الشكر الخاص لساندرز كوري في تصاميم يروك لإعداد الرسوم التوضيحية وكريس Reddi في جامعة كاليفورنيا لإقامة الثقافات عينة للشخصيات. تم توفير التمويل لهذا المشروع من قبل HHMI (HHMI منحة رقم 52006944).

<ol>
	<li>Barker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> At the Bench: A Laboratory Navigator. , (1998).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biosafety in Microbiological And Biomedical Laboratories (BMBL). , (2009).</li><li>Bykowski, T., Stevenson, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aseptic+Technique.">Aseptic Technique.</a> Current Protocols in Microbiology. Appendix 4, Appendix 4D (2008).</li><li>Cot&#233;, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aseptic+Technique+for+Cell+Culture.">Aseptic Technique for Cell Culture.</a> Current Protocols in Cell Biology. Chapter 1, Unit 1.3 (2001).</li><li>Grimes, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+basic+laboratory+manual+for+the+small-scale+production+and+testing+of+I-2+Newcastle+disease+vaccine.">A basic laboratory manual for the small-scale production and testing of I-2 Newcastle disease vaccine.</a> RAP Publication. AC802/E, 12-13 (2002).</li><li>Guzman, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pipetting:+A+Practical+Guide.">Pipetting: A Practical Guide.</a> The American Biology Teacher. 63 (2), 128-131 (2001).</li><li>Jordan, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> RESEARCH: National Genomics Research Initiative Phage Resource Laboratory Manual. , (2008).</li><li>Sambrook, J., Russell, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Molecular Cloning - A Laboratory Manual. , 978-087969576 (2001).</li><li>Seidman, L. A., Moore, C. 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> Basic Laboratory Methods for Biotechnology: Textbook and Laboratory Reference. , (2000).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> PIPETMAN P User's Guide. , (2012).</li></ol>

ليس لدي ما يكشف.

تقنية العقيم ويشير إلى مجموعة من الإجراءات الروتينية القيام به لمنع الحلول العقيمة والثقافات من أن تصبح ملوثة غير مرغوب فيه من قبل الكائنات الحية الدقيقة في المختبر. هذه التقنيات ضرورية لإجراء التجارب التي تتطلب الخلايا المتنامية. على الرغم من عدم إعداد العمل الذي هو عقيم تماما أن يتحقق، والإجراءات مثل تعقيم السطوح مختبر، وخلق مجال معقم باستخدام موقد بنسن، والحد من التعرض للثقافات لم يسبق لهم اللعب ووسائل الإعلام في الهواء، وتعقيم المواد مثل الأنابيب والزجاجات والماصات الزجاجية، وتجنب الاتصال مع الأدوات المعقمة غير معقمة أسطح يقلل من احتمال تلوث الحلول والثقافات في تجربة. والهدف من ذلك هو لهذه الإجراءات الاحترازية لتصبح طبيعة ثانية، وهذا يأتي مع التدريب والممارسة بينما كان يعمل في المختبر. نقل وحدة التخزين مع حلول عقيمة والثقافات باستخدام أدوات مثل الماصات المصلية وهيئة التصنيع العسكريropipettors هي واحدة من العديد من أنواع تقنيات روتين عمله في أحد المختبرات. التطبيقات التجريبية المختلفة استدعاء لأدوات قادر على نقل متميزة، وأحجام دقيقة بعد دقيقة و،. وتستخدم ماصات المصلية في مختبرات علم الأحياء الدقيقة لإعداد مزارع الخلايا التي تتطلب استعدادات وسائل الإعلام التي تشمل وحدات التخزين ملليلتر، بينما micropipettors ضرورية لتجارب البيولوجيا الجزيئية التي تحتاج إلى كميات ميكروليتر فقط من الحلول. عندما يمارس أسلوب العقيم مع هذه الصكوك، والتقليل من التلوث أثناء عمليات النقل بغض النظر عن حجم كمية من السائل أو النوع من التجارب. على الرغم من عدم مناقشتها في هذا البروتوكول، واحد وسيلة أخرى تستخدم عادة لمنع التلوث هو العمل داخل غطاء محرك السيارة تدفق رقائقي (الشكل 10). هذه المعدات هو أمر حاسم لزراعة الأنسجة وللتجارب أجريت مع الكائنات الحية الدقيقة المصنفة BLS-2 أو أعلى. غطاء تدفق رقائقي يحتوي على (HEPA ذات الكفاءة العاليةالهواء جسيمات) مرشح أن يزيل الملوثات المحمولة جوا من الجو التي تصب في غطاء محرك السيارة في حين منع الهواء فلتر من غرفة من تتخلل مساحة العمل. من المذكرة، لا يمكن أن تستخدم موقد بنسن داخل الصفحي غطاء محرك السيارة، لأن تدفق الحرارة من لهب يعطل تدفق الهواء أساسي إلى وظيفة غطاء محرك السيارة. غالبا ما يكون من المفيد للتحقق من جودة الأسلوب الخاص بك العقيم عند تنفيذ التجارب العملية. للتأكد من الحلول وليس وسائل الإعلام ثقافة للتلوث خلال التلاعب التجريبية، وإعداد عنصر تحكم دائما سلبية. على سبيل المثال، إذا كان إعداد أنابيب من مرق لنمو الثقافات البكتيرية، لا تطعيم أنبوب واحد ترك وسائل الاعلام فقط العقيمة. احتضان والمتوسطة إلى جانب أنابيب تلقيح ثم تفقد السيطرة أنبوب uninoculated بحثا عن علامات على تلوث مثل تعكر من نمو الخلايا غير المرغوب فيها وعرض عن غير قصد في أنبوب. إذا كانت ملوثة أنبوب السيطرة، وأنبوب التجريبيةمن المرجح تكون ملوثة أيضا، وهذه التجربة لا بد من تكرارها. وينبغي القيام بهذه التدابير الاحترازية مع كل تجربة.

