From claudia_hohn from hotmail.com Sat Aug 2 14:13:35 2008 From: claudia_hohn from hotmail.com (Claudia) Date: Mon Aug 4 11:31:27 2008 Subject: [Zbrafish] Re: help with parasite infecting embryos References: Message-ID: <07c408a1-99bb-44c0-b80c-6fb71dc85272@x35g2000hsb.googlegroups.com> On Jul 31, 5:47 pm, "David G. White" wrote: > The little filaments are most likely fungal hyphae, saprolegnia etc and would come more problematic with a broken UV. One method of treating the embryos would be using methylene blue, which can be found at any pet store. Add enough to your egg medium until it turns blue. There is a few draw backs to methylene blue. It will kill bacterial beds so do not want to use it on any kind of water recirculating nursery with a biofilter. Also it can stain cells blue. The fungal spores will exist in your fish system water even with a fully-functioning UV. > > If you use spawning cages you can try setting up your fish later in the day, so that you reduce the amount of fecal matter deposited in the cages with the eggs. We collect the eggs promptly in the morning. Pour the eggs through a 200 um screen and rinse them with fresh egg water in to a petri dish. Under the scope remove all debris and any infertile eggs. Good eggs should be perfectly round with a dark round yolk. If your not certain if the egg is good or not I would remove it because a few bad eggs can spoil the whole lot. The next day look over the eggs again, and clean if necessary. We do not use methylene blue. > > You also might want to increase maintenance on your UV, replacing the bulb and cleaning the quartz sleeve or perhaps up-sizing it. > > Good luck > > David G White > Research Coordinator > H225 Zebrafish Lab > University of Washington > Department of Biological Structure > HSB G514 Box 357420 > 1959 NE Pacific Street > Seattle, WA 98195-7420 > Tel. 206-685-7512 > FAX 206-543-1524 > > On Thu, 31 Jul 2008, lampe1 wrote: > > Hello! > > > In our zebrafish facility, we frequently have embryos infected with > > some sort of parasite. It looks like little filaments of cotton which > > surround the embryo. When this happens, the embryos usually die within > > 24-72 hours. Usually bleaching (according to ZFIN protocol--0.1mL 5% > > sodium hypochlorite in 170mL system water, bleach 5 min, system water > > 5 min, bleach 5 min, rinse 2+ times in system water) and replacing the > > water stops this and saves the embryos. > > > Recently, this parasite has been worse (likely due to our UV > > sterilizer being broken for ~1 week). One person in the lab bleached > > the embryos and rinsed them several times and yet they still all died > > and were infected with this parasite. > > > Does anyone have any idea what this parasite may be? I am having > > trouble since I have no idea what kind of organism it even is. Does > > anyone have any ideas for other ways to bleach/otherwise save embryos? > > > I appreciate any help! > > > Thanks, > > > Rebecca Lampe > > Fish Care Technician > > UMBC Department of Biology > > lam...@umbc.edu > > > _______________________________________________ > > Zbrafish mailing list > > Zbraf...@net.bio.net > >http://www.bio.net/biomail/listinfo/zbrafish Hi, i had this problem before. I think it has less to do with egg disinfection..... What I found was the source of contamination was the paramecium infusion I was feeding. After I discarded all paramecium, disinfected all containers and ordered a new starter culture, the problem disappeared. I also take more care now of the paramecium cultures see my protocol: http://www.cvm.msstate.edu/zebrafish/index.html Hope this helps, Claudia From ian.bessell from zeiglerfeed.com Fri Aug 1 16:03:40 2008 From: ian.bessell from zeiglerfeed.com (Ian Bessell) Date: Mon Aug 4 11:31:41 2008 Subject: [Zbrafish] Zebrafish food In-Reply-To: <1D117326B8F30943ACD9A47E1BEE2941C4B57F@CHEXV2.CHBOSTON.ORG> References: <1D117326B8F30943ACD9A47E1BEE2941C4B57F@CHEXV2.CHBOSTON.ORG> Message-ID: <32B0615DEB9B4F439CAE7F94CFA0E7B9012CB232@SRVGARMX.ZEIGLER.LCL> Hi Sarah, Most processed feeds have a shelf-life of about 3-6 months without any additional preservation depending on the formulation and packaging. If a product is packaged under nitrogen or vacuum sealed, or contains extra preservatives the shelf-life may be longer. Refrigeration and/or freezing food can also extend the shelf-life. Lastly, irradiation can be used to also extend the shelf-life of a product. If the feed you have at your facility is 5 years old chances are that it no longer meets the minimum guarantees on the tag and may be providing inadequate nutrition for your fish. It is hard to say for sure, but I think it is a safe guess at this point. Zeigler Brother's offers an adult Zebrafish diet through Aquatic Habitats Inc. Please visit http://www.zeiglerfeed.com/product_literature/lab%20research%20literatur e_Aquatic/Adult%20Zebrafish%20Diet.pdf for additional information about this product. We also offer several larval diets that can be used for larval zebrafish, also available through Aquatic-Habitats. If you need additional information, please just let me know. Best regards, Ian ________________________________________ Ian S. Bessell, M.A.B. Sales Specialist-Pet, Zoo and Lab Diets Zeigler Bros.Feed, Inc. 717-968-6915 cell 786-242-7157 Florida office 786-242-6549 fax Skype Contact: ian.bessell Ian.Bessell@zeiglerfeed.com www.zeiglerfeed.com www.monsterdiets.com _______________________________________ CONFIDENTIALITY NOTICE: The information contained in this document from Zeigler Bros., Inc. is privileged and confidential and is intended for the sole use of the person(s) or entities named on the cover letter/fax/memo/e-mail. If you are not an intended recipient of this document, the dissemination, distribution, copying or use of the information it contains is strictly prohibited, please contact the sender, and delete or discard the material. -----Original Message----- From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Lawrence, Christian Sent: Thursday, July 31, 2008 3:58 PM To: SarBOgden@gmail.com; bionet-organisms-zebrafish@moderators.isc.org Subject: Re: [Zbrafish] Zebrafish food The shelf life on processed feeds is generally no more than 3 months, if they are stored correctly (cool and dry). If you don't store properly, it is less than that. There are several processed feeds out there that support good production for zebrafish, although none have been demonstrated to be nutritionally complete. We use Lansy NRD 400-600 pellet by INVE aquaculture. This works best when used as part of a diet that includes Artemia. It costs approximately 7-10$/kg. Chris ----- Original Message ----- From: zbrafish-bounces@oat.bio.indiana.edu To: bionet-organisms-zebrafish@moderators.isc.org Sent: Thu Jul 31 15:15:00 2008 Subject: [Zbrafish] Zebrafish food Hello, In my facility we've been feeding Rangen Trout and Salmon starter. We've been using the same bag that we've kept in the fridge for 5 years. Does anyone use this food and know if it lasts this long? We are looking to order new food but are looking for other options. Any suggestions would be greatly appreciated. Thanks, Sarah Lab Technician UMBC sarbogden@gmail.com _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish 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. _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From noopur.mandrekar from gmail.com Fri Aug 1 06:19:53 2008 From: noopur.mandrekar from gmail.com (Noopur) Date: Mon Aug 4 11:32:53 2008 Subject: [Zbrafish] Re: Zebrafish training References: <200807281703.m6SH35O27581@net.bio.net> Message-ID: Hi Caroline, I am Noopur from India. I am interested in learning few techniques like microinjection and in-situ hybridization. Do you offer such kind of a training? Are there any workshops conducted which include screening assays or these techniques? Thanking You in advance Best Wishes & Regards Noopur Mandrekar On Jul 30, 2:46?pm, Caroline Parkin wrote: > Hi, > Please can you tell us where you are located? Within the UK/Europe it ? > may be possible to get training here at the University of Sheffield ? > (this is something new we are considering offering, depending on the ? > amount of interest - feedback on this idea would be welcome). > best wishes, > Caroline > caroline.par...