lechristophe |
Dear imaging specialists,
I'm wondering if tranporting cell cultures (low-density rat hippocampal neurons, to be more specific) would be feasible from my lab to an imaging facility that is between half an hour and an hour away. I'm surprized how little information I could get on the web. Do such thing as a portable incubator exist ? Do any of you have a low-tech / low-price advice to give ? I'd be glad to hear your suggestions. Thanks a lot, Christophe Leterrier Ionic channels neurobiology Lab UMR641 - Marseille, France |
Michael Schell |
I've transported hippocampal neurons about half an hour away, but I never tried longer. To maximize your chances of success:
1. The best plan is to transport them between 3 and 7 days after plating, before they become synaptically mature. If you can bring them to the imaging location during that time and allow them to recover in an incubator for a day or longer, that's the best solution. 2. Add Hepes pH 7.2 to a final concentration of 10-20 mM to each dish to be transported. Then, seal the culture dishes with parafilm and place them stacked in a styrofoam box for transport. Drive carefully. 3. Your neurons will survive best if you can raise the MgCl concentration in the medium up to 10 mM just before transport. This will greatly reduce deadly calcium influx. However, this maneuver might not be compatible with your experiment, since cultured hippocampal neurons require conditioned medium to survive (you cannot just change out of the Hi Mg medium into fresh medium once you get to the imaging destination; having extra conditioned medium available at the destination might circumvent this problem). Good luck, Mike Michael J. Schell, Ph.D., CIV, USUHS Assist. Professor Dept. of Pharmacology Uniformed Services University 4301 Jones Bridge Rd. Bethesda, MD 20814-3220 tel: (301) 295-3249 [hidden email] >>> Christophe Leterrier <[hidden email]> 02/04/09 5:48 PM >>> Dear imaging specialists, I'm wondering if tranporting cell cultures (low-density rat hippocampal neurons, to be more specific) would be feasible from my lab to an imaging facility that is between half an hour and an hour away. I'm surprized how little information I could get on the web. Do such thing as a portable incubator exist ? Do any of you have a low-tech / low-price advice to give ? I'd be glad to hear your suggestions. Thanks a lot, Christophe Leterrier Ionic channels neurobiology Lab UMR641 - Marseille, France |
Pablo German |
In reply to this post by lechristophe
Hi Christophe,
I've been trying to transport HEK293 cell cultures but they were not making it in good condition. I've been advised to use a carbonated buffer such as Hanks' solution (HBSS). I haven't tried it yet though. I'm as interested as you are to see what solutions are possible.
Regards,
Pablo
2009/2/5 Christophe Leterrier <[hidden email]> Dear imaging specialists, -- Pablo German Plant and Food Research Private Bag 92169 Auckland Mail Centre Auckland 1142 New Zealand DDI: (09) 925-7107 Mobile: 0210459406 |
Jenni Poole |
In reply to this post by lechristophe
Hi Christophe,
There are portable incubators available that would work for the type of transport you are searching for. Minitube of America offers such a device. With the following link you can find additional information. http://www.minitube.com/catalog_items.asp?ProdGrp_ID=173&Cat_ID=4 This device would definitely work for your short transport. It even has a cord that can connect in a car or once charged it can be shipped UPS/FedEx Next Day Air to another location. The portable incubator was originally designed for embryo transport. Let me know if you would like additional information. Thanks, Jenni Poole Sales Penetrating Innovations LLC ibidi LLC Minitube of America 419 Venture Ct. Verona, WI 53593 Phone (608) 845-3270 Fax (608) 845-3271 [hidden email] www.pi-ivf.com www.ibidi.com www.minitube.com -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christophe Leterrier Sent: Wednesday, February 04, 2009 4:37 PM To: [hidden email] Subject: Cell culture transport (one hour drive) possible ? Dear imaging specialists, I'm wondering if tranporting cell cultures (low-density rat hippocampal neurons, to be more specific) would be feasible from my lab to an imaging facility that is between half an hour and an hour away. I'm surprized how little information I could get on the web. Do such thing as a portable incubator exist ? Do any of you have a low-tech / low-price advice to give ? I'd be glad to hear your suggestions. Thanks a lot, Christophe Leterrier Ionic channels neurobiology Lab UMR641 - Marseille, France |
