Microscope Stage Incubators
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Charles Matthew Blaha |
Microscope Stage Incubators
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hey everyone..this is my first time using this server. I am a new research assistant working in the department of bioengineering at Stanford University and I have a few questions regarding Microscope Stage Incubators. My group plans to study live corneal epithelial cell migration and proliferation (and possibly other cell characteristics) on transparent hydrogels over a period of 1 to 4 days. We plan to use 10 ro 20x objectives so it will be a long working distance. As an undergrad, I performed live cell imaging with a microscope stage incubator on a leica microscope, so I have some experiene in the area. I am in charge of determining which microscope stage incubator is best for our application. We have a Nikon TE300 with an Applied Scientific Instruments MS-2000 stage. There appears to be two different microscope stage incubation systems: hood incubators and stage incubators. I have spoken to a variety of companies with conflicting recommendations, although I am leaning towards a Stage incubator from OKO-labs. Any suggestions if a hood or a stage incubator is better? Has anyone had success or been disapointed with certain companies hoods or stage incubators? I would greatly appreciate any help. Sincerely, Charles Blaha M.S. Department of Bioengineering Stanford University |
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Charles, We do a lot of live cell imaging (also for longer period ~72 hours). Our experience showed that the box type incubators work better for this type of experiments. They clearly need a longer equilibration time (you need to switch them on 1-2 hours before your experiments), but afterwards offer superior stability. If you have a "decent" microscope, the focus drift over 24 hours is around the 0.5 micron. I need to mention, that we have air-conditioning in our rooms which is also advantageous in order to stabilize the conditions. (Simply speaking - it is also essential....) Of course if for any reason you need to change the temperature in a fast manner - you shouldn't take these. We have three different incubators in our facility (Solent Scientific, the "Ludin-box and cube" from LIS, and incubators from the EMBL Heidelberg). All of these work fine in our hand, but there are considerable price differences (especially if you need also CO2 control). The Oko-lab system is quite similar to these so probabyl it will also do the job. If you have auto-focus (hardware) on your system, then you can choose almost anything - we have done experiments even with a slightly modified hair-dryer. I have no commercial interest in any of the above mentioned products. Cheers Gabor -- Gabor Csucs Light Microscopy Centre, ETH Zurich Schafmattstrasse 18, HPM F16 CH-8093, Zurich, Switzerland Web: www.lmc.ethz.ch Phone: +41 44 633 6221 Fax: +41 44 632 1298 e-mail: [hidden email] |
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George McNamara |
Re: Microscope Stage Incubators
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In reply to this post by Charles Matthew Blaha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Charles, Check out the Precautionary Note section of the web page http://www.bioptechs.com/Products/Rebuild/rebuilding_service.html At 02:29 PM 11/13/2007, you wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hey everyone..this is my first time using this server. I am a new research >assistant working in the department of bioengineering at Stanford >University and I have a few questions regarding Microscope Stage >Incubators. > >My group plans to study live corneal epithelial cell migration and >proliferation (and possibly other cell characteristics) on transparent >hydrogels over a period of 1 to 4 days. We plan to use 10 ro 20x >objectives so it will be a long working distance. As an undergrad, I >performed live cell imaging with a microscope stage incubator on a leica >microscope, so I have some experiene in the area. I am in charge of >determining which microscope stage incubator is best for our application. >We have a Nikon TE300 with an Applied Scientific Instruments MS-2000 >stage. > >There appears to be two different microscope stage incubation systems: hood >incubators and stage incubators. I have spoken to a variety of companies >with conflicting recommendations, although I am leaning towards a Stage >incubator from OKO-labs. Any suggestions if a hood or a stage incubator is >better? Has anyone had success or been disapointed with certain companies >hoods or stage incubators? > >I would greatly appreciate any help. > >Sincerely, > >Charles Blaha M.S. >Department of Bioengineering >Stanford University George McNamara, Ph.D. University of Miami, Miller School of Medicine Image Core Miami, FL 33010 [hidden email] [hidden email] 305-243-8436 office http://home.earthlink.net/~pubspectra/ http://home.earthlink.net/~geomcnamara/ http://www.sylvester.org/health_pro/shared_resources/index.asp (see Analytical Imaging Core Facility) |