@shef.ac.uk > > On 28 Jul 2008, at 18:03, zbrafish-requ...@oat.bio.indiana.edu wrote: > > > > > Send Zbrafish mailing list submissions to > > ? ?zbraf...@net.bio.net > > > To subscribe or unsubscribe via the World Wide Web, visit > > ? ?http://www.bio.net/biomail/listinfo/zbrafish > > or, via email, send a message with subject or body 'help' to > > ? ?zbrafish-requ...@net.bio.net > > > You can reach the person managing the list at > > ? ?zbrafish-ow...@net.bio.net > > > When replying, please edit your Subject line so it is more specific > > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > > ? 1. Zebrafish training session (movingfish) > > > ---------------------------------------------------------------------- > > > Message: 1 > > Date: Fri, 25 Jul 2008 23:26:02 -0700 (PDT) > > From: movingfish > > Subject: [Zbrafish] Zebrafish training session > > To: bionet-organisms-zebraf...@moderators.isc.org > > Message-ID: > > ? ? > > Content-Type: text/plain; charset=ISO-8859-1 > > > Does anyone know where i can get basic training on zebrafish > > development and experimental protocol? > > > ------------------------------ > > > _______________________________________________ > > Zbrafish mailing list > > Zbraf...@net.bio.net > >http://www.bio.net/biomail/listinfo/zbrafish > > > End of Zbrafish Digest, Vol 38, Issue 16 > > ****************************************- Hide quoted text - > > - Show quoted text - From weibinz from umich.edu Fri Aug 1 12:46:57 2008 From: weibinz from umich.edu (Weibin Zhou) Date: Mon Aug 4 11:34:42 2008 Subject: [Zbrafish] GFP and in situ Message-ID: <20080801134657.25997h14wd85hoo4@web.mail.umich.edu> Hi, I am wondering whether it is possible to preserve GFP fluorescence after in situ hybridization (whole-mount or sectioned). If you happen to have a protocol that works, please let me know. Thank you very much. Weibin From finchg from ohsu.edu Mon Aug 4 12:25:15 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Mon Aug 4 12:35:31 2008 Subject: [Zbrafish] Re: Swarming zebrafish embryo parasites References: Message-ID: On Jul 31, 6:16 pm, RachelRay...@gmail.com wrote: > While performing maintenance on a clutch of 48 hour zebrafish embryos, > I came across one with a swarm of parasites within its chorion. All > the other embryos in the clutch were fine. I examined the embryo 5 > hours later and it was almost completely devoured. I assume that this > is a parasite that the mother passed on? Unfortunately, the cross was > albino stock and the individual fish was put back into a group tank > after breeding. What were those things and what action if any needs > to be taken? > > If anyone would like a picture or video of the embryo I’d be happy to > send one in hopes of identifying the parasite. See previous discussion on this board concerned with "coleps" for the most information. Also: http://zfin.org/zf_info/monitor/vol5.1/vol5.1.html#Coleps,%20Scourge%20of%20the%20Baby%20Zebrafish These commonly contaminate embryo dishes and are frequently found inside the chorion. There is no silver bullet, if you have adults on a seperate system from your fry you should try fluctuating the conductivity significantly in the adult system on a semi-regular basis. Clean "bio-sludge" from system drain gutters and spillways and frequently ethanol work areas, incubators, etc. Bleach all eggs as a short-term control. Perhaps the most important single factor: thouroghly clean your embryo dishes of everything but the embryos on a daily basis (leftover chorion is the colep equivelant of the deluxe nacho platter at T.G.I.Fridays). From hmcallis from Princeton.EDU Mon Aug 4 14:35:32 2008 From: hmcallis from Princeton.EDU (Heather S McAllister (hmcallis@Princeton.EDU)) Date: Mon Aug 4 14:53:03 2008 Subject: [Zbrafish] tricaine use before RT-PCR Message-ID: A member of our lab is wondering if using tricaine on fish from 7dpf to 60 dpf before putting them in trizol will affect the results of an RT-PCR. Any help would be appreciated. Heather McAllister Research Specialist II Burdine Lab Department of Molecular Biology Princeton University Princeton, NJ 08544 hmcallis@princeton.edu 609-258-5782 From dane from jhmi.edu Mon Aug 4 15:18:40 2008 From: dane from jhmi.edu (Dane Witmer) Date: Mon Aug 4 15:30:53 2008 Subject: [Zbrafish] monolith (mnl) mutant Message-ID: Dear List, Is there any mapping info available for this mutant? I have been unable to find anything. Thanks! Dane Witmer dane@jhmi.edu -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080804/62aa990b/attachment.html From april from zebrafish.org Mon Aug 4 17:03:36 2008 From: april from zebrafish.org (April) Date: Mon Aug 4 17:05:25 2008 Subject: [Zbrafish] Swarming zebrafish embryo parasites In-Reply-To: <9d97a969-187c-46c4-a594-5ea426f8b81f@56g2000hsm.googlegroups.com> References: <9d97a969-187c-46c4-a594-5ea426f8b81f@56g2000hsm.googlegroups.com> Message-ID: <55F16746-B86B-4B6D-A242-2FF54D43C93E@zebrafish.org> Hi Rachel, To determine if these are in fact coleps, look at them under a compound microscope. Coleps are very easy to identify. They move in a swirling pattern and their body's are covered in plates that make them look like hand grenades. Unhatched or recently hatch embryos that have not developed swim bladders are the most susceptible to coleps. A swarm of coleps can consume an entire embryo within 45 minutes. Once embryos are swimming they can fend them off quite easily. If the parasites were only found in 1 embryo, I wouldn't worry to much yet. Keep your embryos as clean as possible and remove anything suspicious. You should also consider bleaching your embryos, if you are not currently. I would be happy to take a look at some pictures and help identify the parasite. Good luck, and let me know if you have any questions. April On Jul 31, 2008, at 6:16 PM, RachelRaynes@gmail.com wrote: > While performing maintenance on a clutch of 48 hour zebrafish embryos, > I came across one with a swarm of parasites within its chorion. All > the other embryos in the clutch were fine. I examined the embryo 5 > hours later and it was almost completely devoured. I assume that this > is a parasite that the mother passed on? Unfortunately, the cross was > albino stock and the individual fish was put back into a group tank > after breeding. What were those things and what action if any needs > to be taken? > > If anyone would like a picture or video of the embryo I’d be happy to > send one in hopes of identifying the parasite. > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish ----------------------- April R. Freeman (Mazanec) ZIRC Manager Zebrafish Int.'l Resource Center 5274 University of Oregon Eugene, OR 97403-5274 Phone: (541) 346-6028 ext. 24 Fax: (541) 346-6151 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080804/2182262d/attachment.html From chi-bin.chien from neuro.utah.edu Mon Aug 4 16:54:30 2008 From: chi-bin.chien from neuro.utah.edu (Chi-Bin Chien) Date: Mon Aug 4 18:07:26 2008 Subject: [Zbrafish] GFP and in situ In-Reply-To: <20080801134657.25997h14wd85hoo4@web.mail.umich.edu> References: <20080801134657.25997h14wd85hoo4@web.mail.umich.edu> Message-ID: Native GFP fluorescence is sensitive to pH and organic solvents, and does not survive in situ protocols. However, in our hands anti-GFP antibody staining after in situs works very well. See the