Re: [ccp4bb] sealing slides on VDX plates?
I have used melted Vaseline and a 10mL Erlenmeyer flask. The mouth of the Erlenmeyer is the same size as the opening of the VDX plate. Dip flask mouth in melted Vaseline, then press the mouth of the flask onto the opening of the VDX plate. - Original Message - > From: "Dima Klenchin" > To: CCP4BB@JISCMAIL.AC.UK > Sent: Friday, November 18, 2011 11:55:07 AM > Subject: [ccp4bb] sealing slides on VDX plates? > Hello, > > I wonder what everyone is using for sealing hanging drop slides on VDX > plates? For the most part, we paraffin oil but I am unhappy with it > because > it is too think and too frequently there is break in the oil and the > drop > dries too much. I find vacuum grease to be not terribly practical > because > it takes too much time - particularly when the well needs to be opened > (seeding, modifying well content, etc). > > In ideal world I would like to find a much thicker oil that 1) > contains as > little volatiles as paraffin oil, and 2) allows no more water vapor > diffusion through it than paraffin oil. Hampton suggests Cargille > immersion > oils for this purpose but the MSDS for these oils states that they > have > slight odor, so I am a bit concerned with unknown volatiles getting > into > crystallization drop. > > So, what do you like to use? Thanks much, > > - Dima -- Kay Perry NE-CAT 9700 S. Cass Ave. Bldg. 436 E007 Argonne, IL 60439 630-252-0692
Re: [ccp4bb] sealing slides on VDX plates?
Haven't done this for a while, but what we used to do was mix Vaseline plus mineral oil (both purchased for cheap at the local drugstore), and then apply it using a 10 ml syringe with a pipet tip attached. We used the mixture of Vaseline + mineral oil because Vaseline alone is too viscous (unless you want to develop Popeye forearms). Heat a jar of Vaseline in a hot water bath (NOT OVER A FLAME--it's flammable!) until it melts. Mix in about 10-20% mineral oil by volume (you can play with this ratio until you find the viscosity you like); then draw the mixture up into syringes while still liquid. The procedure a little messy, so fill a lot of syringes at once and you're set for years. Pat On 18 Nov 2011, at 12:55 PM, Dima Klenchin wrote: > Hello, > > I wonder what everyone is using for sealing hanging drop slides on VDX > plates? For the most part, we paraffin oil but I am unhappy with it because > it is too think and too frequently there is break in the oil and the drop > dries too much. I find vacuum grease to be not terribly practical because it > takes too much time - particularly when the well needs to be opened (seeding, > modifying well content, etc). > > In ideal world I would like to find a much thicker oil that 1) contains as > little volatiles as paraffin oil, and 2) allows no more water vapor diffusion > through it than paraffin oil. Hampton suggests Cargille immersion oils for > this purpose but the MSDS for these oils states that they have slight odor, > so I am a bit concerned with unknown volatiles getting into crystallization > drop. > > So, what do you like to use? Thanks much, > > - Dima
Re: [ccp4bb] sealing slides on VDX plates?
The is probably the cleanest and fastest way of greasing plates. Take a 15ml Falcon tube and unscrew the cap. You will notice that the cap is exactly the width as the well of the VDX plate. Using a fairly wide bored metal syringe needle, pierce 8 holes into the cap from the outside, along the top edge of the cap. Fill the Falcon tube with Vaseline and screw the cap back onto the tube. Loosen the cap about 2 to 2.5 turns, just before it comes of the tube. Stick the tube in a heating block, wait for the Vaseline to melt. When you now invert the tube with the liquid Vaseline, it should not come running out, but if you press the top of the cap against the well of the VDX plate, you will see that a small amount of Vaseline is let out through the small holes. If you lift the tube off the well, air will then go into the tube and you can press the cap against the next well. Continue until you're. Should take no more than a few seconds to do the whole plate. Cheers, Chris Browning -- Dr. Christopher Browning Post-Doctor to Prof. Petr Leiman EPFL BSP-416 1015 Lausanne Tel: 0041 (0) 02 16 93 04 40
Re: [ccp4bb] sealing slides on VDX plates?
