידע מדעי הלאומי וההנדסה מועצת המחקר (NSERC) של קנדה לתמיכה מחקרים מתמשכים על אזורים חמצן נמוכה של מים חופי האוקיינוס ​​הפתוח. בנוסף, דאו נתמכה על ידי מלגות של NSERC, Killam ויסוד טולה מרכז מימנה עבור גיוון התפתחות חיידקים.

חלק 1: איסוף מים <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ממעמקים מוקצים להפיל כל אחד. </li><li style=";text-align:right;direction:rtl"> ממעמקים להיות שנדגמו ואת הנפח הכולל להיות שנאספו עבור השחקנים הנוכחי נחושים. </li><li style=";text-align:right;direction:rtl"> היא הבטיחה כי Go-פלו ובקבוקים Niskin פתוחים, וכי השסתומים שלהם הברזים סגורים. </li><li style=";text-align:right;direction:rtl"> CTD הוא הועמסו על התחתונה השורה התחתונה לתוך המים. </li><li style=";text-align:right;direction:rtl"> בקבוק עבור עומק העמוק ביותר שנדגמו בגבס הראשון הוא הועמסו על הקו לאחר CTD, היא הבטיחה כי קו מצויד כהלכה לתוך סוגר את הברגים הם הידקו. </li><li style=";text-align:right;direction:rtl"> שליח מצורף בקבוק אחד (למעט אחד העמוקים ביותר) לפני הורדת. </li><li style=";text-align:right;direction:rtl"> הקו הוא הוריד את המרחק בין עומק הראשונה הבאה; עוד בקבוק נטען אז. יצוין כי שם במעמקי 20L נאספים לסינון, סך של 2 בקבוקים 12L נטענים ברציפות על 0.75 מ &#39;זה מזה. </li><li style=";text-align:right;direction:rtl"> לאחר כל הבקבוקים הם על הקו, קו יורד לעומק הרצוי 2 דקות מורשים לעבור. </li><li style=";text-align:right;direction:rtl"> שליח נשלח למטה כמה דקות מותר לעבור על מנת להבטיח כי כל קרוב בקבוקים. </li><li style=";text-align:right;direction:rtl"> הקו הוא התנודד פנימה את הבקבוקים הם פרקו כפי שהם באים לעלות על המדפים. </li><li style=";text-align:right;direction:rtl"> עבור במעמקי הדורשים דגימות 20L עבור DNA ו-RNA, שני בקבוקי Niskin נאספים. אחת הבקבוק משמש המדגם פרמטרים כימיים (ראה להלן), ואילו השני הוא מרוקן לתוך בקבוק. RNA נאסף מיד Niskin השני. יתר הראשון הוא מרוקן גם לתוך בקבוק לתת ~ הנפח הסופי 20L. עבור ברזולוציה נמוכה דגימות 1 ליטר רק מים היא להפריש עבור סינון סטרילי בתוך בקבוקי Nalgene. </li></ol> חלק 2: מדגם אוסף גז <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> אם דגימות חמצן יש צורך, הם נאספים הראשון. </li><li style=";text-align:right;direction:rtl"> לצינור דק המחובר Niskin / Go-פלו ברז ואת הקצה השני של הצינור ממוקם במקום לתוך בקבוקי זכוכית בסרום. </li><li style=";text-align:right;direction:rtl"> בקבוק סרום הוא שטף 3 פעמים עם מים המדגם, מלא עד אפס