Phillips, Thomas E. |
In reply to this post by Michael Schell
I have never transported hippocampal neurons but have shipped growing
cells internationally on many occasions so we are taking 1-3 days in transit. You want to avoid sloshing that would either temporarily dry them out or physically beat on them like waves hitting the shore. I fill my flasks to the very tip top and seal tightly. This won't work for cells on coverslips or multiwells but should if they are in flasks. I agree Hepes buffer is needed. Thomas E. Phillips, Ph.D Professor of Biological Sciences Chair, MU Faculty Council Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400 573-882-4712 (office) 573-882-0123 (fax) [hidden email] http://www.biology.missouri.edu/faculty/phillips.html http://www.biotech.missouri.edu/mcc/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Michael Schell Sent: Wednesday, February 04, 2009 5:01 PM To: [hidden email] Subject: Re: Cell culture transport (one hour drive) possible ? I've transported hippocampal neurons about half an hour away, but I never tried longer. To maximize your chances of success: 1. The best plan is to transport them between 3 and 7 days after plating, before they become synaptically mature. If you can bring them to the imaging location during that time and allow them to recover in an incubator for a day or longer, that's the best solution. 2. Add Hepes pH 7.2 to a final concentration of 10-20 mM to each dish to be transported. Then, seal the culture dishes with parafilm and place them stacked in a styrofoam box for transport. Drive carefully. 3. Your neurons will survive best if you can raise the MgCl concentration in the medium up to 10 mM just before transport. This will greatly reduce deadly calcium influx. However, this maneuver might not be compatible with your experiment, since cultured hippocampal neurons require conditioned medium to survive (you cannot just change out of the Hi Mg medium into fresh medium once you get to the imaging destination; having extra conditioned medium available at the destination might circumvent this problem). Good luck, Mike Michael J. Schell, Ph.D., CIV, USUHS Assist. Professor Dept. of Pharmacology Uniformed Services University 4301 Jones Bridge Rd. Bethesda, MD 20814-3220 tel: (301) 295-3249 [hidden email] >>> Christophe Leterrier <[hidden email]> 02/04/09 5:48 PM >>> Dear imaging specialists, I'm wondering if tranporting cell cultures (low-density rat hippocampal neurons, to be more specific) would be feasible from my lab to an imaging facility that is between half an hour and an hour away. I'm surprized how little information I could get on the web. Do such thing as a portable incubator exist ? Do any of you have a low-tech / low-price advice to give ? I'd be glad to hear your suggestions. Thanks a lot, Christophe Leterrier Ionic channels neurobiology Lab UMR641 - Marseille, France |
Sylvie Le Guyader-2 |
In reply to this post by lechristophe
Hi Christophe
I think that's no problem. You need to seed them in a flask with air tight lid (not with filter) and fill the flask with fresh and pre-warmed/pre-CO2 balanced medium. Keep it in the incubator until you leave then use anything to prevent the temperature from falling too fast (eg a good outdoor jacket). Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader Dept of Biosciences and Nutrition Karolinska Institutet Novum 14157 Huddinge Sweden +46 (0)8 608 9240 > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Christophe > Leterrier > Sent: 04 February 2009 23:37 > To: [hidden email] > Subject: Cell culture transport (one hour drive) possible ? > > Dear imaging specialists, > > I'm wondering if tranporting cell cultures (low-density rat > hippocampal neurons, to be more specific) would be feasible from my > lab to an imaging facility that is between half an hour and an hour > away. I'm surprized how little information I could get on the web. Do > such thing as a portable incubator exist ? Do any of you have a > low-tech / low-price advice to give ? I'd be glad to hear your > suggestions. > > Thanks a lot, > > Christophe Leterrier > Ionic channels neurobiology Lab > UMR641 - Marseille, France |
RICHARD BURRY |
In reply to this post by Michael Schell
Do not for get mechanical damage! Media sloshing back and for will tear the cells off the substrate. It is best to fill the chamber completely (no air bubbles) with media before transport. This extra media also serves to keep the temperature more stable.
