From thebrog from hotmail.com Wed Oct 1 11:35:05 2008 From: thebrog from hotmail.com (thebrog@hotmail.com) Date: Wed Oct 1 12:44:52 2008 Subject: [Zbrafish] Re: tank cleaning References: Message-ID: <001862de-b1f9-4729-9ad4-78ede4d261a0@m74g2000hsh.googlegroups.com> On Aug 29, 9:11?pm, Padnos.B...@epamail.epa.gov wrote: > Does anyone have a good method for cleaning polycarbonate tanks? Running > the tanks through our cage washer does not get much, if any, of the > debris off the surfaces. we bleach them in a tank with no less than 10% bleach. then rinse them once let them drip for an hour of so or over night, then rinse them again. job done. can do 35 at a time. andy From gottalovelife from gmail.com Mon Oct 6 13:23:29 2008 From: gottalovelife from gmail.com (Kim) Date: Mon Oct 6 13:25:00 2008 Subject: [Zbrafish] Preparation of enzymes for oxidative stress assays Message-ID: <3cff7cca-923e-4c6d-8d95-baeff28748c1@r15g2000prh.googlegroups.com> I'm trying to run multiple oxidative stress assays for enzymes such as catalase, peroxidase, etc. I am not very familiar with such procedures and was wondering if anyone could provide me with a detailed protocol to prepare the enzymes from embryos to produce quality enzyme extract. Thank you in advance From wendyf from aquaneering.com Wed Oct 8 13:06:01 2008 From: wendyf from aquaneering.com (wendyf@aquaneering.com) Date: Wed Oct 8 13:09:18 2008 Subject: [Zbrafish] Zebrafish Photomicrography Contest Message-ID: <15cbbc15-ebdb-45bb-ac33-5ee831653c15@z18g2000prn.googlegroups.com> 2009 Calendar Contest! For Zebrafish and Xenopus Researchers After the huge success of Aquaneering's 2008 Calendar featuring an image by Sean Megason and Scott Fraser of the California Institute of Technology, we have decided to get everyone involved by sponsoring a Photomicrography contest for Aquatics Researchers for the 2009 calendar year. Your Image in Print on our 2009 Calendar! DEADLINE - NOVEMBER 1, 2008 FANTASTIC PRIZES! Grand Prize: MacBook Laptop Computer 2nd Prize $750 3rd Prize $500 Rules Calendar Contest Rules For 2009 Photomicrography of Zebrafish and Xenopus 1. DEADLINE FOR SUBMISSION is November 1, 2008. Files may be submitted by email (Subject line: Calendar Contest) to:wendyf@aquaneering.com or by mailing a CD to the following address: Aquaneering, Inc., 7960 Stromesa Court, San Diego, CA 92126, ATTN: Calendar Contest. No images will be accepted after midnight on November 10, 2008. 2. All photomicrography images submitted must be created by the entrant depicting studies with zebrafish, xenopus frogs, or other aquatic species raised in a laboratory setting. Up to 2 images may be submitted per person. 3. The following guidelines for file format MUST be followed: Files must be jpeg only, 300ppi, sized 8 x 10 or 8 x 8. Zipped files are acceptable. All images must be accompanied by an entry form, which can be emailed to you. Files must be labeled as follows: LastnameFirstname01.jpg. (or LastnameFirstname02.jpg if 2 files are sent). 4. Description of image(s) must be included on entry form including what disease or condition is being studied, what body part is depicted in the image, and what (if any) dyes were used for labeling. Descriptions may be used for publication. 5. Contest is open to researchers, students (undergrad or graduate) and post docs. No employees of imaging equipment manufacturers or dealers are eligible. No employees of Aquaneering, Inc. or any other manufacturer or dealer of Aquatics Housing are eligible. All entrants must be affiliated with a university, college, medical school, hospital or research facility. 6. Judging will be done by a panel at Aquaneering, Inc. Decision of the judges is final. Award recipients will be notified by December, 2008. 