מקום כזה אין בועות אוויר נוכחים. </li><li style=";text-align:right;direction:rtl"> 50μl כלוריד כספית (HgCl 2) מתווספים הבקבוק בסרום באופן לא להכניס בועות אוויר לתוך המדגם. </li><li style=";text-align:right;direction:rtl"> הבקבוק סגור עם פקק גומי crimped עם מכסה אלומיניום, מאוחסן אז על קרח בצידנית. </li></ol> חלק 3: אוסף מדגם מזין <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מזרק 60 סמ&quot;ק היא שטפה עם מים המדגם, (0.2 מיקרומטר) מחוברת בקו, וכמה מים נדחף דרך acrodisc. </li><li style=";text-align:right;direction:rtl"> בקבוק פוליאתילן היא שטפה עם מים מסוננים 10 ~ מ&quot;ל 3 פעמים (לנער בקבוק כאשר שטיפה). </li><li style=";text-align:right;direction:rtl"> מים מסוננים מדגם מתווסף הבקבוקים, על ¾ מלא. </li><li style=";text-align:right;direction:rtl"> מדגם מאוחסן על קרח במקרר. </li></ol> חלק 4: מדגם אוסף אמוניה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> צינורות לדוגמה הם שטופים במים המדגם. </li><li style=";text-align:right;direction:rtl"> דוגמיות מלאים לחלוטין עם מים המדגם. </li><li style=";text-align:right;direction:rtl"> 5 מ&quot;ל מים מדגם נלקחים החוצה באמצעות pipet 10 מ&quot;ל; את שאר המים בצינור נמחקת לבין 5 מ&quot;ל בקצה pipet מוחלפים לתוך הצינור המדגם. </li><li style=";text-align:right;direction:rtl"> 7.5ml של מגיב אמוניום עובד מתווסף צינור כל פעם דגימות מן המעמקים כל נאספו. </li></ol> חלק 5: אוסף לדוגמה עבור ספירה תא <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> טרום שכותרתו 50 מ&quot;ל צינור פלקון המכיל 5.4 מ&quot;ל 37% פורמלדהיד מלא לראש ריכוז סופי של 4%. הטיפול נלקח לא לשפוך. </li></ol> חלק 6: אוסף דוגמאות למדידות מימן גפרתי <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> טרום שכותרתו 50 מ&quot;ל צינור פלקון המכיל אבץ אצטט 20% (Zn (O 2 CCH 3) 2) מלא למעלה. הטיפול נלקח לא לשפוך. </li></ol> חלק 7: אוסף נפח קטן ריכוז ביומסה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> 1 ליטר ובקבוקים מלאים למעלה. </li></ol> חלק 8: אוסף נפח גדול ריכוז ביומסה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> 20 ליטר carboys מלאים. </li></ol> חלק 9: תוצאות נציג התוצאה הסופית של מי ים הדגימה תוצאות כ -12 דגימות החמצן המומס, 32-48 דגימות גז, 16 דגימות חומר מזין, 32 דגימות אמוניום, 16 דגימות מימן גופרי, 16 דגימות ספירת תאים, 6 20 ליטר carboys, 1 16 בקבוקי ליטר ו 4 Sterivex 0.22μm מסננים המכיל חיידקים ביומסה נשמר עם RNA מאוחר יותר.

