Biomedical subjects

A Clement-Sengewald

Publications and source records attributed to A Clement-Sengewald.

7 recordsLinked to original sources

PubMed2002

Noncontact, laser-mediated extraction of polar bodies for prefertilization genetic diagnosis.

PURPOSE: We tested an entirely noncontact polar body-extraction method using an ultraviolet laser beam for laser zona drilling and a near infrared laser beam for polar body (PB) trapping and extraction. METHODS: A hole was drilled into the zona pellucida of an oocyte. Then, the PB was trapped with optical tweezers and dragged through the drilled hole. RESULTS: Bovine first PBs could be extracted in 49 out of 63 oocytes (78%) using this method. In human oocytes, PB extraction was successfully demonstrated, which however was more time consuming. A number of extracted PBs were dried on a special membrane, circumcised with the laser microbeam, and successfully catapulted into the lid of a microfuge tube (laser pressure catapulting). CONCLUSIONS: This solely laser-mediated extraction method allows convenient procurement of PBs without the danger of contamination and is a promising approach that might replace standard micromanipulation methods in the future.

Animals↗

PubMed2000

Laser microdissection as a new approach to prefertilization genetic diagnosis.

The genetic status of oocytes can be determined by polar body (PB) analysis. Following PB extraction, a genetic evaluation is performed. As each PB contains the complementary genetic material of the oocyte, PB analysis reveals information about its genetic status. Genetically altered oocytes may then be excluded from in vitro fertilization. The aim of our study was to evaluate laser microdissection as a tool for PB extraction purposes. Compared to the PB extraction with a sharp-ending pipette only, we could show that laser microdissection of the zona pellucida (laser zona drilling) with a UV-A laser and subsequent extraction with a blunt-ending pipette decreases the degeneration rate of oocytes. It is shown that laser pressure catapulting of extracted PB enables their contact-free transfer into tubes, thus decreasing the risk of contamination for further analysis.

Cell Separation↗

PubMed1996

Fertilization of bovine oocytes induced solely with combined laser microbeam and optical tweezers.

PURPOSE: Our purpose was to show that fertilization of oocytes can be obtained solely by laser light-mediated manipulation of gametes. METHOD: A small channel was drilled into the zona pellucida of bovine oocytes using an ultraviolet (UV)-laser microbeam. Highly diluted cattle sperm were not able to fertilize the laser drilled oocytes. RESULTS: Fertilization was achieved only when three to five cattle sperm were trapped with optical tweezers and inserted directly through the laser drilled hole into the perivitelline space. After 20 hr, 3 of 79 (3.8%) oocytes revealed two pronuclei and a sperm tail within their cytoplasm. Cattle sperm are difficult to catch. Therefore, the gametes had to remain for about 20 min in room atmosphere, which might be the reason for the low fertilization results. CONCLUSIONS: The results indicate that a combined UV-laser microbeam and optical tweezers trap can be used successfully for "noncontact" microinsemination procedures.

Animals↗

PubMed1994

Catch and move--cut or fuse.

Still almost unbelievable, but true: light exerts force. With these forces it is indeed possible to catch and move cells or small particles and microsurgically to process them without any mechanical contact. As if by magic, objects are moved via focused laser light.

Animals↗

PubMed1994

Zona drilling and sperm insertion with combined laser microbeam and optical tweezers.

A combined UV-laser microbeam and optical-tweezers trap was used to perform laser zona drilling and subzonal insemination in cattle. Using a precisely focused UV-laser microbeam, a small channel of about 10 microns in diameter was drilled into the zona pellucida. With a three-dimensional optical-tweezers trap, a single sperm was caught and transported through the laser-drilled hole directly into the perivitelline space. Furthermore, the sperm was brought into close contact with the oolemma to facilitate sperm-oocyte fusion. Using the laser-microscope system, noncontact, entirely sterile, and highly selective micromanipulation of gametes can be achieved with no need for mechanical microtools. Laser micromanipulation seems to be less detrimental to the gametes and is comparatively is easy to perform. Thus, the combined UV-laser microbeam and optical tweezers trap may be a helpful tool for IVF procedures.

Animals↗

PubMed1992

[Embryo cloning in domestic animals].

The cloning of livestock is performed by the nuclear transfer of early embryonic stages into prepared oocytes in order to obtain a high number of genetic identical animals. As the most important technical steps there are maturation and enucleation of the oocytes, isolation of single blastomeres or karyoplasts of the donor embryo, transfer of the nucleus-containing membrane vesicle under the zona pellucida of the recipient cell, and fusion of the recipient cell and blastomere or karyoplast. Until now, the largest clone which is known exists of seven bulls. The efficiencies of the particular methodical steps have to be improved. More knowledge of the activation of oocytes, nucleus differentiation and availability of determined cell cycle-stages of mitosis is required. The combination of embryo cloning, cryopreservation of embryos and non-surgical embryo transfer is required for basic research and animal breeding.

Animals↗

PubMed1989

Electrofusion parameters for mouse two-cell embryos.

We studied electrofusion of mouse two-cell embryos in order to define parameters which would result in a high yield of fused embryos. Various cell alignment times (from <10 to >60 s) and alternating current percentages (2 to 100%) were examined. The fusion parameters tested were the number of fusion pulses (1-9), pulse length (30-90 mus) and pulse strength (0.50-1.79 kV/cm). Furthermore different combinations of these three parameters were tested. In addition the influence of several embryo culture media on the fusion rates was examined. The results show that the fusion rate of the embryos increases with shorter alignment and higher percentages of the alternating current. The highest fusion rate (95%) was obtained by use of one pulse with a duration of 70 mus and a field strength of 0.60-0.79 kV/cm. The survival rate of the embryos was best if Whitten Medium was used before and after the fusion pulses. The fusion of two-cell stages results in tetraploid embryos which can serve as models for studies in polyploid cells.

Journal Article↗