7. PRIZES: First Prize - Laptop Computer (Apple Macbook or PC Laptop) Second Prize - $750 Third Prize - $500 Entry Form Entry Form: please print clearly or cut and paste to a word doc. fill out and return as an attachment to an email or by mail Name________________________________________________________________ Facility_______________________________________________________________ Address______________________________________________________________ _____________________________________________________________________ City____________________________State_____________Zip_________________ Phone (w)________________________Phone (c)____________________________ Email________________________________________________________________ IMAGE 01 DESCRIPTION: (including what disease or condition is being studied, what body part is depicted in the image, and what (if any) dyes were used for labeling - descriptions may be used for publication.) ________________________________________________________________________________ ________________________________________________________________________________ ________________________________________________________________________________ IMAGE 02 DESCRIPTION: ________________________________________________________________________________ ________________________________________________________________________________ ________________________________________________________________________________ I certify that I have produced the image submitted without the help of an instructor. I understand that the file(s) submitted will not be returned to me, and I release all rights for publication of the image(s) to Aquaneering, Inc. Signed__________________________________________Date____________________________ This event is sure to bring out many phenomenal images, so let those creative juices flow! Don't forget the Deadline is November 1, 2008. For more information or questions, please contact: Wendy Porter-Francis Aquaneering, Inc wendyf@aquaneering.com From zfinadmin from gmail.com Wed Oct 8 13:16:42 2008 From: zfinadmin from gmail.com (zfinadmin@gmail.com) Date: Wed Oct 8 13:17:52 2008 Subject: [Zbrafish] Groupleader at the MPI Molecular Cell Biology and Genetics, Dresden Message-ID: <46007654-3ea2-43a7-b11d-f55f098a7516@q26g2000prq.googlegroups.com> Max Planck Institute of Molecular Cell Biology and Genetics The Max Planck Institute of Molecular Cell Biology and Genetics Dresden, Germany is seeking outstanding candidates for Research Group Leader Research at the MPI-CBG focuses on the molecular mechanisms underlying the structure and organization of cells and tissues (see http://www.mpi-cbg.de). In this search we are especially looking for applicants interested in the control of size, shape and/or number at the organelle, cell or tissue level, though all excellent candidates will be considered. We particularly encourage applicants taking biochemical or electron microscopic approaches to cell polarity, the cortex or morphogenesis in model organisms, including the mouse. The working language at the MPI-CBG is English and fluency in English is required. The position is initially for 5 years with the possibility of extension for up to an additional 4 years. The position will be compensated according to the TV?D scale EG15. Funds are available for a postdoctoral fellow, a PhD student and a technician, together with funds for consumables and equipment. For further information contact Prof. Dr. Marino Zerial by email ( zerial@mpi- cbg.de ). The Max Planck Society is committed to employ more disabled persons. The application of disabled persons is strongly encouraged. The Max Planck Society is committed to increase the number of female scientists. Women are particularly encouraged to apply. Please send your CV, publication list and a short description of research accomplishments and future plans to the address below by October 3, 2008. Two letters of recommendation should be sent separately by the application deadline to Personnel Department Code: 2008-RGL-1040@mpi-cbg.de Max Planck Institute of Molecular Cell Biology and Genetics Pfotenhauerstr. 