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> LB حساء </td><td> Difco </td><td> 244620 </td><td> تتوفر أيضا وصفة في إشارة 6 </td></tr><tr><td colspan="4"> TE العازلة: </td></tr><tr><td> EDTA ثنائي الصوديوم ملح ثنائي الهيدرات </td><td> سيغما </td><td> E5134 </td><td></td></tr><tr><td> Trizma-حمض الهيدروكلوريك </td><td> سيغما </td><td> T-3253 </td><td></td></tr><tr><td> CiDecon </td><td> التطهير المختبرات، وشركة </td><td> 8504 </td><td> مطهر </td></tr><tr><td> الإيثانول </td><td> فيشر العلمية </td><td> A406 </td><td> لاستخدامها كعامل مطهر، وإعداد 70٪ (V / V) مع الماء المقطر </td></tr></tbody></table>

aseptic laboratory techniques: volume transfers with serological pipettes and micropipettors

الكائنات الحية الدقيقة في كل مكان - في الهواء والتربة، وجسم الإنسان وكذلك على الأسطح غير حية مثل المقاعد ومختبر لوحة مفاتيح الحاسوب. في كل مكان من الميكروبات يخلق امدادات وفيرة من الملوثات المحتملة في المختبر. لضمان نجاح التجربة، يجب أن يكون الحد الأدنى لعدد من الملوثات على الأسطح والمعدات والعمل. المشترك بين العديد من التجارب في علم الأحياء المجهرية من التقنيات التي تنطوي على قياس ونقل الثقافات التي تحتوي على الخلايا البكتيرية أو الجسيمات الفيروسية. للقيام بذلك من دون الاتصال غير معقمة أو الأسطح الملوثة وسائل الاعلام عقيمة يتطلب (1) إعداد مساحة عمل عقيم، (2) تحدد بدقة وقراءة بدقة الأدوات اللازمة لنقل العقيم من السوائل، و (3) معالجة بشكل صحيح من الصكوك والثقافات قوارير وزجاجات و أنابيب داخل حقل معقمة. تعلم هذه الإجراءات تدعو إلى تدريب وممارسة. في البداية، وينبغي اتخاذ إجراءات قد تكون بطيئة، مدروسة، والتي تسيطر عليها مع هدف يجري لتي العقيمechnique لتصبح طبيعة ثانية عند العمل على مقاعد البدلاء. هنا نقدم خطوات لقياس حجم استخدام ماصات المصلية وmicropipettors داخل حقل العقيمة التي أنشأتها ناسخ بنسن. أحجام تتراوح ما بين microliters (ميكرولتر) إلى ملليلتر (مل) اعتمادا على الأداة المستخدمة. نقل السوائل عادة تشمل حساء عقيم أو المحاليل الكيميائية، وكذلك الثقافات البكتيرية وأسهم فج. باتباع هذه الإجراءات، يجب أن يكون الطلاب قادرين على: <ul style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> العمل في مجال العقيمة التي أنشأتها لهب الموقد بنسن. </li><li style=";text-align:right;direction:rtl"> استخدام ماصات المصلية دون المساس عقم الصك. </li><li style=";text-align:right;direction:rtl"> نضح السوائل مع ماصات المصلية، والقراءة على وجه التحديد كميات معايرة عن طريق محاذاة الغضروف المفصلي التي شكلها السائل إلى علامات التخرج على ماصة. </li><li style=";text-align:right;direction:rtl"> الحفاظ على الزجاجات والثقافة، وقوارير وأنابيب وقبعات كل منهما معقم خلال نقل السائل. </li><li style=";text-align:right;direction:rtl"> التعرف على التطبيقات المختلفة لبيب المصلية البلاستيك مقابل زجاجettes. </li><li style=";text-align:right;direction:rtl"> قيود دقة الدولة للmicropipettors. </li><li style=";text-align:right;direction:rtl"> على وجه التحديد وبدقة وضع وحدات التخزين على micropipettors. </li><li style=";text-align:right;direction:rtl"> معرفة كيفية استخدام صحيح المحطة الأولى والثانية على micropipettor إلى نضح ونقل وحدات التخزين الصحيح. </li></ul>