following reference: http://www.jneurosci.org/cgi/content/full/26/51/13328 Chi-Bin Chien At 1:46 PM -0400 8/1/08, Weibin Zhou wrote: >Hi, >I am wondering whether it is possible to preserve GFP fluorescence >after in situ hybridization (whole-mount or sectioned). If you >happen to have a protocol that works, please let me know. Thank you >very much. > >Weibin > > > >_______________________________________________ >Zbrafish mailing list >Zbrafish@net.bio.net >http://www.bio.net/biomail/listinfo/zbrafish From yang.x from neu.edu Tue Aug 5 11:08:38 2008 From: yang.x from neu.edu (yang.x@neu.edu) Date: Tue Aug 5 11:38:13 2008 Subject: [Zbrafish] needle calibration Message-ID: Hi, I just started to do microinjection. I am wondering how to calibrate the needles so that I know the amount I am injecting. Thanks a lot! From jmoulton from gene-tools.com Tue Aug 5 12:03:02 2008 From: jmoulton from gene-tools.com (Jon D.Moulton) Date: Tue Aug 5 12:09:41 2008 Subject: [Zbrafish] needle calibration In-Reply-To: References: Message-ID: <489887C6.80401@gene-tools.com> Regarding the question: "Hi, I just started to do microinjection. I am wondering how to calibrate the needles so that I know the amount I am injecting." See the supporting online material of the following citation for *methods for characterizing microinjected volume* per embryo. Riedel-Kruse IH, Muller C, Oates AC. Synchrony dynamics during initiation, failure, and rescue of the segmentation clock. Science. 2007 Sep 28;317(5846):1911-5. Epub 2007 Aug 16. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=17702912 http://www.sciencemag.org/cgi/data/1142538/DC1/1 Best regards, - Jon Jon D. Moulton, Ph.D. Diagnostics and Special Projects GENE TOOLS, LLC jmoulton@gene-tools.com http://network.nature.com/group/morpholinos yang.x@neu.edu wrote: > Hi, I just started to do microinjection. I am wondering how to > calibrate the needles so that I know the amount I am injecting. Thanks > a lot! > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080805/eae4014a/attachment.html From yang.x from neu.edu Tue Aug 5 16:11:59 2008 From: yang.x from neu.edu (yang.x@neu.edu) Date: Tue Aug 5 16:29:38 2008 Subject: [Zbrafish] storage of RNA samples for injection Message-ID: <9c025b2b-38b3-493f-af8d-0a9a11b1d7ae@m73g2000hsh.googlegroups.com> Just got another question. If I am injecting mRNA to embryos, shall I leave the left diluted mRNA at 4 degree for the next day? How long would it be good? Thanks a lot! From dcb1 from hi.is Tue Aug 5 14:59:58 2008 From: dcb1 from hi.is (Daniela Carmen Broekman) Date: Wed Aug 6 11:58:59 2008 Subject: [Zbrafish] Cell line constitutive expression Message-ID: <58646.130.208.165.5.1217966398.squirrel@webmail.hi.is> Hi, I'm working with a Salmonid cell line and started having a problem with constititive expression of my target gene although my work depends on expression being inducible. Does anyone have an idea why a cell line suddenly starts to express constitutively and does not recover from it? -- _________________________________________________ ><((((?> From rburdine from Princeton.EDU Tue Aug 5 19:51:34 2008 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Wed Aug 6 11:59:19 2008 Subject: [Zbrafish] storage of RNA samples for injection References: <9c025b2b-38b3-493f-af8d-0a9a11b1d7ae@m73g2000hsh.googlegroups.com> Message-ID: <04AF14C4EF3CF34682460FB1FDBA98C02B45F3@MBCLUSTER.pu.win.princeton.edu> In general, We make small aliquots of RNA (5-10ul) diluted to the correct concentration in buffer/phenol red and store them at -80C. If we need to determine the correct concentration to inject, we store small aliquots of concentrated RNA at -80C and dilute into buffer as needed. When we thaw an aliquot of RNA diluted in buffer to inject, we toss whatever we don't use. I would be surprised after going in and out of a tube of RNA and leaving it at 4degrees if there would be anything left the next day. Even if it didn't completely degrade, the concentration would be different for sure and may lead to variability in your experiments. Just my 2 cents worth, Becky --------------------------------------------------- Rebecca D. Burdine, Ph.D. Assistant Professor Dept. of Molecular Biology Princeton University Washington Road Mof 433 Princeton, NJ 08544 Phone: (609) 258-7515 Fax: (609) 258-1343 Email: rburdine@princeton.edu Admin Assistant: Cathy Falk (609) 258-1604 ________________________________ From: zbrafish-bounces@oat.bio.indiana.edu on behalf of yang.x@neu.edu Sent: Tue 8/5/2008 5:11 PM To: bionet-organisms-zebrafish@moderators.isc.org Subject: [Zbrafish] storage of RNA samples for injection Just got another question. If I am injecting mRNA to embryos, shall I leave the left diluted mRNA at 4 degree for the next day? How long would it be good? Thanks a lot! _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080805/9702003f/attachment.html From tgreiling from gmail.com Tue Aug 5 19:47:59 2008 From: tgreiling from gmail.com (tgreiling@gmail.com) Date: Wed Aug 6 11:59:40 2008 Subject: [Zbrafish] IHC membrane stain? Message-ID: <3d923ad2-8ec5-4c54-a1e3-19c4ee619a60@b2g2000prf.googlegroups.com> Does anyone know of a good cell membrane antibody or lectin for counterstaining frozen sections of zebrafish embryos? I use wheat germ agglutinin on mouse sections and it gives beautiful membrane outlines but it didn't recognize anything in fish. Ideas? From braubach from dal.ca Thu Aug 7 14:02:16 2008 From: braubach from dal.ca (Oliver Braubach) Date: Thu Aug 7 14:05:57 2008 Subject: [Zbrafish] Zebrafish Age / Size Variations Message-ID: <086CBF4D-3A7D-44C4-8F33-2D3B0E8D977A@dal.ca> Dear All. I have been working on aspects of postembryonic development in the zebrafish brain, and have been trying to determine a standard method for 'aging' zebrafish. As many of you may be aware, the development of individual fish varies greatly during the first month of development, which ultimately leads to very noticeable differences in the size of individual fish and their organs. Thus, age does not accurately characterize developmental stages of fish, and I have been trying to determine if there's another standard method to do so. Has anyone tried to correlate body length to fish age? Are there any citations for expected / optimal size of fish are a certain age? Any input regarding this problem would be greatly appreciated. Best, Oliver Braubach Department of Physiology and Biophysics Dalhousie University Halifax, NS B3H 1X5 Canada Tel: 902.494.6471 Fax: 902.494.1685 From Thomas.Bartman from cchmc.org Thu Aug 7 14:44:01 2008 From: Thomas.Bartman from cchmc.org (Thomas Bartman) Date: Thu Aug 7 16:46:18 2008 Subject: [Zbrafish] Zebrafish Age / Size Variations Message-ID: <489B1841020000760001745C@n6mcgw16.cchmc.org> Oliver, I think this is an important issue which is sometimes overlooked. We are looking at late heart development, and are trying to get work published which shows that morphogenesis is tied strongly to body mass (therefore, theoretically physiological parameters) and not age. There was a poster at the zf meeting which proposed what seemed to be a nice method for staging larvae / young adults based on length and physical characteristics. I don't have the abstracts in front of me. Maybe the authors of that work are on this newsgroup and will chime in? Tom Bartman Cincinnati Children's Hospital Thomas Bartman, M.D., Ph.D. Cincinnati Children's Hospital Divisions of Neonatology, Pulmonary Biology and Developmental Biology 3333 Burnet Avenue Cincinnati, OH 45229 (513) 636-9902 >>> Oliver Braubach 08/07/08 3:02 PM >>> Dear All. I have been working on aspects of postembryonic development in the zebrafish brain, and have been trying to determine a standard method for 'aging' zebrafish. As many of you may be aware, the development of individual fish