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Jos? Rosales, LIVV (Venezuela) said on 10 October 2004
"I agree. If you use a Co2 incubator in order to warm an HEPES bufered medium you have to close cap. I tested pH value in Flushing medium (Medicult) warming open overnigth in the incubator and it was near 7,30."
Reply
KELLY HANSHEW, BRYN MAWR HOSPITAL said on 10 October 2004
"We warm our HEPES buffered medium overnight in a room air 37 degree C dry oven. Since it is HEPES buffered it should not be exposed to increased CO2 conditions because it will alter the pH of the media. We try to minimize the amount of HEPES media that is transferred with our sperm or oocytes; again due to pH alteration. If you use oil overlay you should be able to manipulate the zygotes and embryos successfully without the use of HEPES if you only have them out of the incubator for a short period of time."
Reply
Levent Keskintepe, Seer Institute said on 10 October 2004
"I do not think that you need to equilibrate any media which contains HEPES in it. It will be acidic if one tries with CO2. HEPES containing media should be warmed before used."
Reply
Dr.D'Pankar Banerji, Ideal Fertility and IVF Center,jabalpur,Ind said on 10 October 2004
"Any media which is buffered by bicarbonate must be equilibrated by + 37 deg C + 5 % CO2. even though the media's pH is further stabilized by HEPES.
The gamete handling meda ( bicarbonate bufferd ) are added with HEPES may be due to absorb the shock of pH disturbance due to its prolong exposure to
atmospheric air .
The actual IVF media are not stabilized by HEPES because it may cause ionic changes in embryos and the chances of exposure to atmosphere is less.
Thanks"
Reply
James Stanger, Hunter IVF/FertAid said on 10 October 2004
"I believe it should be equilbrated in a standard incubator. The pH has been adjusted to 7.4 with HEPES and usually 5mM Bicarbonate.If you put it in a 5% CO2 incubator the pH will fall below 7.0
Best wishes."
Reply
Xavier Blum, Innaifest-Ecuador said on 10 October 2004
"Only need 37 ?C , not CO2 , if you use CO2 incubator close cap."
Reply