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal George McNamara wrote: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hi Charles, > > Check out the Precautionary Note section of the web page > http://www.bioptechs.com/Products/Rebuild/rebuilding_service.html > Note that the Ludin scope incubator system restricts the humidified and CO2 rich atmosphere to a small volume on the stage. --aryeh -- Aryeh Weiss School of Engineering Bar Ilan University Ramat Gan 52900 Israel Ph: 972-3-5317638 FAX: 972-3-7384050 |
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Michael Weber-4 |
Re: Microscope Stage Incubators
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In reply to this post by Charles Matthew Blaha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Charles, with long working distance objectives you can in theory go for a stage incubator. However, if your room temperature is not absolutely stable all the time, then you will soon run into stage drift problems. For such long experiments I would definitely go for a cage incubator (already mentioned: LIS, OKOlab). If you heat it up ca. half a day before, the whole microscope is at temperature and focus drift should be no problem anymore. If you go for a stage incubator in a room with unstable temperature, then I am quite sure that you need some kind of autofocus. Good software solutions can also do the job - depends on the sample, i.e. phase contrast autofocus or Z-scan focus which looks for the coverslip reflection. As already mentioned, the main advantages of stage incubators are speed (especially if you want to change between temperatures) and mobility. Chambers based on water-circulation are nice and stable, i.e. the OKOlab solution. cheers, Michael Charles Blaha wrote: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hey everyone..this is my first time using this server. I am a new research > assistant working in the department of bioengineering at Stanford > University and I have a few questions regarding Microscope Stage > Incubators. > > My group plans to study live corneal epithelial cell migration and > proliferation (and possibly other cell characteristics) on transparent > hydrogels over a period of 1 to 4 days. We plan to use 10 ro 20x > objectives so it will be a long working distance. As an undergrad, I > performed live cell imaging with a microscope stage incubator on a leica > microscope, so I have some experiene in the area. I am in charge of > determining which microscope stage incubator is best for our application. > We have a Nikon TE300 with an Applied Scientific Instruments MS-2000 > stage. > > There appears to be two different microscope stage incubation systems: hood > incubators and stage incubators. I have spoken to a variety of companies > with conflicting recommendations, although I am leaning towards a Stage > incubator from OKO-labs. Any suggestions if a hood or a stage incubator is > better? Has anyone had success or been disapointed with certain companies > hoods or stage incubators? > > I would greatly appreciate any help. > > Sincerely, > > Charles Blaha M.S. > Department of Bioengineering > Stanford University |
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In reply to this post by Charles Matthew Blaha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
dear Charles,
We have a live cell system. we use a stage incubator. I think it is ok for long term(48h) imaging. The cell can live well. We have ever run across three problems in our hand. the first is the Z-shift,although we have air condotion. so we bought one autofocus controller from ludl company, and it is really good.Second problem is that the cell is easy to die when doing fluorescence imaging, especially under UV light. we had better reduce the exciting light as dim as we can. so you had better have a cool CCD with high sensitive.the last problem is the dish. because we need to observe some tiny structure in cell such as tubelin.so we need 60 oil objectivr, but the working distance is small and the bottom of common dish is thick. We bought petri-dish to solve the problem.this kind of dish is also better for fluorescence imaging. Hope these are helpful for you. best regards zhan cheng > Date: Tue, 13 Nov 2007 14:29:47 -0500 > From: [hidden email] > Subject: Microscope Stage Incubators > To: [hidden email] > > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hey everyone..this is my first time using this server. I am a new research > assistant working in the department of bioengineering at Stanford > University