Watch this Scientific Journal Video about Aseptic Laboratory Techniques: Volume Transfers with Serological Pipettes and Micropipettors at JoVE.com

Aseptic Laboratory Techniques: Volume Transfers with Serological Pipettes and Micropipettors

عند العمل في المختبر، لا بد من الحد من مصادر التلوث. تقنية العقيم يشير إلى الإجراءات التي تسمح لنقل الثقافات والكواشف مع تجنب الاتصال مع المنظمات غير معقمة السطوح. وتستخدم ماصات المصلية وmicropipettors لقياس كميات الدقيق دون المساس عقم الحلول المستخدمة في التجارب.

مختبر تقنيات العقيم: التحويلات الحجم مع الماصات المصلية وMicropipettors

Microorganisms are everywhere - in the air, soil, and human body as well as on inanimate surfaces like laboratory benches and computer keyboards. The ...

aseptic-laboratory-techniques-volume-transfers-with-serological

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Biology

161.8K Views.  University of California, Los Angeles. عند العمل في المختبر، لا بد من الحد من مصادر التلوث. تقنية العقيم يشير إلى الإجراءات التي تسمح لنقل الثقافات والكواشف مع تجنب الاتصال مع المنظمات غير معقمة السطوح. وتستخدم ماصات المصلية وmicropipettors لقياس كميات الدقيق دون المساس عقم الحلول المستخدمة في التجارب.

Video: Aseptic Laboratory Techniques: Volume Transfers with Serological Pipettes and Micropipettors

Microbial Communities in Nature and Laboratory - Interview

المجتمعات الميكروبية في الطبيعة ومختبر -- مقابلة

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades. Today, such pipettes are made from glass capillaries using a programmable puller. Such instruments heat the capillary using either a metal filament or a laser and draw out the glass using gravity, a motor or both. Pipettes for patch-clamp recording are formed using only heat and gravity, while sharp electrodes for intracellular recording use a combination of heat, gravity, and a motor. The procedure used to make intracellular recording pipettes is similar to that used to make injection needles for a variety of applications, including cRNA injection into Xenopus oocytes. In general, capillary glass &lt;1.2 mm in diameter is used to make pipettes for patch clamp recording, while narrower glass is used for intracellular recording (outer diameter = 1.0 mm). For each tool, the puller is programmed slightly differently. This video shows how to make both kinds of recording pipettes using pre-established puller programs.

This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.

جعل التصحيح ، وماصات أقطاب شارب مع بولير برمجة

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

Pressure-polishing Pipettes for Improved Patch-clamp Recording

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording. We first developed this method for fabricating pipettes suitable for recording from small (&lt;3 m) neuronal cell bodies. The basic principal is similar to glass-blowing and combines air pressure and heat to modify the shape of patch pipettes prepared by a conventional micropipette puller. It can be applied to so-called soft (soda lime) and hard (borosilicate) glasses. Generally speaking, pressure polishing can reduce pipette resistance by 25% without decreasing the diameter of the tip opening (Goodman and Lockery, 2000). It can be applied to virtually any type of glass and requires only the addition of a high-pressure valve and fitting to a microforge. This technique is essential for recording from ultrasmall cells (&lt;5 m) and can also improve single-channel recording by minimizing pipette resistance. The blunt shape is also useful for perforated-patch clamp recording since this tip shape results in a larger membrane bleb available for perforation.

This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.

الضغط تلميع الماصات لتسجيل باتش تحسين المشبك

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Modeling Biological Membranes with Circuit Boards and Measuring Electrical Signals in Axons: Student Laboratory Exercises

This is a demonstration of how electrical models can be used to characterize biological membranes. This exercise also introduces biophysical terminology used in electrophysiology. The same equipment is used in the membrane model as on live preparations. Some properties of an isolated nerve cord are investigated: nerve action potentials, recruitment of neurons, and responsiveness of the nerve cord to environmental factors.