<ol>
	<li>Cline, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectrophotometric+determination+of+hydrogen+sulfide+in+natural+waters.">Spectrophotometric determination of hydrogen sulfide in natural waters.</a> Limnology &#38; Oceanography. 14, 454-458 (1969).</li><li>Holmes, R. M., Aminot, A., Kerouel, R., Hooker, B. A., Peterson, B. 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+simple+and+precise+method+for+measuring+ammonium+in+marine+and+freshwater+ecosystems.">A simple and precise method for measuring ammonium in marine and freshwater ecosystems.</a> Canadian Journal of fisheries and Aquatic Science. 56, 1801-1808 (1999).</li></ol>

The authors have nothing to disclose.

בסך הכל, יש הרבה הכנה ותכנון הנדרש עבור סוג זה של הדגימה, כך 2 ימים בתקציב לפני הנסיעה כדי לקבל את הדברים כדי ויום אחד לאחר מכן לניקוי הכל. באופן כללי, לא נותנים מי מלח באים במגע עם המשאבה peristaltic או לתוך תיבת אלומיניום Zarges ששימש להובלת הציוד בתחום. פנקו את הציוד בכבוד זה יהיה ביצועים טובים הרבה זמן לבוא.

<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>20L Carboys</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>02-960-15</td>
<td>&nbsp;</td>
<tr>
<td>3/8" Tygon</td>
<td>&nbsp;</td>
<td>Cole Parmer</td>
<td>06429-36</td>
<td>&nbsp;</td>
<tr>
<td>Sterivex filters</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>SVG010RS</td>
<td>&nbsp;</td>
<tr>
<td>Prefilter Housing Unit</td>
<td>&nbsp;</td>
<td>AMD</td>
<td>501200</td>
<td>&nbsp;</td>
<tr>
<td>Luer Fitting</td>
<td>&nbsp;</td>
<td>Cole Parmer</td>
<td>31507-27</td>
<td>&nbsp;</td>
<tr>
<td>1/4" Tygon tubing</td>
<td>&nbsp;</td>
<td>Cole Parmer</td>
<td>06429-24</td>
<td>&nbsp;</td>
<tr>
<td>1/4" Tygon tubing</td>
<td>&nbsp;</td>
<td>Cole Parmer</td>
<td>06429-24</td>
<td>&nbsp;</td>
<tr>
<td>1/4" Tygon tubing</td>
<td>&nbsp;</td>
<td>Cole Parmer</td>
<td>06429-24</td>
<td>&nbsp;</td>
<tr>
<td>50ml Plastic tubes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Glass Serum Bottles</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Glass Sample Collection</td>
</tr>
<tr>
<td>Septa and aluminum caps for glass serum bottles</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Hand Crimper</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>20-200ul Pipette and Tips</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Glass Tubes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Ammonium Sample Collection</td>
</tr>
<tr>
<td>30ml Plastic Bottles</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Nutrient Collection</td>
</tr>
<tr>
<td>Acrodisc Filters</td>
<td>&nbsp;</td>
<td>Millipore</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>5 ml Tubes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Nitrite Sample Collection</td>
</tr>
<tr>
<td>37% Formaldehyde</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For cell count samples</td>
</tr>
<tr>
<td>20% Zinc Acetate</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For sulfide samples</td>
</tr>
<tr>
<td>Mercuric Chloride</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For gas samples</td>
</tr>
<tr>
<td>milliQ water</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ammonium Working Reagent</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Borate buffer, sodium sulfite, orthophaladialdehyde</td>
</tr>
<tr>
<td>Sulfanilamide</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>5g into 450ml water and 50ml HCl</td>
</tr>
<tr>
<td>N-(1-naphthy)-ethylenediamine dihydrochloride solution</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.5g into 500ml water</td>
</tr>
</tbody>
</table>

seawater sampling and collection

סרט הווידאו הזה מתעד שיטות לאיסוף דגימות מים חוף הים ועיבוד אותם במורד הזרם עבור יישומים שונים, כולל ריכוז ביומסה, טיהור חומצות גרעין, שפע תאים, מזין ומנתח גז עקבות. לקבלת דוגמיות הפגנה של היום נאספו סיפון ההפעלה ג&#39;ון סטריקלנד HMS ב Inlet Saanich. מסגרת A-עגורן, כננת עם רב תכליתי מערכת הכבלים, משמש בשילוב עם Niskin או Go-פלו הדגימה בקבוקי מים. מוליכות, טמפרטורה, ואת עומק (CTD) חיישנים משמשים גם כדי לדגום את מסת המים הבסיסית. כדי למזער outgassing, דגימות גז עקבות נאספים הראשון. לאחר מכן, מזינים, כימיה מים, ספירת תאים נחושים. לבסוף, המים נאספים לסינון ביומסה. בפעם הגדרת אוסף של השחקנים יחיד ~ 1.5 שעות בעומק מרבי של 215 מטרים. לכן, סכום כולל של 6 שעות יש צורך בדרך כלל כדי להשלים את סדרת אוסף המתואר כאן.

Watch this Scientific Journal Video about Seawater Sampling and Collection at JoVE.com

Seawater Sampling and Collection

סרט הווידאו הזה מתעד שיטות לאיסוף דגימות מים חוף הים ועיבוד אותם במורד הזרם עבור יישומים שונים, כולל ריכוז ביומסה, טיהור חומצות גרעין, שפע תאים, מזין ומנתח גז עקבות.

מי ים דגימה אוסף

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass ...

Nanyang Technological University

Research

JoVE Journal

Biology

19.9K Views.  University of British Columbia - UBC. סרט הווידאו הזה מתעד שיטות לאיסוף דגימות מים חוף הים ועיבוד אותם במורד הזרם עבור יישומים שונים, כולל ריכוז ביומסה, טיהור חומצות גרעין, שפע תאים, מזין ומנתח גז עקבות.

Video: Seawater Sampling and Collection

Gibberella zeae Ascospore Production and Collection for Microarray Experiments.

Fusarium graminearum Schwabe (teleomorph Gibberella zeae) is a plant pathogen causing scab disease on wheat and barley that reduces crop yield and grain quality. F. graminearum also causes stalk and ear rots of maize and is a producer of mycotoxins such as the trichothecenes that contaminate grain and are harmful to humans and livestock (Goswami and Kistler, 2004).