varies greatly during the first month of development, which ultimately leads to very noticeable differences in the size of individual fish and their organs. Thus, age does not accurately characterize developmental stages of fish, and I have been trying to determine if there's another standard method to do so. Has anyone tried to correlate body length to fish age? Are there any citations for expected / optimal size of fish are a certain age? Any input regarding this problem would be greatly appreciated. Best, Oliver Braubach Department of Physiology and Biophysics Dalhousie University Halifax, NS B3H 1X5 Canada Tel: 902.494.6471 Fax: 902.494.1685 _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From herbomel from pasteur.fr Fri Aug 8 06:12:00 2008 From: herbomel from pasteur.fr (Philippe Herbomel) Date: Fri Aug 8 11:10:41 2008 Subject: [Zbrafish] Zebrafish Age / Size Variations; In-Reply-To: <086CBF4D-3A7D-44C4-8F33-2D3B0E8D977A@dal.ca> References: <086CBF4D-3A7D-44C4-8F33-2D3B0E8D977A@dal.ca> Message-ID: Sorry, please replace my previous message (in which the reference of the book wasn't quite accurate) by this one. Philippe Herbomel Dear Oliver, Body length appears indeed to be the adequate, simple parameter for estimating postembryonic developmental stage. Our reference in this matter is Tom Schilling's excellent chapter on "The morphology of larval and adult zebrafish" (pp. 59-94 In "Zebrafish: A Practical Approach." 2002, eds. C. Nusslein-Volhard and R. Dahm, Oxford Univ. Press), in which Table 2 provides a typical correspondence between "age" and body length" up to 90 days. In their study of the development and maturation of the immune system in zebrafish, Lam et al. ( Dev. Compar. Immunol. 28 : 9-28, 2004) also described this maturation relative not only to age but also to body length (see Discussion), which is obviously more significant and helpful. You will see that they give the same correspondence between age and body length as Tom Schilling at 2 weeks but not at 4 weeks. Interestingly, an older paper comparing the role of thyroid hormone in zebrafish (vs. axolotl) metamorphosis, i.e. their gradual transformation form larvae into juveniles, also used body length to express the developmental stages (Brown D., PNAS 94 : 13011-16, 1997). This is obviously an important issue, and certainly all zebrafish studies dealing with stages past the first week of development should refer to body length in addition to or instead of nominal age. Also, the community should agree about how to measure that length. In Figure 1 and Table 2 of his chapter refered above, Tom Schilling provided two different measurements : 'Full length' i.e. up to the tip of the caudal fin, and 'standard length', ie up to the base of the caudal fin. I wasn't at the last Madison meeting but since there was a workshop there about "working with post-embryonic stages", maybe this staging issue was raised ? I would be interested to learn about this from participants to this workshop. Best wishes, Philippe ––––––––––––––––––––––––––––––––––––––––– 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 Le 7 août 08 à 21:02, Oliver Braubach a écrit : > Dear All. > > I have been working on aspects of postembryonic development in the > zebrafish brain, and have been trying to determine a standard > method for 'aging' zebrafish. As many of you may be aware, the > development of individual fish varies greatly during the first > month of development, which ultimately leads to very noticeable > differences in the size of individual fish and their organs. > > Thus, age does not accurately characterize developmental stages of > fish, and I have been trying to determine if there's another > standard method to do so. Has anyone tried to correlate body length > to fish age? Are there any citations for expected / optimal size of > fish are a certain age? > > Any input regarding this problem would be greatly appreciated. > Best, > > Oliver Braubach > Department of Physiology and Biophysics > Dalhousie University > Halifax, NS B3H 1X5 Canada > Tel: 902.494.6471 > Fax: 902.494.1685 > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080808/e568d415/attachment.html From tgenade from gmail.com Fri Aug 8 02:31:17 2008 From: tgenade from gmail.com (Tyrone) Date: Fri Aug 8 11:11:06 2008 Subject: [Zbrafish] Re: Zebrafish Age / Size Variations References: Message-ID: <0a714658-5464-4690-8ced-883f0a85f0af@b1g2000hsg.googlegroups.com> On Aug 7, 9:02?pm, Oliver Braubach wrote: > I have been working on aspects of postembryonic development in the ? > zebrafish brain, and have been trying to determine a standard method ? > for 'aging' zebrafish. As many of you may be aware, the development of ? > individual fish varies greatly during the first month of development, ? > which ultimately leads to very noticeable differences in the size of ? > individual fish and their organs. Hello Oliver, I suggest you track down the papers by Markofsky et al concerning his research on Nothobranchius guentheri. Growth is very important with aging. It is commonly observed that "runt" fishes tend to live longer, and thus, must age (accumulate aging-related pathologies) slower. Markofsky didn't solve the problem of different growth rate, he simply used a lot more specimens and let the stats even out the problem. You may wish to explore different rearing conditions. My experience as a aquarist has taught me that the larger the tank and the heavier the feeding the better the growth rate and the more even the growth rate in the brood. Another option is to decide on a size cut off per age. For instance, at day x you will only use "average +/- SD" size fish. This of course means you will have to do carefully controled experiments monitoring growth rate daily for a shoal of fry. This is easily acomplished with behavioural software such as "Ethovision", where the fish are placed in a tank that rests over some graph paper and the tank and fish photographed. The program does the hard work. This is briefly explained in Genade et al. Aging Cell. 4:223--233, 2005. Hope this is of some help. Kind regards From Christian.Lawrence from childrens.harvard.edu Fri Aug 8 11:54:41 2008 From: Christian.Lawrence from childrens.harvard.edu (Christian Lawrence) Date: Fri Aug 8 11:56:48 2008 Subject: [Zbrafish] Zebrafish Age / Size Variations; In-Reply-To: Message-ID: FYI: For discussion of relationships between between sexual differentiation, body length, and age, see: Maak H, Segner CR, and Tyler R (2003) Ontogeny of sexual differentiation in different strains of zebrafish (Danio rerio). Fish Physiology and Biochemistry, Volume 28, Numbers 1-4, 2003 , pp. 125-128(4). Chris On 8/8/08 7:12 AM, "Philippe Herbomel" wrote: > Sorry, please replace my previous message (in which the reference of the book > wasn't quite accurate) > by this one. > Philippe Herbomel > > > Dear Oliver, > > Body length appears indeed to be the adequate, simple parameter for estimating > postembryonic developmental stage. > Our reference in this matter is Tom Schilling's excellent chapter on "The > morphology of larval and adult zebrafish" (pp. 59-94?In "Zebrafish: A > Practical Approach." 2002, eds. C. Nusslein-Volhard and R. Dahm, Oxford Univ. > Press), in which Table 2 provides a typical?correspondence between "age" and > body length" up to 90 days. > In their study of the development and maturation of the immune system in > zebrafish, Lam et al. ( Dev. Compar. Immunol. 