This is a demonstration of how biological membranes can be understood using electrical models. We also demonstrate procedures for recording action potentials from the ventral nerve cord of the crayfish for student orientated laboratories.

النمذجة الأغشية البيولوجية مع لوحات الدوائر الكهربائية وقياس الاشارات في محاور عصبية : تمارين مختبر طالبة

This is a demonstration of how electrical models can be used to characterize biological membranes. This exercise also introduces biophysical terminology ...

Aseptic Laboratory Techniques: Plating Methods

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: 
 
&#9679; Perform plating procedures without contaminating media. 
&#9679; Isolate single bacterial colonies by the streak-plating method. 
&#9679; Use pour-plating and spread-plating methods to determine the concentration of bacteria. 
&#9679; Perform soft agar overlays when working with phage. 
&#9679; Transfer bacterial cells from one plate to another using the replica-plating procedure. 
&#9679; Given an experimental task, select the appropriate plating method.

When working with media and reagents used to culture microorganisms, aseptic technique must be practiced to ensure contamination is minimized. A variety of plating methods are routinely used to isolate, propagate, or enumerate bacteria and phage, all of which incorporate procedures that maintain the sterility of experimental materials.

مختبر تقنيات العقيم: طرق تصفيح

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies ...

Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands

Histology volume reconstruction facilitates the study of 3D shape and volume change of an organ at the level of macrostructures made up of cells. It can also be used to investigate and validate novel techniques and algorithms in volumetric medical imaging and therapies. Creating 3D high-resolution atlases of different organs1,2,3 is another application of histology volume reconstruction. This provides a resource for investigating tissue structures and the spatial relationship between various cellular features. We present an image registration approach for histology volume reconstruction, which uses a set of optical blockface images. The reconstructed histology volume represents a reliable shape of the processed specimen with no propagated post-processing registration error. The Hematoxylin and Eosin (H&#38;E) stained sections of two mouse mammary glands were registered to their corresponding blockface images using boundary points extracted from the edges of the specimen in histology and blockface images. The accuracy of the registration was visually evaluated. The alignment of the macrostructures of the mammary glands was also visually assessed at high resolution.
This study delineates the different steps of this image registration pipeline, ranging from excision of the mammary gland through to 3D histology volume reconstruction. While 2D histology images reveal the structural differences between pairs of sections, 3D histology volume provides the ability to visualize the differences in shape and volume of the mammary glands.

We present an image registration approach for 3-dimensional (3D) histology volume reconstruction, which facilitates the study of the changes of an organ at the level of macrostructures made up of cells . Using this approach, we studied the 3D changes between wild-type and Igfbp7-null mammary glands.

إعادة بناء 3 الأبعاد حجم الأنسجة وتطبيقاتها لدراسة ماوس الغدد الثديية

Histology volume reconstruction facilitates the study of 3D shape and volume change of an organ at the level of macrostructures made up of cells. It can ...

Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry

Extracellular Vesicles (EVs) are small, membrane-derived vesicles found in bodily fluids that are highly involved in cell-cell communication and help regulate a diverse range of biological processes. Analysis of EVs using flow cytometry (FCM) has been notoriously difficult due to their small size and lack of discrete populations positive for markers of interest. Methods for EV analysis, while considerably improved over the last decade, are still a work in progress. Unfortunately, there is no one-size-fits-all protocol, and several aspects must be considered when determining the most appropriate method to use. Presented here are several different techniques for processing EVs and two protocols for analyzing EVs using either individual detection or a bead-based approach. The methods described here will assist with eliminating the antibody aggregates commonly found in commercial preparations, increasing signal&#8211;to-noise ratio, and setting gates in a rational fashion that minimizes detection of background fluorescence. The first protocol uses an individual detection method that is especially well suited for analyzing a high volume of clinical samples, while the second protocol uses a bead-based approach to capture and detect smaller EVs and exosomes.

Many different methods exist for the measurement of extracellular vesicles (EVs) using flow cytometry (FCM). Several aspects should be considered when determining the most appropriate method to use. Two protocols for measuring EVs are presented, using either individual detection or a bead-based approach.

تقنيات لتحليل خارج الخلية الحويصلات عن طريق التدفق الخلوي

Extracellular Vesicles (EVs) are small, membrane-derived vesicles found in bodily fluids that are highly involved in cell-cell communication and help ...