I remember a technique that used warmed Vaseline (liquefied) in a shallow pan. People would invert the plate and dip it into the liquefied Vaseline, then flip it over to dry. I suppose one could use masking tape to cover the outer edge of the plate to keep it Vaseline free. I am not sure how it would work with cryschem (24-well sitting drop) plates. Good luck! Bryan -- Confidentiality Notice: This message is private and may contain confidential and proprietary information. If you have received this message in error, please notify us and remove it from your system and note that you must not copy, distribute or take any action in reliance on it. Any unauthorized use or disclosure of the contents of this message is not permitted and may be unlawful. -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of malho...@miami.edu Sent: Friday, November 18, 2011 1:26 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] sealing slides on VDX plates? Vaseline (pure, with no additives) from CVS/Walgreens in a 10 ml syringe and a pipette tip (wide bore) also works very well. On 11/18/11 12:58 PM, Bosch, Juergen wrote: > Dow Corning Heavy Vacuum Grease, a 10 ml syringe and a shorter cut 200 > µl tip attached to the syringe. > > Jürgen > > On Nov 18, 2011, at 12:55 PM, Dima Klenchin wrote: > >> Hello, >> >> I wonder what everyone is using for sealing hanging drop slides on VDX >> plates? For the most part, we paraffin oil but I am unhappy with it >> because >> it is too think and too frequently there is break in the oil and the >> drop >> dries too much. I find vacuum grease to be not terribly practical >> because >> it takes too much time - particularly when the well needs to be opened >> (seeding, modifying well content, etc). >> >> In ideal world I would like to find a much thicker oil that 1) >> contains as >> little volatiles as paraffin oil, and 2) allows no more water vapor >> diffusion through it than paraffin oil. Hampton suggests Cargille >> immersion >> oils for this purpose but the MSDS for these oils states that they have >> slight odor, so I am a bit concerned with unknown volatiles getting into >> crystallization drop. >> >> So, what do you like to use? Thanks much, >> >> - Dima > -- Arun Malhotra Phone: (305) 243-2826 Associate ProfessorLab: (305) 243-2890 Dept. of Biochemistry& Molecular Biology Fax: (305) 243-3955 University of Miami School of Medicine PO Box 016129 E-Mail: malho...@miami.edu Miami, FL 33101 Web: http://structure.med.miami.edu
Re: [ccp4bb] sealing slides on VDX plates?
Vaseline (pure, with no additives) from CVS/Walgreens in a 10 ml syringe and a pipette tip (wide bore) also works very well. On 11/18/11 12:58 PM, Bosch, Juergen wrote: Dow Corning Heavy Vacuum Grease, a 10 ml syringe and a shorter cut 200 µl tip attached to the syringe. Jürgen On Nov 18, 2011, at 12:55 PM, Dima Klenchin wrote: Hello, I wonder what everyone is using for sealing hanging drop slides on VDX plates? For the most part, we paraffin oil but I am unhappy with it because it is too think and too frequently there is break in the oil and the drop dries too much. I find vacuum grease to be not terribly practical because it takes too much time - particularly when the well needs to be opened (seeding, modifying well content, etc). In ideal world I would like to find a much thicker oil that 1) contains as little volatiles as paraffin oil, and 2) allows no more water vapor diffusion through it than paraffin oil. Hampton suggests Cargille immersion oils for this purpose but the MSDS for these oils states that they have slight odor, so I am a bit concerned with unknown volatiles getting into crystallization drop. So, what do you like to use? Thanks much, - Dima -- Arun Malhotra Phone: (305) 243-2826 Associate ProfessorLab: (305) 243-2890 Dept. of Biochemistry& Molecular Biology Fax: (305) 243-3955 University of Miami School of Medicine PO Box 016129 E-Mail: malho...@miami.edu Miami, FL 33101 Web: http://structure.med.miami.edu
Re: [ccp4bb] sealing slides on VDX plates?
Dow Corning Heavy Vacuum Grease, a 10 ml syringe and a shorter cut 200 µl tip attached to the syringe. Jürgen On Nov 18, 2011, at 12:55 PM, Dima Klenchin wrote: Hello, I wonder what everyone is using for sealing hanging drop slides on VDX plates? For the most part, we paraffin oil but I am unhappy with it because it is too think and too frequently there is break in the oil and the drop dries too much. I find vacuum grease to be not terribly practical because it takes too much time - particularly when the well needs to be opened (seeding, modifying well content, etc). In ideal world I would like to find a much thicker oil that 1) contains as little volatiles as paraffin oil, and 2) allows no more water vapor diffusion through it than paraffin oil. Hampton suggests Cargille immersion oils for this purpose but the MSDS for these oils states that they have slight odor, so I am a bit concerned with unknown volatiles getting into crystallization drop. So, what do you like to use? Thanks much, - Dima .. Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-2926 http://web.mac.com/bosch_lab/
[ccp4bb] sealing slides on VDX plates?
Hello, I wonder what everyone is using for sealing hanging drop slides on VDX plates? For the most part, we paraffin oil but I am unhappy with it because it is too think and too frequently there is break in the oil and the drop dries too much. I find vacuum grease to be not terribly practical because it takes too much time - particularly when the well needs to be opened (seeding, modifying well content, etc). In ideal world I would like to find a much thicker oil that 1) contains as little volatiles as paraffin oil, and 2) allows no more water vapor diffusion through it than paraffin oil. Hampton suggests Cargille immersion oils for this purpose but the MSDS for these oils states that they have slight odor, so I am a bit concerned with unknown volatiles getting into crystallization drop. So, what do you like to use? Thanks much, - Dima