The fungus produces two types of spores. Ascospores, the propagules resulting from sexual reproduction, are the main source of primary infection.  These spores are forcibly discharged from mature perithecia and dispersed by wind (Francl et al 1999). Secondary infections are mainly caused by macroconidia which are produced by asexual means on the plant surface. To study the developmental processes of ascospores in this fungus, a procedure for their collection in large quantity under sterile conditions was required. Our protocol was filmed in order to generate the highest level of information for understanding and reproducibility; crucial aspects when full genome gene expression profiles are generated and interpreted. In particular, the variability of ascospore germination and biological activity are dependent on the prior manipulation of the material. The use of video for documenting every step in ascospore production is proposed in order to increase standardization, complying with the increasingly stringent requirements for microarray analysis. The procedure requires only standard laboratory equipment. Steps are shown to prevent contamination and favor time synchronization of ascospores.

To study the developmental processes of ascospores in Gibberella zeae, a procedure for collection under sterile conditions is filmed in order to generate the highest level of information for protocol description. This should facilitate the reproducibility of the experiment, a crucial aspect when full genome expression profile tests are implemented.

Gibberella zeae Ascospore ייצור אוסף לניסויים microarray.

Fusarium graminearum Schwabe (teleomorph Gibberella zeae) is a plant pathogen causing scab disease on wheat and barley that reduces crop yield and grain ...

Collection and Cryopreservation of Hamster Oocytes and Mouse Embryos

Embryos and oocytes were first successfully cryopreserved more than 30 years ago, when Whittingham et al. 1 and Wilmut 2 separately described that mouse embryos could be frozen and stored at -196 &deg;C and, a few years later, Parkening et al. 3 reported the birth of live offspring resulting from in vitro fertilization (IVF) of cryopreserved oocytes. Since then, the use of cryopreservation techniques has rapidly spread to become an essential component in the practice of human and animal assisted reproduction and in the conservation of animal genetic resources. Currently, there are two main methods used to cryopreserve oocytes and embryos: slow freezing and vitrification. A wide variety of approaches have been used to try to improve both techniques and millions of animals and thousands of children have been born from cryopreserved embryos. However, important shortcomings associated to cryopreservation still have to be overcome, since ice-crystal formation, solution effects and osmotic shock seem to cause several cryoinjuries in post-thawed oocytes and embryos. Slow freezing with programmable freezers has the advantage of using low concentrations of cryoprotectants, which are usually associated with chemical toxicity and osmotic shock, but their ability to avoid ice-crystal formation at low concentrations is limited. Slow freezing also induces supercooling effects that must be avoided using manual or automatic seeding 4. In the vitrification process, high concentrations of cryoprotectants inhibit the formation of ice-crystals and lead to the formation of a glasslike vitrified state in which water is solidified, but not expanded. However, due to the toxicity of cyroprotectants at the concentrations used, oocytes/embryos can only be exposed to the cryoprotectant solution for a very short period of time and in a minimum volume solution, before submerging the samples directly in liquid nitrogen 5. In the last decade, vitrification has become more popular because it is a very quick method in which no expensive equipment (programmable freezer) is required. However, slow freezing continues to be the most widely used method for oocyte/embryo cryopreservation. In this video-article we show, step-by-step, how to collect and slowly freeze hamster oocytes with high post-thaw survival rates. The same procedure can also be applied to successfully freeze and thaw mouse embryos at different stages of preimplantation development.

In this video-article we present a step-by-step demonstration on how to collect and cryopreserve hamster oocytes with high post-thaw survival rates. The same procedure can also be applied to successfully freeze and thaw mouse embryos at different stages of preimplantation development.

איסוף cryopreservation של ביציות ועוברים אוגר עכבר

Embryos and oocytes were first successfully cryopreserved more than 30 years ago, when Whittingham et al. 1 and Wilmut 2 separately described that mouse ...

Large Volume (20L+) Filtration of Coastal Seawater Samples

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents large volume (&ge;20 L) filtration of microbial biomass, ranging between 0.22&mu;m and 2.7&mu;m in diameter, from the water column. Two 20L samples can be filtered simultaneously using a single pump unit equipped with four rotating heads. Filtration is done in the field on extended trips, or immediately upon return for day trips. It is important to record the amount of water passing through each sterivex filter unit. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use. This procedure will take approximately 5 hours plus an additional hour for clean up.

Large Volume 20L+ Filtration of Coastal Seawater Samples

This video documents large volume (&ge;20 L) filtration of microbial biomass, ranging between 0.22&mu;m and 2.7&mu;m in diameter, from the water column.

נפח גדול (20L +) סינון של דגימות מי ים החוף

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, ...