108, 01307 Dresden, Germany Or e-mail: 2008-RGL-1040@mpi-cbg.de MAX-PLANCK-GESELLSCHAFT From xiaw from umbi.umd.edu Mon Oct 13 09:48:07 2008 From: xiaw from umbi.umd.edu (xiawei) Date: Mon Oct 13 10:43:23 2008 Subject: [Zbrafish] Help with microRNA fluorescent whole mount in situ hybridiztion Message-ID: Thanks everybody in advance for your help. Need a sensitive fluorescent method to detect microRNA expression in 36 hour old zebrafish embryos. Will LNA-modified probes labeled with Dig and detected by fluorescence labeled anti-dig work? Thanks a lot. Wei -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20081013/9806f2a5/attachment.html From Kevin_Kotredes from DFCI.HARVARD.EDU Thu Oct 16 13:08:08 2008 From: Kevin_Kotredes from DFCI.HARVARD.EDU (Kotredes, Kevin) Date: Thu Oct 16 13:12:29 2008 Subject: [Zbrafish] Zebrafish sperm cryopreservation Message-ID: I'm interested in learning and practicing the procedure to prepare zebrafish sperm for cryopreservation and later use for in vitro fertilization. Does anybody have a useful protocol they are willing to share or a resource that they can suggest? Thank you very much! Kevin P Kotredes Research Technician Look Zebrafish Lab Jimmy Fund Building, RM G03 Dana-Farber Cancer Institute Phone: 617-632-3545 Email: Kevin_Kotredes@dfci.harvard.edu The information transmitted in this electronic communication is intended only for the person or entity to whom it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon this information by persons or entities other than the intended recipient is prohibited. If you received this information in error, please contact the Compliance HelpLine at 800-856-1983 and properly dispose of this information. From driever from biologie.uni-freiburg.de Fri Oct 17 06:44:25 2008 From: driever from biologie.uni-freiburg.de (Wolfgang Driever) Date: Fri Oct 17 10:58:40 2008 Subject: [Zbrafish] Junior Research Group Leader Position Freiburg Message-ID: <091522DE-1FC8-45AF-AC9D-1E40315E84FF@biologie.uni-freiburg.de> Independent Junior Research Group Leader with tenure track option SIGNALLING RESEARCH AND ORGANOGENESIS The Centre for Biological Signalling Studies (bioss) and the Faculty of Biology at the University of Freiburg invite visionary scientists to apply for this position. The “Excellence Cluster” bioss aims at understanding cellular signalling processes that shape cells and organize them spatially into organs. To complement these efforts, we seek a signalling scientist with backgrounds ranging from 3D tissue culture to the analysis of mechanisms that control organogenesis. We expect an active integration into the bioss research environment (www.bioss.uni-freiburg.de). The position comes with an attractive start-up package. The initial appointment as W1-Professor will be for 4 years and can be extended for 2 more years. In case of outstanding results the possibility of a tenure track position (W3-Professorship) at the Faculty of Biology will be offered. The University of Freiburg is an equal opportunity employer. Applications of women are strongly encouraged. Handicapped candidates with equivalent qualifications will be given preference. The deadline for receipt of applications is 09.Novemer.2008. Applicants should use the application form (www.bioss.uni-freiburg.de/ positions/juniorleader). Applications including necessary supporting documents should be sent (in electronic form) to the administration of the bioss Excellence Cluster: Email: kontakt@bioss.uni-freiburg.de ******************************************************* Prof. Dr. Wolfgang Driever Developmental Biology Unit Department of Biology I University of Freiburg Hauptstrasse 1 D-79104 Freiburg GERMANY Tel: internat.: (49)-761 203 2587 admin. office. -2588 Fax: internat.: (49)-761 203 2597 email: driever@biologie.uni-freiburg.de http://www.biologie.uni-freiburg.de/data/bio1/driever/index.html ********************************************************* -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20081017/62d1750c/attachment.html From karlstrom from bio.umass.edu Fri Oct 17 12:35:39 2008 From: karlstrom from bio.umass.edu (rolfk) Date: Fri Oct 17 12:39:33 2008 Subject: [Zbrafish] Biology Chair Search at UMass Amherst Message-ID: <00364dd6-0a90-4f2f-8acc-e179dfe2c260@q35g2000hsg.googlegroups.com> Hi All, I wanted to spread the word that we are searching for a Department Chair at the University of Massachusetts, Amherst. The position will come with significant support and some future hires. The ad is copied below, feel free email me if there are any questions. Thanks, Rolf The