Measuring Pressure Volume Loops in the Mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the mouse provides great potential to explore cardiac function at the molecular level. The mouse&#39;s small size does present some challenges in regards to performing detailed cardiac phenotyping. Miniaturization and other advancements in technology have made many methods of cardiac assessment possible in the mouse. Of these, the simultaneous collection of pressure and volume data provides a detailed picture of cardiac function that is not available through any other modality. Here a detailed procedure for the collection of pressure-volume loop data is described. Included is a discussion of the principles underlying the measurements and the potential sources of error. Anesthetic management and surgical approaches are discussed in great detail as they are both critical to obtaining high quality hemodynamic measurements. The principles of hemodynamic protocol development and relevant aspects of data analysis are also addressed.

This manuscript describes a detailed protocol for the collection of pressure-volume data from the mouse.

قياس حجم حلقات الضغط في الماوس

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...

Techniques to Induce and Quantify Cellular Senescence

In response to cellular stress or damage, proliferating cells can induce a specific program that initiates a state of long-term cell-cycle arrest, termed cellular senescence. Accumulation of senescent cells occurs with organismal aging and through continual culturing in vitro. Senescent cells influence many biological processes, including embryonic development, tissue repair and regeneration, tumor suppression, and aging. Hallmarks of senescent cells include, but are not limited to, increased senescence-associated &#946;-galactosidase activity (SA-&#946;-gal); p16INK4A, p53, and p21 levels; higher levels of DNA damage, including &#947;-H2AX; the formation of Senescence-associated Heterochromatin Foci (SAHF); and the acquisition of a Senescence-associated Secretory Phenotype (SASP), a phenomenon characterized by the secretion of a number of pro-inflammatory cytokines and signaling molecules. Here, we describe protocols for both replicative and DNA damage-induced senescence in cultured cells. In addition, we highlight techniques to monitor the senescent phenotype using several senescence-associated markers, including SA-&#946;-gal, &#947;-H2AX and SAHF staining, and to quantify protein and mRNA levels of cell cycle regulators and SASP factors. These methods can be applied to the assessment of senescence in various models and tissues.

Cellular senescence, the irreversible state of cell-cycle arrest, can be induced by various cellular stresses. Here, we describe protocols to induce cellular senescence and methods to assess markers of senescence.

تقنيات للحث وتحديد الخلوي الشيخوخة

In response to cellular stress or damage, proliferating cells can induce a specific program that initiates a state of long-term cell-cycle arrest, termed ...

Volume Segmentation and Analysis of Biological Materials Using SuRVoS (Super-region Volume Segmentation) Workbench

Segmentation is the process of isolating specific regions or objects within an imaged volume, so that further study can be undertaken on these areas of interest. When considering the analysis of complex biological systems, the segmentation of three-dimensional image data is a time consuming and labor intensive step. With the increased availability of many imaging modalities and with automated data collection schemes, this poses an increased challenge for the modern experimental biologist to move from data to knowledge. This publication describes the use of SuRVoS Workbench, a program designed to address these issues by providing methods to semi-automatically segment complex biological volumetric data. Three datasets of differing magnification and imaging modalities are presented here, each highlighting different strategies of segmenting with SuRVoS. Phase contrast X-ray tomography (microCT) of the fruiting body of a plant is used to demonstrate segmentation using model training, cryo electron tomography (cryoET) of human platelets is used to demonstrate segmentation using super- and megavoxels, and cryo soft X-ray tomography (cryoSXT) of a mammalian cell line is used to demonstrate the label splitting tools. Strategies and parameters for each datatype are also presented. By blending a selection of semi-automatic processes into a single interactive tool, SuRVoS provides several benefits. Overall time to segment volumetric data is reduced by a factor of five when compared to manual segmentation, a mainstay in many image processing fields. This is a significant savings when full manual segmentation can take weeks of effort. Additionally, subjectivity is addressed through the use of computationally identified boundaries, and splitting complex collections of objects by their calculated properties rather than on a case-by-case basis.

Volume Segmentation and Analysis of Biological Materials Using SuRVoS Super-region Volume Segmentation Workbench

Segmentation of three-dimensional data from many imaging techniques is a major bottleneck in analysis of complex biological systems. Here, we describe the use of SuRVoS Workbench to semi-automatically segment volumetric data at various length-scales using example datasets from cryo-electron tomography, cryo soft X-ray tomography, and phase contrast X-ray tomography techniques.

حجم تجزئة وتحليل المواد البيولوجية باستخدام منضدة سورفوس (منطقة عظمى تجزئة وحدة التخزين)

Segmentation is the process of isolating specific regions or objects within an imaged volume, so that further study can be undertaken on these areas of ...