Small Volume (1-3L) Filtration of Coastal Seawater Samples

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents small volume (~1 L) filtration of microbial biomass from the water column. The protocol is an extension of the large volume sampling protocol described earlier, with one major difference: here, there is no pre-filtration step, so all size classes of biomass are collected down to the 0.22 &mu;m filter cut-off. Samples collected this way are ideal for nucleic acid analysis. The set-up, filtration, and clean-up steps each take about 20-30 minutes. If using two peristaltic pumps simultaneously, up to 8 samples may be filtered at the same time. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use.

Small Volume 1-3L Filtration of Coastal Seawater Samples

This video documents small volume (~1 L) filtration of microbial biomass from the water column.

נפח קטן (1-3L) סינון של דגימות מי ים החוף

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s ...

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

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 colony. Proper collection of tissues for further histological processing can impact the quality of testing results. This article describes the conduct of a basic gross examination including identification of heart, liver, lungs, kidneys, and spleen, as well as how to collect those organs. Additionally four of the more difficult tissue/sample collection techniques are demonstrated. Lung collection and perfusion can be particularly challenging as the tissue needs to be properly inflated with a fixative in order for inside of the tissue to fix properly and to enable thorough histologic evaluation. This article demonstrates the step by step technique to remove the lung and inflate it with fixative in order to achieve optimal fixation of the tissue within 24 hours. Brain collection can be similarly challenging as the tissue is soft and easily damaged. This article demonstrates the step by step technique to expose and remove the brain from the skull with minimal damage to the tissue. The mesenteric lymph node is a good sample type in which to detect many common infectious agents as enteric viruses persist longer in the lymph node than they are shed in feces. This article demonstrates the step by step procedure for locating and aseptically removing the mesenteric lymph node. Finally, identification of infectious agents of the respiratory tract may be performed by bacterial culture or PCR testing of nasal and/or bronchial fluid aspirates taken at necropsy. This procedure describes obtaining and preparing the respiratory aspirate sample for bacterial culture and PCR testing.

This article describes the procedures for conducting a basic postmortem examination of a mouse or rat, and the collection of basic organs, as well as more challenging sample types from for histological, microbiological, and PCR evaluation.

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 ...

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 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 ...

Collection and Analysis of Arabidopsis Phloem Exudates Using the EDTA-facilitated Method

The plant phloem is essential for the long-distance transport of (photo-) assimilates as well as of signals conveying biotic or abiotic stress. It contains sugars, amino acids, proteins, RNA, lipids and other metabolites. While there is a large interest in understanding the composition and function of the phloem, the role of many of these molecules and thus, their importance in plant development and stress response has yet to be determined. One barrier to phloem analysis lies in the fact that the phloem seals itself upon wounding. As a result, the number of plants from which phloem sap can be obtained is limited. One method that allows collection of phloem exudates from several plant species without added equipment is the EDTA-facilitated phloem exudate collection described here. While it is easy to use, it does lead to the wounding of cells and care has to be taken to remove contents of damaged cells. In addition, several controls to prove purity of the exudate are necessary. Because it is an exudation rather than a direct collection of the phloem sap (not possible in many species) only relative quantification of its contents can occur. The advantage of this method over others is that it can be used in many herbaceous or woody plant species (Perilla, Arabidopsis, poplar, etc.) and requires minimal equipment and training. It leads to reasonably large amounts of exudates that can be used for subsequent analysis of proteins, sugars, lipids, RNA, viruses and metabolites. It is simple enough that it can be used in both a research as well as in a teaching laboratory.

Collection and Analysis of Arabidopsis Phloem Exudates Using the EDTA-facilitated Method

Knowledge of the composition of the phloem sap as well as the mechanism of its loading and long-distance transport is essential for the understanding of long-distance signaling in plant development and stress/pathogen response and of assimilate transport. This manuscript describes the collection of phloem exudates using the EDTA-facilitated method.

איסוף וניתוח של<em&gt; ארבידופסיס</em&gt; השיפה exudates שימוש בשיטת EDTA-הקלה

The plant phloem is essential for the long-distance transport of (photo-) assimilates as well as of signals conveying biotic or abiotic stress. It ...