28 : 9-28, 2004) also described > this maturation relative?not only to age but also to body length (see > Discussion), which is obviously more significant and helpful. You will see > that they give the same correspondence between age and body length as Tom > Schilling at 2 weeks but not at 4 weeks. > Interestingly, an older paper comparing the role of thyroid hormone in > zebrafish (vs. axolotl) metamorphosis, i.e. their gradual transformation form > larvae into juveniles, also used body length to express the developmental > stages (Brown D., PNAS 94 : 13011-16, 1997). > > This is obviously an important issue, and certainly all zebrafish studies > dealing with stages past the first week of development?should refer to body > length in addition to or instead of nominal age. > Also, the community should agree about how to measure that length. In Figure 1 > and Table 2 of his chapter refered above, Tom Schilling provided two different > measurements : 'Full length' i.e. up to the tip of the caudal fin, and > 'standard length', ie up to the base of the caudal fin. > > I wasn't at the last Madison meeting but since there was a workshop there > about "working with post-embryonic stages", maybe this staging issue was > raised ? I would be interested to learn about this from participants to this > workshop.? > > Best wishes, > > Philippe? > > > ????????????????????????????????????????? > 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 > > > > > > Le 7 ao?t 08 ? 21:02, Oliver Braubach a ?crit : > >> Dear All. >> >> I have been working on aspects of postembryonic development in the zebrafish >> brain, and have been trying to determine a standard method for 'aging' >> zebrafish. As many of you may be aware, the development of individual fish >> varies greatly during the first month of development, which ultimately leads >> to very noticeable differences in the size of individual fish and their >> organs. >> >> Thus, age does not accurately characterize developmental stages of fish, and >> I have been trying to determine if there's another standard method to do so. >> Has anyone tried to correlate body length to fish age? Are there any >> citations for expected / optimal size of fish are a certain age? >> >> Any input regarding this problem would be greatly appreciated. >> Best, >> >> Oliver Braubach >> Department of Physiology and Biophysics >> Dalhousie University >> Halifax, NS B3H 1X5 Canada >> Tel: 902.494.6471 >> Fax: 902.494.1685 >> >> _______________________________________________ >> Zbrafish mailing list >> Zbrafish@net.bio.net >> http://www.bio.net/biomail/listinfo/zbrafish >> > > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > 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. > > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080808/f0d93638/attachment-0001.html From mena22787 from excite.com Tue Aug 12 08:15:12 2008 From: mena22787 from excite.com (Serena) Date: Tue Aug 12 11:34:35 2008 Subject: [Zbrafish] Re: Swarming zebrafish embryo parasites References: Message-ID: On Jul 31, 9:16 pm, RachelRay...@gmail.com wrote: > While performing maintenance on a clutch of 48 hour zebrafish embryos, > I came across one with a swarm of parasites within its chorion. All > the other embryos in the clutch were fine. I examined the embryo 5 > hours later and it was almost completely devoured. I assume that this > is a parasite that the mother passed on? Unfortunately, the cross was > albino stock and the individual fish was put back into a group tank > after breeding. What were those things and what action if any needs > to be taken? > > If anyone would like a picture or video of the embryo I’d be happy to > send one in hopes of identifying the parasite. Would you mind sending me a photo also? I'm rather curious to see what these embryos look like...Thanks! From dirkvandermierde from yahoo.com Tue Aug 12 04:03:10 2008 From: dirkvandermierde from yahoo.com (dirkvandermierde@yahoo.com) Date: Tue Aug 12 12:15:48 2008 Subject: [Zbrafish] FACS purification of zebrafish larvae liver cells Message-ID: <372d88ca-4066-4c0c-88e8-6677fcae6747@k7g2000hsd.googlegroups.com> Hi there! Does anybody have some experience in isolation of zebrafish larvae liver cells by FACS purification? Any input, weblinks or protocols would be welcome. Thx! From jocelyn.mcauley from gmail.com Tue Aug 12 13:21:37 2008 From: jocelyn.mcauley from gmail.com (Jocelyn) Date: Tue Aug 12 13:43:12 2008 Subject: [Zbrafish] purchased hanks for squeezing? Message-ID: Has anyone used Gibco's Hanks in an invitro fertilization protocol? I'm noticing that Gibco's formula includes glucose (to up the osmolarity?), where as the Zebrafish book recipe does not use glucose. Will this affect the viability of the sperm? I need to do a squeeze, and don't have the stock solution components on hand to make hanks, but instead have the Gibco premade 10x hanks (- bicarb, - phenol red). Any advice? From castrand from mail.nih.gov Wed Aug 13 08:54:07 2008 From: castrand from mail.nih.gov (Daniel Castranova) Date: Wed Aug 13 11:05:07 2008 Subject: [Zbrafish] purchased hanks for squeezing? In-Reply-To: References: Message-ID: <6BF1F380-065E-4E56-B5C7-5173AAC1E9D9@mail.nih.gov> Jocelyn, I have not used Gibco's hanks for IVF in zebrafish but many short term extenders used in other fish species include glucose. I don't think the slightly higher osmolarity will be a problem since high osmolarity sequesters sperm activation. I'd give it a shot. Good luck, 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 Aug 12, 2008, at 2:21 PM, Jocelyn wrote: > Has anyone used Gibco's Hanks in an invitro fertilization protocol? > > I'm noticing that Gibco's formula includes glucose (to up the > osmolarity?), where as the Zebrafish book recipe does not use glucose. > Will this affect the viability of the sperm? > > I need to do a squeeze, and don't have the stock solution components > on hand to make hanks, but instead have the Gibco premade 10x hanks (- > bicarb, - phenol red). > > Any advice? > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080813/132f9164/attachment.html From jocelyn.mcauley from gmail.com Thu Aug 14 12:07:06 2008 From: jocelyn.mcauley from gmail.com (Jocelyn) Date: Thu Aug 14 12:10:26 2008 Subject: [Zbrafish] Re: purchased hanks for squeezing? References: Message-ID: thanks for the feedback (1. just try it, 2. make up Ginsbergs) offered on and off the group. After deciding to give it a try with some other controls in place, I was faced with the hard reality of the females not yielding any eggs. Interestingly, these are fish that give eggs on a weekly basis in pair- wise set ups. I am all the wiser now and more appreciative of the squeeze-worthy selected AB fish. From jason.cockington from gmail.com Mon Aug 18 02:42:29 2008 From: jason.cockington from gmail.com (Leviathan) Date: Mon Aug 18 14:00:04 2008 Subject: [Zbrafish] 10th A&NZ Zf Workshop 2009 Message-ID: <092da702-d76e-4511-a761-5412238b4fab@p31g2000prf.googlegroups.com> Dear Colleagues, On behalf of the Adelaide Organising Committee, and our Major Sponsors ZebTec, NGED and the CMGD, it is with great pleasure that i invite you to register for the 10th Australia and New Zealand Zebrafish Workshop, hosted by the University of Adelaide Zebrafish Community, to be held at the Whalers Inn Resort, Victor Harbour, South Australia from the 2nd to the 4th of February, 2009. This meeting will be followed by a one day Zebrafish Husbandry Workshop to be held back at the University of Adelaide on the 5th of February, 2009. Registrations are now open, and will be accepted up until the 10th of October 2008. While registrations may be accepted beyond this date, please be aware that they will incur a late fee of $100 on top of the standard registration fee. For information on guest speakers, or about the meeting or its venue, see the official website: http://www.adelaide.edu.au/zebrafish09/ best fishes, jason From mena22787 from excite.com Mon Aug 18 15:19:48 2008 From: mena22787 from excite.com (Serena) Date: Mon Aug 18 18:01:47 2008 Subject: [Zbrafish] ze positive responses Message-ID: <16acb008-fd2f-4164-a4e0-f0d2a2eeacdc@v57g2000hse.googlegroups.com> Hi