and I have a few questions regarding Microscope Stage > Incubators. > > My group plans to study live corneal epithelial cell migration and > proliferation (and possibly other cell characteristics) on transparent > hydrogels over a period of 1 to 4 days. We plan to use 10 ro 20x > objectives so it will be a long working distance. As an undergrad, I > performed live cell imaging with a microscope stage incubator on a leica > microscope, so I have some experiene in the area. I am in charge of > determining which microscope stage incubator is best for our application. > We have a Nikon TE300 with an Applied Scientific Instruments MS-2000 > stage. > > There appears to be two different microscope stage incubation systems: hood > incubators and stage incubators. I have spoken to a variety of companies > with conflicting recommendations, although I am leaning towards a Stage > incubator from OKO-labs. Any suggestions if a hood or a stage incubator is > better? Has anyone had success or been disapointed with certain companies > hoods or stage incubators? > > I would greatly appreciate any help. > > Sincerely, > > Charles Blaha M.S. > Department of Bioengineering > Stanford University 使用新一代 Hotmail，更强大、更安全、更多存储空间！ 立刻体验！ |
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In reply to this post by Charles Matthew Blaha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal So is this scaremongering or real? Has anybody on the list ever had a microscope degrade/die/rust to smithereens because they coupled it to an box-style humidified chamber? And has anyone ever used the Bioptechs rebuilding service for these reasons, and if so, what was found? Saludos Chris Dr Chris Wood Instituto de Biotecnología Universidad Nacional Autónoma de México Av. Universidad 2001 Col. Chamilpa Cuernavaca 62150 Morelos México On Tue, 13 Nov 2007 21:54:09 -0500, George McNamara <[hidden email]> wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hi Charles, > >Check out the Precautionary Note section of the web page >http://www.bioptechs.com/Products/Rebuild/rebuilding_service.html > > > > |
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Higdon, Michael |
Re: Microscope Stage Incubators
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In reply to this post by Charles Matthew Blaha
I actually worked on an Olympus IMT2 that was regularly bathed in saline. It took seven years for the focus mechanism to finally lockup with all the salt. I personally think the "acrylic-box" is the way to go! |
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Jacqueline Ross |
Re: Microscope Stage Incubators
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In reply to this post by Chris Wood-5
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Charles & others, We have Solent incubation (box-style) systems on two microscope systems, one an Olympus FV1000 confocal microscope and the other a Nikon TE2000E inverted microscope. We've had both systems for around 2 years without obvious rust issues at the moment. Both systems are multi-user. With these systems, the gas is humidified but it comes up to the working head of the microscope and of course your specimen is bathed in liquid but it's still a fairly small volume in our case. I've never noticed condensation dripping off the sides of the box and since it's not completely sealed, moisture does have a chance to dry out. In fact, the real risk of rust comes with the use of the dipping objectives on our confocal, which drip down buffer salts, etc. onto the condenser, etc. So it is important to clean that off to avoid rust. The reason we bought a box-style system for our inverted microscope was because when I had a look at all the available chamber-based systems, there seemed to be variable reports about how well they worked especially since we operate as a multi-user facility. Our systems are also used by people who don't need the incubation system but because we have the box structure rather than a chamber system, we don't have to keep taking things off and on. One other concern was that you seemed to need to buy other components such as objective heaters, etc. to avoid heat sinks/focus drift if you have a stage-top incubator whereas with the box-style, more of the system is heated and as long as you give sufficient time for things to equilibrate, I think it works quite well. However, auto-focus (as mentioned previously by Gabor) is still a useful thing to have with a box-style system. Kind regards, Jacqui Jacqueline Ross Biomedical Imaging Microscopist Biomedical Imaging Research Unit School of Medical Sciences Faculty of Medical & Health Sciences The University of Auckland Private Bag 92019 Auckland, NEW ZEALAND Tel: 64 9 373 7599 Ext 87438 Fax: 64 9 373 7484 http://www.health.auckland.ac.nz/biru/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Chris