Non-Terminal Blood Sampling Techniques in Guinea Pigs

Guinea pigs possess several biological similarities to humans and are validated experimental animal models1-3. However, the use of guinea pigs currently represents a relatively narrow area of research and descriptive data on specific methodology is correspondingly scarce. The anatomical features of guinea pigs are slightly different from other rodent models, hence modulation of sampling techniques to accommodate for species-specific differences, e.g., compared to mice and rats, are necessary to obtain sufficient and high quality samples. As both long and short term in vivo studies often require repeated blood sampling the choice of technique should be well considered in order to reduce stress and discomfort in the animals but also to ensure survival as well as compliance with requirements of sample size and accessibility. Venous blood samples can be obtained at a number of sites in guinea pigs e.g., the saphenous and jugular veins, each technique containing both advantages and disadvantages4,5. Here, we present four different blood sampling techniques for either conscious or anaesthetized guinea pigs. The procedures are all non-terminal procedures provided that sample volumes and number of samples do not exceed guidelines for blood collection in laboratory animals6. All the described methods have been thoroughly tested and applied for repeated in vivo blood sampling in studies within our research facility.

Though a known model, the guinea pig currently represents a niche in experimental animal sciences and limited data is available on the execution of most procedures. Here we present four different approaches to non-terminal in vivo blood sampling techniques in either conscious or anaesthetized guinea pigs.

טכניקות דגימת דם ללא מסוף בגינאה חזירים

Guinea pigs possess several biological similarities to humans and are validated experimental animal models1-3. However, the use of guinea pigs currently ...

Repeated Blood Collection for Blood Tests in Adult Zebrafish

Repeated blood collection is one of the most common techniques performed on laboratory animals. However, a non-lethal protocol for blood collection from zebrafish has not been established. The previous methods for blood collection from zebrafish are lethal, such as lateral incision, decapitation and tail ablation. Thus we have developed a novel &#8220;repeated&#8221; blood collection method, and present here a detailed protocol outlining this procedure. This method is minimally invasive and results in a very low mortality rate (2.3%) for zebrafish, thus enabling repeated blood sampling from the same individual. The maximum volume of blood sampling is dependent on body weight of the fish. The volume for repeated blood sampling at intervals should be&#160;&#8804;0.4% of body weight every week or &#8804;1% every 2 weeks, which were evaluated by measurements of blood hemoglobin. Additionally, hemoglobin, fasting blood glucose, plasma triacylglycerol (TG) and total cholesterol levels in male and female adult zebrafish were measured. We also applied this method to investigate the dysregulation of glucose metabolism in diet-induced obesity. This blood collection method will allow many applications, including glucose and lipid metabolism and hematological studies, which will increase the use of zebrafish as a human disease model organism.

Repeated blood sampling is necessary and important in animal research. We developed a novel, non-lethal and reliable method for repeated blood collection from adult zebrafish, and applied this method to the study of blood biochemistry, including glucose metabolism.

איסוף דם חוזרים ונשנים לבדיקות דם בדג הזברה למבוגרים

Repeated blood collection is one of the most common techniques performed on laboratory animals. However, a non-lethal protocol for blood collection from ...

Gamete Collection and In Vitro Fertilization of Astyanax mexicanus

Astyanax mexicanus is emerging as a model organism for a variety of research fields in biological science. Part of the recent success of this teleost fish species is that it possesses interfertile cave and river-dwelling populations. This enables the genetic mapping of heritable traits that were fixed during adaptation to the different environments of these populations. While this species can be maintained and bred in the lab, it is challenging to both obtain embryos during the daytime and create hybrid embryos between strains. In vitro fertilization (IVF) has been used with a variety of different model organisms to successfully and repeatedly breed animals in the lab. In this protocol, we show how, by acclimatizing A. mexicanus to different light cycles coupled with changes in water temperature, we can shift breeding cycles to a chosen time of the day. Subsequently, we show how to identify suitable parental fish, collect healthy gametes from males and females, and produce viable offspring using IVF. This enables related procedures such as the injection of genetic constructs or developmental analysis to occur during normal working hours. Furthermore, this technique can be used to create hybrids between the cave and surface-dwelling populations, and thereby enable the study of the genetic basis of phenotypic adaptations to different environments.

Gamete Collection and In Vitro Fertilization of Astyanax mexicanus

In vitro fertilization is a commonly used technique with a variety of model organisms to maintain lab populations and produce synchronized embryos for downstream applications. Here, we present a protocol that implements this technique for different populations of the Mexican tetra fish, Astyanax mexicanus.

אוסף המשחק ו הפריה חוץ גופית של Astyanax מקסיאנוס

Astyanax mexicanus is emerging as a model organism for a variety of research fields in biological science. Part of the recent success of this teleost fish ...