Department of Biology at the University of Massachusetts at Amherst [website: www.bio.umass.edu/biology] seeks a senior level biologist with a nationally/internationally recognized active research program in any area of the life sciences, as a tenured full professor and department chair. Selection will be based on the combination of strength of research activity, fit to departmental needs, and experience and interest in administration. The University of Massachusetts has a strong commitment to expanding and strengthening research and training in the life sciences. These efforts are bolstered by the Commonwealth of Massachusetts $1 billion Life Sciences Initiative that includes increased faculty recruitment and capital expansion on the Amherst campus. The Department of Biology plays a key role in integrating life sciences activities across the UMass Amherst campus and provides leadership for four interdisciplinary life sciences graduate programs: Molecular and Cellular Biology [www.bio.umass.edu/mcb/], Neuroscience and Behavior [www.umass.edu/neuro/], Organismic and Evolutionary Biology [http:// www.bio.umass.edu/oeb/], and Plant Biology [www.bio.umass.edu/ plantbio/]. Evaluation of applications will begin on October 15, 2008 and continue until the position is filled. Applications should include a candidate’s CV and a statement of research interests, teaching philosophy and departmental administrative vision. Candidates will be contacted by the search committee to arrange for at least three letters of recommendation after the initial evaluation of applications is completed. Applications should be sent by email to: Biology Chair Search, c/o Ms. Cheryl Daggett, Personnel Manager, Dean of the College of Natural Sciences and Mathematics, Lederle Graduate Research Tower- Room 716, University of Massachusetts, Amherst, MA 01003 [email: daggett@nsm.umass.edu]. Please include Biology Chair Search in the subject line. The University of Massachusetts is an Equal Opportunity/Affirmative Action Employer. Women and members of minority groups are encouraged to apply. The Biology Department is aggressive in its efforts to hire candidates who will enhance the diversity and general balance of the faculty and the sciences. From finchg from ohsu.edu Fri Oct 17 18:26:56 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Fri Oct 17 18:28:37 2008 Subject: [Zbrafish] Re: Zebrafish sperm cryopreservation References: Message-ID: <8f2ce0db-4e32-4366-a8ba-843352838748@z6g2000pre.googlegroups.com> On Oct 16, 11:08?am, "Kotredes, Kevin" wrote: > I'm interested in learning and practicing the procedure to prepare zebrafish > sperm for cryopreservation and later use for in vitro fertilization. ?Does > anybody have a useful protocol they are willing to share or a resource that they > can suggest? ?Thank you very much! > > Kevin P Kotredes > Research Technician > Look Zebrafish Lab > Jimmy Fund Building, RM G03 > Dana-Farber Cancer Institute > Phone: 617-632-3545 > Email: Kevin_Kotre...@dfci.harvard.edu > > The information transmitted in this electronic communication is intended only > for the person or entity to whom it is addressed and may contain confidential > and/or privileged material. Any review, retransmission, dissemination or other > use of or taking of any action in reliance upon this information by persons or > entities other than the intended recipient is prohibited. If you received this > information in error, please contact the Compliance HelpLine at 800-856-1983 and > properly dispose of this information. Kevin, I'm trying to start up sperm cryopreservation at my lab as well. Multiple sperm freeze protocols can be found at the ZIRC site: http://zebrafish.org/zirc/documents/protocols.php Also, there is a protocol in the Christiane Nusslein-Volhard/Ralf Dahm Zfish book: http://books.google.com/books?id=zS-5CKOjCIgC&pg=PA30&dq=zebrafish+sperm+cryopreservation The Draper protocol appears to be the most widely used (e.g. at ZIRC, in TILLING at Moens Lab, etc.) and is well written/easy to follow. ....however, I am following it and thus far have been having a really difficult time getting the 1-3ul of sperm that the protocol recommends. If anyone reading this can recommend or describe a squeezing technique that can produce larger volumes, please please take the time to post some advice. I have tried everything I can think of, variety of forceps, fingers, variety of pressures, variety of anatomical regions, sexy music, etc. Anyone? Please........ From castrand from mail.nih.gov Mon Oct 20 08:24:19 2008 From: castrand from mail.nih.gov (Daniel Castranova) Date: Mon Oct 20 11:41:13 2008 Subject: [Zbrafish] Re: Zebrafish sperm cryopreservation In-Reply-To: <8f2ce0db-4e32-4366-a8ba-843352838748@z6g2000pre.googlegroups.com> References: <8f2ce0db-4e32-4366-a8ba-843352838748@z6g2000pre.googlegroups.com> Message-ID: <495FD9A7-D0B6-430B-9CE6-C3E3637C97A4@mail.nih.gov> Hello Kevin and others, I have been doing zebrafish sperm cryopreservation for a while and I agree, the 1-3ul of sperm described in many protocols isn't realistic. I find that I get between 0.5 and 2ul with the average around 0.75 - 1ul. I don't think that there is an extraction technique that will help increase your volume, but I don't think you should be that concerned about it. I have fertilized over 800 eggs with 0.5ul of sperm (fresh not frozen) so I think there is enough sperm in small samples to get the job done, it is a matter of properly freezing and thawing the sample. I have had the most success with a modified version of the protocol found in the zebrafish book. I cut capillary tubes to 1" and freeze the tubes inside cryovials to save space in our N2 dewar. I also use 10ul of freezing media. If anyone would like a copy of our detailed protocol contact me off of the list and I will send it out. I think the key to successful sperm freezing is practice. It took me about a year before I was happy with my results and I spent a lot of time trying various protocols that I could never get to work. I hope that helps, Dan Daniel Castranova Aquatic Specialist Charles River Laboratories - Contractor Unit on Vertebrate Organogenesis Laboratory of Molecular Genetics, NICHD, NIH Building 6B, Room 322, 6 Center Drive Bethesda, MD 20892 (301)594-0904 castrand@mail.nih.gov On Oct 17, 2008, at 7:26 PM, finchg@ohsu.edu wrote: > On Oct 16, 11:08 am, "Kotredes, Kevin" > wrote: >> I'm interested in learning and practicing the procedure to prepare >> zebrafish >> sperm for cryopreservation and later use for in vitro >> fertilization. Does >> anybody have a useful protocol they are willing to share or a >> resource that they >> can suggest? Thank you very much! >> >> Kevin P Kotredes >> Research Technician >> Look Zebrafish Lab >> Jimmy Fund Building, RM G03 >> Dana-Farber Cancer Institute >> Phone: 617-632-3545 >> Email: Kevin_Kotre...@dfci.harvard.edu >> >> The information transmitted in this electronic communication is >> intended only >> for the person or entity to whom it is addressed and may contain >> confidential >> and/or privileged material. Any review, retransmission, >> dissemination or other >> use of or taking of any action in reliance upon this information by >> persons or >> entities other than the intended recipient is prohibited. If you >> received this >> information in error, please contact the Compliance HelpLine at >> 800-856-1983 and >> properly dispose of this information. > > Kevin, > I'm trying to start up sperm cryopreservation at my lab as well. > Multiple sperm freeze protocols can be found at the ZIRC site: > http://zebrafish.org/zirc/documents/protocols.php > Also, there is a protocol in the Christiane Nusslein-Volhard/Ralf Dahm > Zfish book: > http://books.google.com/books?id=zS-5CKOjCIgC&pg=PA30&dq=zebrafish+sperm+cryopreservation > The Draper protocol appears to be the most widely used (e.g. at ZIRC, > in TILLING at Moens Lab, etc.) and is well written/easy to follow. > > ....however, I am following it and thus far have been having a really > difficult time getting the 1-3ul of sperm that the protocol > recommends. If anyone reading this can recommend or describe a > squeezing technique that can produce larger volumes, please please > take the time to post some advice. I have tried everything I can > think of, variety of forceps, fingers, variety of pressures, variety > of anatomical regions, sexy music, etc. Anyone? Please........ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From herbomel from pasteur.fr Tue Oct 21 10:22:20 2008 From: herbomel from pasteur.fr (Philippe Herbomel) Date: Tue Oct 21 11:59:11 2008 Subject: [Zbrafish] td-tomato vs mcherry as a red fluo reporter Message-ID: Dear All, We are wondering about the best red fluorescent reporter to make transgenic zebrafish with. If one does not want to target the protein to a particular compartment (membrane or nucleus), DsRed2, the most used presently, is probably the best (very bright, with reasonable maturation time). If otherwise, the most used presently is mCherry, which is quite (4- fold) less bright. Now from the Tsien lab papers' data, td-Tomato appears both much (over 5-fold) brighter than mCherry, and still compatible with membrane or nuclear targeting; and indeed at least two papers then provided evidence that td-tomato was not only far superior to mcherry but even to GFP (in terms of brightness) as a reporter in transgenic mice. Now surprisingly, when a colleague of us injected either mcherry or tdtomato mRNA into 1-cell zebrafish embryos, she found td-Tomato to be less bright than mcherry. So my question is: has anyone found this too, and/or tried to make transgenic zebrafish with td-Tomato as reporter ? And what about its actual brightness (and decay upon illumination) in zebrafish embryos/ larvae, as compared to the mcherry and dsred2 ? Thanks in advance for your testimonies on this. I will post a summary of your responses ! Philippe Herbomel ––––––––––––––––––––––––––––––––––––––––– Philippe Herbomel Unite Macrophages et Developpement de l'Immunite Departement de Biologie du Developpement Institut Pasteur 25 rue du Dr Roux, 75724 Paris cedex 15, France tel. : 33 1 44 38 95 29 mobile: 33 6 73 35 74 20 fax : 33 1 45 68 89 21 http://www.pasteur.fr/recherche/RAR/RAR2006/Mdi-en.html –––––––––––––––––––––––––––––––––––––– Change the world one loan at a time - visit Kiva.org to find out how -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20081021/d3d85980/attachment.html From herbomel from pasteur.fr Tue Oct 21 10:22:20 2008 From: herbomel from pasteur.fr (Philippe Herbomel) Date: Wed Oct 22 11:56:04 2008 Subject: [Zbrafish] td-tomato vs mcherry as a red fluo reporter Message-ID: Dear All, We are wondering about the best red fluorescent reporter to make transgenic zebrafish with. If one does not want to target the protein to a particular compartment (membrane or nucleus), DsRed2, the most used presently, is probably the best (very bright, with reasonable maturation time). If otherwise, the most used presently is mCherry, which is quite (4- fold) less bright. Now from the Tsien lab papers' data, td-Tomato appears both much (over 5-fold) brighter than mCherry, and still compatible with membrane or nuclear targeting; and indeed at least two papers then provided evidence that td-tomato was not only far superior to mcherry but even to GFP (in terms of brightness) as a reporter in transgenic mice. Now surprisingly, when a colleague of us injected either mcherry or tdtomato mRNA into 1-cell zebrafish embryos, she found td-Tomato to be less bright than mcherry. So my question is: has anyone found this too, and/or tried to make transgenic zebrafish with td-Tomato as reporter ? And what about its actual brightness (and decay upon illumination) in zebrafish embryos/ larvae, as compared to the mcherry and dsred2 ? Thanks in advance for your testimonies on this. I will post a summary of your responses ! Philippe Herbomel ––––––––––––––––––––––––––––––––––––––––– Philippe Herbomel Unite Macrophages et Developpement de l'Immunite Departement de Biologie du Developpement Institut Pasteur 25 rue du Dr Roux, 75724 Paris cedex 15, France tel. : 33 1 44 38 95 29 mobile: 33 6 73 35 74 20 fax : 33 1 45 68 89 21 http://www.pasteur.fr/recherche/RAR/RAR2006/Mdi-en.html –––––––––––––––––––––––––––––––––––––– Change the world one loan at a time - visit Kiva.org to find out how -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20081021/d3d85980/attachment-0002.html