Reproductive Medicine

Reproductive Medicine

In vitro fertilization (IVF), Intracytoplasmic sperm injection (ICSI) and Intracytoplasmic morphologically-selected sperm injection (IMSI) are reproductive technologies for fertility treatment of humans and animals. All those methods have in common, that eggs (ova) are removed from a female and fertilized outside the female’s body (in vitro) with sperms. Fertilized eggs (zygotes) are then implanted into the same or another female’s uterus to establish pregnancy.

 

In vitro fertilization (IVF)

  • eggs are harvested and incubated
  • unselected sperm cells are added
  • both are left together (in vitro) for several days
  • healthy, mobile sperm cells actively fertilize the eggs
  • embryos are transferred into the uterus

Intracytoplasmic sperm injection (ICSI):

  • eggs are harvested and incubated
  • the oocyte is stabilized by a holding pipette
  • a glass micropipette is used to collect a single sperm
  • the unselected sperm cell is immobilized by cutting its tail with the point of the micropipette
  • the oocyte is pierced through the membrane (oolemma) and the sperm is directed to the inner part of the oocyte (cytoplasm)
  • the sperm is then released into the oocyte
  • cellular structures such as the zona pellucida and polar body of the egg cell must be clearly visible  

Intracytoplasmic morphologically-selected sperm injection (IMSI)

  • eggs are harvested and incubated
  • morphological selection of a healthy egg with an inverted microscope
  • the oocyte is stabilized by a holding pipette
  • semen analysis with an upright microscope
  • morphological selection of a healthy sperm cell with an inverted microscope with high magnifications, DIC contrast with oil immersion objectives
  • a glass micropipette is used to immobilize the selected sperm by cutting its tail
  • the oocyte is pierced through the membrane (oolemma) and the sperm is directed to the inner part of the oocyte (cytoplasm)
  • the sperm is then released into the oocyte
  • cellular structures such as the zona pellucida and polar body of the egg cell must be clearly visible
  • shape and vacuole count of the sperm cells must be assessed
     

For semen analysis and sperm preparation mainly upright microscopes are used. Male infertility is diagnosed by detailed semen analysis that look specifically at the total volume, the number of motile sperm per millilitre (ml) and sperm morphology. The pre-selection of the sperms take place in glass dishes, requiring a microscope with DIC or polarization contrast.  

Harvested cells are kept in culture dishes at body temperature. For examination and selection microscopes with heating stages are essential. First sortings are often done with compact stereo microscopes such as Stemi 508. Egg cells lie at the bottom of the culture dish. To assess their morphology, the observing objective needs to be close to them. That’s why inverted microscopes are used. They additionally offer ample space for micromanipulation. Also, for ICSI and IMSI the microscope systems must be exceptionally stable and vibration free. Special contrasting techniques allow to assess the morphology of unstained egg cells. Especially ICSI and IMSI require high resolution and high contrast. Documentation and co-observation are often helpful.  
 

Differential Interference Contrast (DIC)

DIC delivers high resolution images to visualize even the finest structures in cells. This method uses polarized light to detect differences in the refractory index in specimens. Because plastic does not preserve the polarization of light, DIC does not work with plastic dishes. 

PlasDIC

ICSI is often performed in plastic petri dishes, where DIC does not work. PlasDIC from ZEISS overcomes this limitation and delivers crisp relief contrast and an impressive three-dimensional image. 

Hoffmann Modulation Contrast (HMC)

HMC is the classic contrasting technique for unstained, low-contrast specimens. The improved HMC (iHMC) from ZEISS delivers a crisp relief contrast and reveals even finest structures in the cell nucleus, the nucleus shape and nucleoli. iHMC can be used with plastic dishes.

Polarization Contrast

The sperm selection for IMSI is often performed in glass dishes, where polarized light needs to be used.  

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ZEISS Microscope Systems for Assisted Reproductive Technologies

IVF, ICSI and IMSI

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Recommended Products for Reproductive Medicine

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Axio Lab.A1 has been designed for daily laboratory applications. All applications in brightfield, darkfield, phase contrast or fluorescence can be performed readily. The upright microscope places strong emphasis on ergonomic features.

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ZEISS Stemi 508

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Stemi 508 is a compact stereomicroscope, that fits in standard Laminar Flow Boxes. The Greenough system delivers brilliant three-dimensional images. The large working distance offers ample space for handling and sorting egg cells. The double spot gooseneck illumination can be easily adjusted to always deliver optimal contrast. The operation of the microscope is easy and ergonomic. Stemi 508 allows quick success monitoring of injected egg cells.

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ZEISS SteREO Discovery.V8

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SteREO Discovery.V8 is your modular stereomicroscope with an extensive range of accessories. The stereoscopic image helps you to better observe, understand, and manipulate your specimens. The large working distance offers ample space for handling and sorting egg cells. SteREO Discovery.V8 allows quick isolation of eggs before fertilization and then assessing the growing embryos.

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ZEISS Axio Vert.A1

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Axio Observer is the most stable inverted microscope, combining IMSI and ICSI in a single microscope. The modular stand can be motorized, equipped with numerous heating and incubation options and features several fixation points for micromanipulators. Additionally to DIC, PlasDIC and iHMC transmitted light darkfield is available. Axio Observer allows the sperm cell to be injected into the egg cell immediately after selection. The large field of view provides a clear picture of the egg and sperm cell with the injectors. Incubation can then be continued in an incubator with no problem at all. The ability to switch quickly from high to medium magnification is particularly beneficial for the subsequent assessment of the embryo.  

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