Wood Sent: 15 November 2007 11:44 To: [hidden email] Subject: Re: Microscope Stage Incubators Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal So is this scaremongering or real? Has anybody on the list ever had a microscope degrade/die/rust to smithereens because they coupled it to an box-style humidified chamber? And has anyone ever used the Bioptechs rebuilding service for these reasons, and if so, what was found? Saludos Chris Dr Chris Wood Instituto de Biotecnología Universidad Nacional Autónoma de México Av. Universidad 2001 Col. Chamilpa Cuernavaca 62150 Morelos México On Tue, 13 Nov 2007 21:54:09 -0500, George McNamara <[hidden email]> wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hi Charles, > >Check out the Precautionary Note section of the web page >http://www.bioptechs.com/Products/Rebuild/rebuilding_service.html > > > > |
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Jacqueline Ross |
Re: Microscope Stage Incubators
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi again, I just realised that when I mentioned about the humidified gas coming up to the working head, it may not be clear what I meant for those who aren't familiar with these Solent systems. The humidified gas in comes through narrow tubing up to the specimen rather than filling the entire box. Kind regards, Jacqui Jacqueline Ross Biomedical Imaging Microscopist Biomedical Imaging Research Unit School of Medical Sciences Faculty of Medical & Health Sciences The University of Auckland Private Bag 92019 Auckland, NEW ZEALAND Tel: 64 9 373 7599 Ext 87438 Fax: 64 9 373 7484 http://www.health.auckland.ac.nz/biru/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Jacqui Ross Sent: 15 November 2007 13:04 To: [hidden email] Subject: Re: Microscope Stage Incubators Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Charles & others, We have Solent incubation (box-style) systems on two microscope systems, one an Olympus FV1000 confocal microscope and the other a Nikon TE2000E inverted microscope. We've had both systems for around 2 years without obvious rust issues at the moment. Both systems are multi-user. With these systems, the gas is humidified but it comes up to the working head of the microscope and of course your specimen is bathed in liquid but it's still a fairly small volume in our case. I've never noticed condensation dripping off the sides of the box and since it's not completely sealed, moisture does have a chance to dry out. In fact, the real risk of rust comes with the use of the dipping objectives on our confocal, which drip down buffer salts, etc. onto the condenser, etc. So it is important to clean that off to avoid rust. The reason we bought a box-style system for our inverted microscope was because when I had a look at all the available chamber-based systems, there seemed to be variable reports about how well they worked especially since we operate as a multi-user facility. Our systems are also used by people who don't need the incubation system but because we have the box structure rather than a chamber system, we don't have to keep taking things off and on. One other concern was that you seemed to need to buy other components such as objective heaters, etc. to avoid heat sinks/focus drift if you have a stage-top incubator whereas with the box-style, more of the system is heated and as long as you give sufficient time for things to equilibrate, I think it works quite well. However, auto-focus (as mentioned previously by Gabor) is still a useful thing to have with a box-style system. Kind regards, Jacqui Jacqueline Ross Biomedical Imaging Microscopist Biomedical Imaging Research Unit School of Medical Sciences Faculty of Medical & Health Sciences The University of Auckland Private Bag 92019 Auckland, NEW ZEALAND Tel: 64 9 373 7599 Ext 87438 Fax: 64 9 373 7484 http://www.health.auckland.ac.nz/biru/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Chris Wood Sent: 15 November 2007 11:44 To: [hidden email] Subject: Re: Microscope Stage Incubators Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal So is this scaremongering or real? Has anybody on the list ever had a microscope degrade/die/rust to smithereens because they coupled it to an box-style humidified chamber? And has anyone ever used the Bioptechs rebuilding service for these reasons, and if so, what was found? Saludos Chris Dr Chris Wood Instituto de Biotecnología Universidad Nacional Autónoma de México Av. Universidad 2001 Col. Chamilpa Cuernavaca 62150 Morelos México On Tue, 13 Nov 2007 21:54:09 -0500, George McNamara <[hidden email]> wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hi Charles, > >Check out the Precautionary Note section of the web page >http://www.bioptechs.com/Products/Rebuild/rebuilding_service.html > > > > |
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