all! Our fish have stopped producing embryos so we have currently shifted gears. We now change out 10% of the water (we have a 10 gallon, 20 gallon, and a benchtop recirculation system) of each tank once daily with conditioned water. We have also been experimenting with different types of food, ranging from artemia to dried brine shrimp flakes to pellets. My question is how long should it take for fish to respond positively and resume embryo production if conditions are favorable? Thanks! Serena From shellyleibman from gmail.com Tue Aug 19 00:22:16 2008 From: shellyleibman from gmail.com (shellyleibman@gmail.com) Date: Tue Aug 19 11:05:34 2008 Subject: [Zbrafish] Bleaching embryos after In Situ Hybridization Message-ID: <1e712b41-5272-4245-9090-a271580c7e78@k7g2000hsd.googlegroups.com> Hello I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I wanted to ask if anyone knows how to bleach the pigment of 2dpf zebrafish embryos post whole mount In Situ Hybridization (it was not raised in PTU water). Many thanks in advance Shelly From nonolow from gmail.com Wed Aug 20 11:49:09 2008 From: nonolow from gmail.com (nonolow@gmail.com) Date: Wed Aug 20 11:52:14 2008 Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization References: Message-ID: <69b4622d-afa1-4d79-bcf1-d140589920dc@f36g2000hsa.googlegroups.com> bleaching is a routine approach in frog embryo whole mount in situ hybridization, i think you could find the protocal and recipes in the Frog Book (Early Development of Xenopus Laevis: A Laboratory Manual).... And be aware the time of bleaching, hydrogen peroxide could breakdown the staining of in situ hybridization. low On Aug 19, 1:22?am, shellyleib...@gmail.com wrote: > Hello > I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I > wanted to ask if anyone knows how to bleach the pigment of 2dpf > zebrafish embryos post whole mount In Situ Hybridization (it was not > raised in PTU water). > Many thanks in advance > > Shelly From nonolow from gmail.com Wed Aug 20 11:55:33 2008 From: nonolow from gmail.com (nonolow@gmail.com) Date: Wed Aug 20 11:56:53 2008 Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization References: Message-ID: <59eb3d60-d642-4604-87fc-52e9cd3dd91d@z72g2000hsb.googlegroups.com> bleaching is a routine approach in frog embryos, you could find some protocols and recipes in the book, Early Development of Xenopus Laevis: A Laboratory Manual, be aware the time of bleaching, hydrogen peroxide could breakdown the staining of in situ hybridization. good luck low On Aug 19, 1:22?am, shellyleib...@gmail.com wrote: > Hello > I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I > wanted to ask if anyone knows how to bleach the pigment of 2dpf > zebrafish embryos post whole mount In Situ Hybridization (it was not > raised in PTU water). > Many thanks in advance > > Shelly From rburdine from Princeton.EDU Wed Aug 20 12:31:16 2008 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Wed Aug 20 12:35:01 2008 Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization In-Reply-To: <59eb3d60-d642-4604-87fc-52e9cd3dd91d@z72g2000hsb.googlegroups.com> References: <59eb3d60-d642-4604-87fc-52e9cd3dd91d@z72g2000hsb.googlegroups.com> Message-ID: <04AF14C4EF3CF34682460FB1FDBA98C027D234@MBCLUSTER.pu.win.princeton.edu> Here is the blurb from our lab protocol which I think we cribbed from David Parichy's protocols? Part 4: Removing pigmentation in older embryos. Embryos start developing pigmentation at around 1dpf. Embryos can be de-pigmented using hydrogen peroxide. Embryos can be bleached right after PFA fixation (before MeOH storage) or upon rehydration after MeOH storage. If you do it on the first day of the in situ, simply add 3 additional 5' PBT washes. 1. Make a bleaching solution of 3% hydrogen peroxide and 1% KOH in dH20. You need 1ml of solution per tube of embryos to be bleached. Bleaching solution should be made fresh each time, since mixing the two reagents starts the chemical reaction! 100ul 30% Hydrogen peroxide (stored in large 4C) + 100ul 10% KOH (stored at RT) + 800ul Millipore water=1 mL 2. Aspirate PFA (or PBT) and add 1 mL of bleaching solution to each tube, and leave the tubes open to allow for gas escape. Typically 36-hpf embryos take 10-15 minutes, and 5-dpf embryos take ~45 min. Check the tubes occasionally in case the bubbles from the peroxide reaction push any embryos out the top of the tube. 3. When the embryos appear sufficiently bleached (monitor this closely), aspirate the bleaching solution and rinse 2-3 times in PBT, then continue to MeOH storage or the in situ protocol. You may want to remove the solution with a plastic transfer pipet instead of with the vacuum because the embryos will be floating on top and may get accidentally sucked up easily. After a couple PBT washes, the embryos will eventually return to the bottom of the tube. Note that the reaction times seem to depend upon the bleach, the temperature in the room etc. Watch a test batch closely and the minute you can no longer see black specs in the epitube, stop the reaction. Letting this go too long will chew up your embryos. In our hands bleaching is better than PTU as it doesn't affect development and can be done after fixation. Becky --------------------------------------------------- Rebecca D. Burdine, Ph.D. Assistant Professor Dept. of Molecular Biology Princeton University Washington Road Mof 433 Princeton, NJ 08544 Phone: (609) 258-7515 Fax: (609) 258-1343 Email: rburdine@princeton.edu Admin Assistant: Cathy Falk (609) 258-1604 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu > [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of > nonolow@gmail.com > Sent: Wednesday, August 20, 2008 12:56 PM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization > > bleaching is a routine approach in frog embryos, you could > find some protocols and recipes in the book, Early > Development of Xenopus > Laevis: A Laboratory Manual, > be aware the time of bleaching, hydrogen peroxide could > breakdown the staining of in situ hybridization. > good luck > > > low > > > > > > On Aug 19, 1:22?am, shellyleib...@gmail.com wrote: > > > > Hello > > I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv > University. I > > wanted to ask if anyone knows how to bleach the pigment of 2dpf > > zebrafish embryos post whole mount In Situ Hybridization > (it was not > > raised in PTU water). > > Many thanks in advance > > > > Shelly > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > From siberiamao from uchicago.edu Wed Aug 20 12:44:01 2008 From: siberiamao from uchicago.edu (Qiyan Mao) Date: Wed Aug 20 12:46:42 2008 Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization In-Reply-To: <200808201703.m7KH3TU28825@net.bio.net> Message-ID: Hi, I've recently bleached my 48 h embryos before ISH according to Thisse's protocol (Nature protocols (2008) 3(1):59) using 3% hydrogen peroxide in 0.5% KOH, and it worked fine for me. The whole bleaching takes around 20 minutes. My impression is that this bleaching step doesn't alter ISH staining. I also tried post-ISH bleaching using the same procedure, but for some reason the pigment doesn't go away as easily. Hope this could help. Best, Qiyan On 8/20/08 12:03 PM, zbrafish-request@oat.bio.indiana.edu wrote: > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Re: Bleaching embryos after In Situ Hybridization > (nonolow@gmail.com) > 2. Re: Bleaching embryos after In Situ Hybridization > (nonolow@gmail.com) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 20 Aug 2008 09:49:09 -0700 (PDT) > From: nonolow@gmail.com > Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization > To: bionet-organisms-zebrafish@moderators.isc.org > Message-ID: > <69b4622d-afa1-4d79-bcf1-d140589920dc@f36g2000hsa.googlegroups.com> > Content-Type: text/plain; charset=ISO-8859-1 > > bleaching is a routine approach in frog embryo whole mount in situ > hybridization, i think you could find the protocal and recipes in the > Frog Book (Early Development of Xenopus Laevis: A Laboratory > Manual).... > And be aware the time of bleaching, hydrogen peroxide could breakdown > the staining of in situ hybridization. > > > low > > > > On Aug 19, 1:22?am, shellyleib...