Questions related to ICSI
Hello everyone. I would like to discuss my ICSI procedure. I´ve been learning ICSI method for a year within my thesis, that focused on interspecific ICSI (specifically mouse oocytes and goat sperm). My problem is, that i can not reach satisfying numbers of viable injected oocytes. A lot of oocytes break and flow out during an injection. If they survive, some of them die later during cultivation in incubator. I hope you can understand, that I´m still a rookie, but I hope you can give me some good advices. The procedure is as follows:
- PMSG stimulation
- hCG stimulation (48 h after PMSG)
- Isolation of oocytes (16-17h after hCG)
- --- for isolation: M2 medium (10ml M2-HEPES + 10 mg BSA, stored in 4°C, filtered, heated to 36,8°C 1/2 h before use)
- Removal of cumulus cells - in drops of hyaluronidase (2 mg HYA + 2 ml PBS, heated before use)
- --- I have always been careful about this step for limited time (few seconds) that oocytes spend in drops. If cumulus cells have been removed, oocytes are washed in fresh drops of same M2 medium. Then oocytes are moved to drops of culture medium - usually MEM (40 mg BSA + 10 ml MEM + Na pyruvate gentamicin 100 μg).
- Oocyte "rest" - in 4well in MEM medium in incubator with 5% CO2 . Recovering from isolation for 1/2 h.
- ICSI - oocytes are moved from MEM to M2 medium. First in fresh drops outside, then M2 drops in manipulation dish (covered with mineral oil).
- Sperm that is used for this procedure is frozen, so it is thawed and moved to M2 medium (centrifuged and washed from cyroprotectants)
- Piezo system is used for this procedure. Injection pipette contains minimal amount of mercury. Pulses regulation depending on oocytes condition. Trying to use the bigger pulse for breaking through ZP and smaller pulse to get into cytoplasm. Than pull injection pipette slowly from oocyte.
Could you please tell me, if there is any critical thing in this procedure that could cause this problem. My master uses same protocol (except that he is a pro with many years of experience). I really understand that it is most likely my "hands" that make mistakes. So this is a little bit challenge for me, but I believe that you can understand that everyone had to start and learn.
Thank you all for your answers :)
I have values for both variables over time, I did a simple correlation between them for each time point, and the results are very variable, and If I do a global correlation (data from all times together), the correlation is not true. I have been looking for what is the best procedure, and I have found information about autoregression but this method is for correlating any variable along the time, but not the correlation between two variables along the time. Anybody could help me?
We are trying to create a transgenic mice in our lab for the first time. We currently have a microinjector that we use for ICSI or biopsies, which is fitted with mineral oil injectors, to control the pipettes. In all the papers I´ve read about this, people use air pressure-controlled injectors to inject a small volume into the embryo. Has any of you attempted something like this using the oil injector, or is it necessary to have the air injector? Thank you!
more often we found degeneration in oocytes after ICSI ? was just curious to know what can be the reasons apart from technical error?
I would like to model the human oocyte flowing into a micro-channel. I would like to find information about the density (weight) of the human oocyte. I would like to know if the density of the human oocyte is bigger than the density of the solution used in intra cytoplasmic sperm injection procedure. When the human oocyte is immersed into the solution, does it go down to the bottom of the micro-channel because of gravitation, or it is floating freely into the solution?
We previously did ICSI with this patients sperm and got 6 blastocysts which were tested normal using array CGH, yet no pregnancy outcome
Hi, I'm looking at embryonic quality score data for IVF/ICSI (multiple embryos per mother), which are categorical and ordinal. I want to compare 2 groups: 1 has had a certain type of surgery, the other hasn't.
The only way I know how to analyze categorical data is chi-square or fisher's exact. However, that doesn't take into account that the dependent variable is ordinal, and also leaves the problem that not all data are independent (multiple embryos from the same parents).
How could I best analyze this? (also preferably in SPSS)
Any help would be greatly appreciated.
32y old patient, candidate for frozen-thawed embryo transfer. Endometrium seemed resistant to estradiol validate, from 4 to 8 mg/day, and to estradiol patch, from 50 to 100 mug each 2 days.
I am developing a robotic system for cell injection that could be applied for ICSI. I would like to choose an IT platform and a language for programming. What kind of software is suitable for real time programming and control of different hardware devices as piezo-actuators, digital camera, microscope, micro-pumps. There are robotic toolbox, modules for image processing and many other modules for automation and control of robotic systems in real time in Labview and Matlab. I am not sure which language is better. In Matlab I can buy a specific hardware for control in opposite of Matlab, par example. The other possibility is to use linux OS, Qt4 programming language and OpenCV vision library as open source technology. The last option is to use Microsoft Visual Studio as a programming environment and selecting any programming language as C++ or C#. Could you please give me advice on what kind of a programming technology is best suited for development of software applications in real time? Thanks.
Could a frozen tissue saved from adjacent testis tumor jeopardize future offspring in those indiviiduals who need to undego to bilateral orquiectomy due to a TU?
Any guess and or suggestion?
It's generally thought that mtDNA is only maternally transmitted. But is there any evidence at all for paternal mtDNA transmission? What about during ICSI? And if so, what are the implications? Does it change what we can say about mitochondrial "Eve."?