Also it is best to transport the culture a day before use so the cells can get settled down again. Dick Richard W. Burry, Ph.D. Department of Neuroscience, College of Medicine Campus Microscopy and Imaging Facility, Director The Ohio State University Associate Editor, Journal of Histochemistry and Cytochemistry 277 Biomedical Research Tower 460 West Twelfth Avenue Columbus, Ohio 43210 Voice 614.292.2814 Cell 614.638.3345 Fax 614.247.8849 ----- Original Message ----- From: Michael Schell <[hidden email]> Date: Wednesday, February 4, 2009 6:01 pm Subject: Re: Cell culture transport (one hour drive) possible ? To: [hidden email] > I've transported hippocampal neurons about half an hour away, > but I never tried longer. To maximize your chances of success: > > 1. The best plan is to transport them between 3 and 7 days > after plating, before they become synaptically mature. If > you can bring them to the imaging location during that time and > allow them to recover in an incubator for a day or longer, > that's the best solution. > > 2. Add Hepes pH 7.2 to a final concentration of 10-20 mM > to each dish to be transported. Then, seal the culture > dishes with parafilm and place them stacked in a styrofoam box > for transport. Drive carefully. > > 3. Your neurons will survive best if you can raise the > MgCl concentration in the medium up to 10 mM just before > transport. This will greatly reduce deadly calcium > influx. However, this maneuver might not be compatible > with your experiment, since cultured hippocampal neurons require > conditioned medium to survive (you cannot just change out of the > Hi Mg medium into fresh medium once you get to the imaging > destination; having extra conditioned medium available at the > destination might circumvent this problem). > > Good luck, > Mike > > > > > Michael J. Schell, Ph.D., CIV, USUHS > Assist. Professor > Dept. of Pharmacology > Uniformed Services University > 4301 Jones Bridge Rd. > Bethesda, MD 20814-3220 > tel: (301) 295-3249 > [hidden email] > >>> Christophe Leterrier <[hidden email]> > 02/04/09 5:48 PM >>> > Dear imaging specialists, > > I'm wondering if tranporting cell cultures (low-density rat > hippocampal neurons, to be more specific) would be feasible from my > lab to an imaging facility that is between half an hour and an hour > away. I'm surprized how little information I could get on the > web. Do > such thing as a portable incubator exist ? Do any of you have a > low-tech / low-price advice to give ? I'd be glad to hear your > suggestions. > > Thanks a lot, > > Christophe Leterrier > Ionic channels neurobiology Lab > UMR641 - Marseille, France > > > -- > BEGIN-ANTISPAM-VOTING-LINKS > ------------------------------------------------------ > > Teach CanIt if this mail (ID 797471698) is spam: > Spam: > https://antispam.osu.edu/b.php?c=s&i=797471698&m=17bf03a7c26bNot > spam: https://antispam.osu.edu/b.php?c=n&i=797471698&m=17bf03a7c26b > Forget vote: > https://antispam.osu.edu/b.php?c=f&i=797471698&m=17bf03a7c26b---- > -------------------------------------------------- > END-ANTISPAM-VOTING-LINKS > |
Carol Heckman |
In reply to this post by Pablo German
Hi Christophe-
One of my students adapted a little cooler meant for carrying drinks to this purpose. He made a little holder, just big enough for the 60-mm petri dish, and attached it to a plate that matched the dimensions of the inner lining. Then he filled the cooler with sand, put it in a 60 degree oven (Centigrade) and brought it out just before loading the culture dishes for transport. By carefully adjusting the composition of the plate, he could get the temperature on the plate to exactly 36-38 degrees, which the cells tolerated fine. Our cells did not like to cool down, but they didn't mind the pH going high. He just transported them in the dishes and medium they were already sitting in. He allowed them 2 hours in the incubator on the other end, to equilibrate the pH, and they were ready to go. They would stand an hour of transport in the carrier. carol Center for Microscopy & Microanalysis Bowling Green State University ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Pablo German [[hidden email]] Sent: Wednesday, February 04, 2009 6:00 PM To: [hidden email] Subject: Re: Cell culture transport (one hour drive) possible ? Hi Christophe, I've been trying to transport HEK293 cell cultures but they were not making it in good condition. I've been advised to use a carbonated buffer such as Hanks' solution (HBSS). I haven't tried it yet though. I'm as interested as you are to see what solutions are possible. Regards, Pablo 2009/2/5 Christophe Leterrier <[hidden email]<mailto:[hidden email]>> Dear imaging specialists, I'm wondering if tranporting cell cultures (low-density rat hippocampal neurons, to be more specific) would be feasible from my lab to an imaging facility that is between half an hour and an hour away. I'm surprized how little information I could get on the web. Do such thing as a portable incubator exist ? Do any of you have a low-tech / low-price advice to give ? I'd be glad to hear your suggestions. Thanks a lot, Christophe Leterrier Ionic channels neurobiology Lab UMR641 - Marseille, France -- Pablo German Plant and Food Research Private Bag 92169 Auckland Mail Centre Auckland 1142 New Zealand DDI: (09) 925-7107 Mobile: 0210459406 |
Patrick Van Oostveldt |
In reply to this post by RICHARD BURRY
Dear,
We tested these conditions and the effect of vibration on different cell cultures. The results of these experiments were published in acta astronautica, because it was a simulation of the launching conditions of the space shuttle. Baert P., Van Cleynenbreugel T., Vandesompele J., De Schynkel S., Vander Sloten J., Van Oostveldt P.: The potential (radio-)biological impact of launch vibration. Acta Astronautica. 58, 456-463 (2006) If you like to have a reprint, please send a personal mail. Patrick Van Oostveldt Quoting RICHARD BURRY <[hidden email]>: > Do not for get mechanical damage! Media sloshing back and for will > tear the cells off the substrate. It is best to fill the chamber > completely (no air bubbles) with media before transport. This extra > media also serves to keep the temperature more stable. > Also it is best to transport the culture a day before use so the > cells can get settled down again. > > Dick > > Richard W. Burry, Ph.D. > Department of Neuroscience, College of Medicine > Campus Microscopy and Imaging Facility, Director > The Ohio State University > Associate Editor, Journal of Histochemistry and Cytochemistry > 277 Biomedical Research Tower > 460 West Twelfth Avenue > Columbus, Ohio 43210 > Voice 614.292.2814 Cell 614.638.3345 Fax 614.247.8849 > > ----- Original Message ----- > From: Michael Schell <[hidden email]> > Date: Wednesday, February 4, 2009 6:01 pm > Subject: Re: Cell culture transport (one hour drive) possible ? > To: [hidden email] > >> I've transported hippocampal neurons about half an hour away, >> but I never tried longer. To maximize your chances of success: >> >> 1. The best plan is to transport them between 3 and 7 days >> after plating, before they become synaptically mature. If >> you can bring them to the imaging location during that time and >> allow them to recover in an incubator for a day or longer, >> that's the best solution. >> >> 2. Add Hepes pH 7.2 to a final concentration of 10-20 mM >> to each dish to be transported. Then, seal the culture >> dishes with parafilm and place them stacked in a styrofoam box >> for transport. Drive carefully. >> >> 3. Your neurons will survive best if you can raise the >> MgCl concentration in the medium up to 10 mM just before >> transport. This will greatly reduce deadly calcium >> influx. However, this maneuver might not be compatible >> with your experiment, since cultured hippocampal neurons require >> conditioned medium to survive (you cannot just change out of the >> Hi Mg medium into fresh medium once you get to the imaging >> destination; having extra conditioned medium available at the >> destination might circumvent this problem). >> >> Good