@gmail.com wrote: >> Hello >> I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I >> wanted to ask if anyone knows how to bleach the pigment of 2dpf >> zebrafish embryos post whole mount In Situ Hybridization (it was not >> raised in PTU water). >> Many thanks in advance >> >> Shelly > > > > > ------------------------------ > > Message: 2 > Date: Wed, 20 Aug 2008 09:55:33 -0700 (PDT) > From: nonolow@gmail.com > Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization > To: bionet-organisms-zebrafish@moderators.isc.org > Message-ID: > <59eb3d60-d642-4604-87fc-52e9cd3dd91d@z72g2000hsb.googlegroups.com> > Content-Type: text/plain; charset=ISO-8859-1 > > bleaching is a routine approach in frog embryos, you could find some > protocols and recipes in the book, Early Development of Xenopus > Laevis: A Laboratory Manual, > be aware the time of bleaching, hydrogen peroxide could breakdown the > staining of in situ hybridization. > good luck > > > low > > > > > > On Aug 19, 1:22?am, shellyleib...@gmail.com wrote: > > >> Hello >> I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I >> wanted to ask if anyone knows how to bleach the pigment of 2dpf >> zebrafish embryos post whole mount In Situ Hybridization (it was not >> raised in PTU water). >> Many thanks in advance >> >> Shelly > > > > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 39, Issue 10 > **************************************** From csie2009 from shine2009.gnway.net Tue Aug 26 11:51:03 2008 From: csie2009 from shine2009.gnway.net (cmc2009) Date: Tue Aug 26 12:00:16 2008 Subject: [Zbrafish] CSIE 2009, Los Angeles, Submissions: September 30 Message-ID: <200808261657.m7QGvTU02876@net.bio.net> 2009 World Congress on Computer Science and Information Engineering (CSIE 2009) March 31 - April 2, 2009 Los Angeles/Anaheim, USA http://world-research-institutes.org/conferences/CSIE/2009 CALL FOR PAPERS & EXPO The Los Angeles/Anaheim area is known for its many renowned attractions, such as Disneyland, Universal Studios and the Hollywood Walk of Fame. Very few cities in the world offer as much entertainment, excitement and diversity as Los Angeles/Anaheim does. CSIE 2009 intends to be a global forum for researchers and engineers to present and discuss recent advances and new techniques in computer science and information engineering. CSIE 2009 consists of the following Technical Symposiums: * Communications & Mobile Computing Symposium * Computer Applications Symposium * Computer Design & VLSI Symposium * Data Mining & Data Engineering Symposium * Intelligent Systems Symposium * Multimedia & Signal Processing Symposium * Software Engineering Symposium CSIE 2009 conference proceedings will be published by the IEEE Computer Society and all papers in the proceedings will be included in EI Compendex, ISTP, and IEEE Xplore. In addition to research papers, CSIE 2009 also seeks exhibitions of modern products and equipment for computer science and information engineering. Important Dates: Paper/Abstract Submission Deadline: September 30, 2008 Review Notification: November 15, 2008 Final Papers and Author Registration Deadline: December 7, 2008 Organizing Committee: General Chair: Adrian Martin, World Research Institutes, USA Program Chair: Mark Burgin, University of California at Los Angeles, USA Symposium Chairs: Masud H Chowdhury, University of Illinois at Chicago, USA Chan H. Ham, University of Central Florida, USA Simone Ludwig, University of Saskatchewan, Canada Weilian Su, Naval Postgraduate School, USA Sumanth Yenduri, University of Southern Mississippi, USA Publicity Chair: Nitin Upadhyay, Birla Institute of Technology and Science (BITS), India David C. Wong, US Environmental Protection Agency, USA (Please forward to those who may be interested.) (To unsubscribe all WRI announcements, please email csie2009@world-research-institutes.org with subject "Unsubscribe ALL zbrafish@magpie.bio.indiana.edu". Thanks and apologies) (To unsubscribe CSIE announcements only, please email csie2009@world-research-institutes.org with subject "Unsubscribe CSIE zbrafish@magpie.bio.indiana.edu".) From hmcallis from Princeton.EDU Tue Aug 26 13:49:10 2008 From: hmcallis from Princeton.EDU (Heather S McAllister (hmcallis@Princeton.EDU)) Date: Tue Aug 26 14:19:53 2008 Subject: [Zbrafish] Re: Zbrafish Digest, Vol 20, Issue 15 In-Reply-To: <200701261700.l0QH0Dn26655@net.bio.net> References: <200701261700.l0QH0Dn26655@net.bio.net> Message-ID: some info on mycobacteria (Aka fish tuberculosis) Heather McAllister Research Specialist II Burdine Lab Department of Molecular Biology Princeton University Princeton, NJ 08544 hmcallis@princeton.edu 609-258-5782 ----- Original Message ----- From: zbrafish-request@oat.bio.indiana.edu Date: Friday, January 26, 2007 12:11 pm Subject: Zbrafish Digest, Vol 20, Issue 15 To: zbrafish@magpie.bio.indiana.edu > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Re: Mycobacteria (Jen Matthews) > > > ------------------------------------------------------------------- > --- > > Message: 1 > Date: Wed, 24 Jan 2007 17:51:26 -0800 > From: Jen Matthews > Subject: Re: [Zbrafish] Mycobacteria > To: Padnos.Beth@epamail.epa.gov > Cc: zbrafish@oat.bio.indiana.edu > Message-ID: <9A6DD50A-9FD7-48D2-B02E-60208F1FD927@zfin.org> > Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed > > Hi Beth, > I have just a few things to add to the discussion on mycobacteria. > > There seems to be a lot of hype and often over reaction to the > diagnosis mycobacteriosis in the zebrafish community. Atypical or > > nontuberculosis Mycobacterium spp. are ubiquitous in aquatic > environments, persisting largely in biofilms. I would argue that > if > you look hard enough, you will find it in all established > zebrafish > systems. Many Mycobacterium spp. are resistant to common chlorine > disinfection. That's why it is also commonly found in municipal > drinking water distribution systems and swimming pools. A number > of > these Mycobacterium spp. are documented pathogens of zebrafish, > however, they are opportunist pathogens causing chronic > infections. > Asymptomatic carriers of mycobacteria are common in zebrafish with > > the swimbladder and ovary being the most commonly infected organs. > > There are strains of zebrafish that are more susceptible to > mycobacteria infections. In our experience, the TU strain is ten > times more likely to be infected compared to other wild-type > strains. > There also appears to be some differences in virulence between the > > different species and strains of Mycobacterium. > > So with this background, what do you do? I am not a proponent of > nuking systems and starting over. You will most likely get the > same > bug back. All Mycobacterium spp. respond poorly to antibiotics, so > > this is also not a good option for fish facilities. Because of the > > opportunist nature of bacterial infections in fish, your first > focus > should be on husbandry and water quality. Stress lowers the immune > > competency of fish. This includes any type of suboptimal water > quality (e.g. elevated nitrogenous compounds - including nitrate, > pH > fluctuations) and other stressors (e.g. high stocking density, > over > use). The other focus should be on lowering the exposure dose. All > > pathogens have a dose response - the higher the exposure to > mycobacteria the greater the infection rate. To decrease the dose > of > bacteria make sure that your UV