luck, >> Mike >> >> >> >> >> Michael J. Schell, Ph.D., CIV, USUHS >> Assist. Professor >> Dept. of Pharmacology >> Uniformed Services University >> 4301 Jones Bridge Rd. >> Bethesda, MD 20814-3220 >> tel: (301) 295-3249 >> [hidden email] >> >>> Christophe Leterrier <[hidden email]> >> 02/04/09 5:48 PM >>> >> Dear imaging specialists, >> >> I'm wondering if tranporting cell cultures (low-density rat >> hippocampal neurons, to be more specific) would be feasible from my >> lab to an imaging facility that is between half an hour and an hour >> away. I'm surprized how little information I could get on the >> web. Do >> such thing as a portable incubator exist ? Do any of you have a >> low-tech / low-price advice to give ? I'd be glad to hear your >> suggestions. >> >> Thanks a lot, >> >> Christophe Leterrier >> Ionic channels neurobiology Lab >> UMR641 - Marseille, France >> >> >> -- >> BEGIN-ANTISPAM-VOTING-LINKS >> ------------------------------------------------------ >> >> Teach CanIt if this mail (ID 797471698) is spam: >> Spam: >> https://antispam.osu.edu/b.php?c=s&i=797471698&m=17bf03a7c26bNot >> spam: > https://antispam.osu.edu/b.php?c=n&i=797471698&m=17bf03a7c26b >> Forget vote: >> https://antispam.osu.edu/b.php?c=f&i=797471698&m=17bf03a7c26b---- >> -------------------------------------------------- >> END-ANTISPAM-VOTING-LINKS >> > > -- Dep. Moleculaire Biotechnologie Coupure links 653 B 9000 GENT tel 09 264 5969 fax 09 264 6219 |
Lesley Weston |
In reply to this post by lechristophe
A visiting scientist once kept a fibroblast culture alive during a
long trip from our lab to his home country. He completely filled with medium two flasks containing confluent attached cultures of cells , then carried them in the inside pockets of his jacket, which he wore the whole time. It worked, the cells were fine, but I don't know what he did about airport security. Lesley Weston. On 4-Feb-09, at 2:37 PM, Christophe Leterrier wrote: > Dear imaging specialists, > > I'm wondering if tranporting cell cultures (low-density rat > hippocampal neurons, to be more specific) would be feasible from my > lab to an imaging facility that is between half an hour and an hour > away. I'm surprized how little information I could get on the web. Do > such thing as a portable incubator exist ? Do any of you have a > low-tech / low-price advice to give ? I'd be glad to hear your > suggestions. > > Thanks a lot, > > Christophe Leterrier > Ionic channels neurobiology Lab > UMR641 - Marseille, France |
Jerry Sedgewick-2 |
In reply to this post by Carol Heckman
The University of South Carolina School of Medicine will again be
hosting a Basic Confocal Microscopy Workshop. This year’s workshop will be from June 15-19, 2009 and will include a series of lectures on the theory and applications of confocal microscopy, specimen preparation, processing confocal images in Photoshop, and 3D reconstructions using AMIRA. Students will be able to process triple labeled samples (cell cultures and sections) on site or bring their own samples to the workshop. Faculty will include Drs. Jay Jerome (Vanderbilt), Ralph Albrecht (Univ Wisconsin-Madison), John Mackenzie (North Carolina State Univ), Tom TrusK (Medical Univ South Carolina) and myself. Instruments and applications experts from Leica, Nikon, Olympus, Perkin Elmer, Photometrics, and Zeiss are expected to be available for hands on training and imaging of samples. Please register early as the last 4 workshops have been overbooked. Deadline for registration is June 1 as long as slots are available. For further information and registration go to: http://dba.med.sc.edu/irf/price/irf/irf.htm or contact Anna McNeal ([hidden email] <mailto:[hidden email]>) Bob Bob Price Research Professor Dept Cell Biol and Anat USC School of Medicine 6439 Garner's Ferry Road Columbia, SC 29208 Tel: 803-733-3392 Admin Tel: 803-253-5822 Fax: 803-733-3212 --- Get FREE High Speed Internet from USFamily.Net! -- http://www.usfamily.net/mkt-freepromo.html --- |
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