is properly sized and maintained, > that cleaning procedures are adequate, and do not keep moribund > (sick) or old fish. Eliminating Mycobacterium from zebrafish > systems > is not a realistic goal, it needs to be managed. > > Let me know if you have questions. We are also always happy to > give > feedback on your husbandry and water quality parameters. very > best, -Jen > > > > Jennifer L. Matthews, DVM, PhD > Zebrafish International Resource Center > Pathology and Health Services > 5274 University of Oregon > Eugene, OR 97403-5274 <(()>< > (541) 346-6028 ext. 14 <(()>< > Fax (541) 346-6151 > jmatthews@zfin.org > > > On Jan 23, 2007, at 8:07 AM, Padnos.Beth@epamail.epa.gov wrote: > > > I work in a facility that is trying to establish a small Medaka and > > Zebra fish colony. > > Recently we discovered Mycobacteria in our Medaka which are on > the > > same > > rack as our Zebrafish. One of the male Zebrafish began swimming > a bit > > oddly and his abdomen became red as if bleeding internally or > inflamed> organs. He is has since been set to the pathologist for > screening, but > > is assumed to have Mycobacteria as well. > > The facility Veterinarian and I have both done literature > searches but > > find not treatment. Has anyone successfully treated or eliminated > > Mycobacteria from their fish and or system? > > Help is greatly appreciated from an inexperienced fish researcher. > > Beth Padnos > > > > _______________________________________________ > > Zbrafish mailing list > > Zbrafish@net.bio.net > > http://www.bio.net/biomail/listinfo/zbrafish > > > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 20, Issue 15 > **************************************** > From bobbi from aquaneering.com Thu Aug 28 11:52:25 2008 From: bobbi from aquaneering.com (Bobbi Baur) Date: Thu Aug 28 15:26:07 2008 Subject: [Zbrafish] Zebrafish food Message-ID: <0D939144273ADA4D8B38EC00A764E319549089@BE05.exg4.exghost.com> Hi Sarah, Aquaneering offers a Scientific Hatcheries 3-pigment diet developed specifically for Zebrafish. Please contact us off-list if you would like a sample. Bobbi M. Baur Sales Manager/Aquaneering 7960 Stromesa Ct. San Diego, CA 92126 858-578-2028 (ph) 858-689-9326 (fx) www.aquaneering.com EMAIL CONFIDENTIALITY NOTICE: The information contained in this E-Mail transmission, including any attachments, is confidential, proprietary or privileged and may be subject to protection under the law. This message is intended for the sole use of the individual or entity to which it is addressed. If you are not the intended recipient, you are notified that any use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalties. If you received this transmission in error, please contact the sender by replying to this E-mail and delete this email immediately. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080828/79676abd/attachment.html From moconnel from tcnj.edu Fri Aug 29 14:37:09 2008 From: moconnel from tcnj.edu (moconnel) Date: Fri Aug 29 15:20:15 2008 Subject: [Zbrafish] tenure track positions in biology Message-ID: <48B84FE5.9020708@tcnj.edu> The Department of Biology at The College of New Jersey will be conducting three searches for tenure-track faculty, as outlined below: THREE TENURE-TRACK BIOLOGY FACULTY POSITIONS Organismal Biology, Genetics, and Biology Education/Pedagogy The Department of Biology at The College of New Jersey (TCNJ) invites outstanding applicants for three tenure-track faculty positions, starting August 2009. Teaching and research are mutually supportive activities at TCNJ. Candidates should be strongly committed to the teacher-scholar model in a primarily undergraduate, residential institution and to maintaining both high quality teaching and an active and productive research program involving highly motivated undergraduates. Faculty members also serve as academic advisors and have service responsibilities within the College. A research laboratory and competitive start-up funds will be provided. The Biology Department is housed in a modern biology building that offers state-of-the-art teaching and research facilities and instrumentation. For further information about our program, please visit: http://www.tcnj.edu/~biology/ . We seek broadly trained candidates who also have potential to contribute collaboratively to interdisciplinary curricular and scholarly efforts within the School of Science and at the College. In addition to the courses listed below, teaching responsibilities may include rotation through either majors and/or non-majors introductory courses. 1) *Organismal Biology *(Assistant Professor) – to teach a junior/senior-level course that fulfills the department’s organismal-level biology requirement, an upper-level course in area of specialty, and one of our core courses. Research in any area of animal organismal biology will be considered. 2) *Genetics *(Assistant Professor) –* *to teach our core course in genetics and an upper-level course(s) in area of specialty. Research in any area of genetics will be considered. 3) *Biology Education/Pedagogy *(Assistant or Associate Professor) – to teach an interdisciplinary science course to elementary education majors, a methods course to science majors preparing for secondary teacher certification, and potentially a biology course in area of specialty; conduct research in the area of biology education/pedagogy; and provide leadership in emerging science education initiatives and funding opportunities. TCNJ is a highly selective, public institution that has earned national recognition for its commitment to excellence. TCNJ emphasizes the residential experience for its 5,900 undergraduate students, who benefit from a 12-to-1 student-to-faculty ratio and an average class size of only 20 students. TCNJ is one of only five public institutions included on /Barron//’//s Profiles of American Colleges/ list of the 75 “most competitive” colleges and universities in the United States, and of those five, TCNJ is the only primarily undergraduate institution included. TCNJ’s campus encompasses 289 beautiful tree-lined acres in suburban Ewing, NJ, which is located between Trenton and Princeton in the historic Delaware Valley, and is convenient to Philadelphia, New York, the Jersey shore, and the Pocono Mountains. Candidates should have a Ph.D. (or equivalent in education, content pedagogy, or a related field for the Education position) and post-doctoral experience is preferred (or appropriate experience at the P-20 level for Education position); however, meritorious ABD candidates will be considered. To apply, send a letter of application; current curriculum vitae; statement of teaching philosophy; statement of research interests and goals; representative publications; all graduate and undergraduate transcripts; and three letters of recommendation to: Faculty Search (indicate position), Department of Biology, The College of New Jersey, P.O. Box 7718, Ewing, NJ 08628. All materials must be received as hard copies. Review of applications will begin September 15, 2008 and will continue until the positions are filled. To enrich education through diversity, TCNJ is an Affirmative Action /Equal Opportunity Employer. Women and members of underrepresented groups are encouraged to apply. -------------- next part -------------- A non-text attachment was scrubbed... Name: moconnel.vcf Type: text/x-vcard Size: 274 bytes Desc: not available Url : http://www.bio.net/bionet/mm/zbrafish/attachments/20080829/06a35195/moconnel.bin From Padnos.Beth from epamail.epa.gov Fri Aug 29 15:11:09 2008 From: Padnos.Beth from epamail.epa.gov (Padnos.Beth@epamail.epa.gov) Date: Fri Aug 29 15:31:50 2008 Subject: [Zbrafish] tank cleaning In-Reply-To: <200808291703.m7TH37U19621@net.bio.net> Message-ID: 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.