**5. Use of polscope**

120 Enhancing Success of Assisted Reproduction

does not improve the overall results.

**3. Use of PICSI** 

drops).

**4. Use of IMSI** 

pushing the needle tip close to the membrane opposite the puncture site, aspirating the cytoplasm at this point and releasing the sperm in the centre of oocyte [5]. This modification improves fertilization in oocyte-dependent activation failure, but its routine application

The cell surface hyaluronic acid (HA) binding glycoprotein is present in spermatozoa of different species including rat, mice, bull, and human [15]. The formation of hyaluronic-acid (HA)-binding sites on the sperm plasma membrane is one of the signs of sperm maturity. Various biochemical sperm markers indicate that human sperm bound to HA exhibit attributes similar to that of zona pellucida-bound sperm, including minimal DNA

PICSI Sperm Selection Device (Biocoat, Inc. Horsham, PA, USA) offers advantage in selecting sperm for ICSI. The PICSI device, a dish similar to ICSI dish, contains 3 microdots of hyaluronan hydrogel which need to be hydrated by media before ICSI. The prepared sperm sample is placed at the edge of the microdrop of PICSI dish. Mature, biochemically competent sperm bind to the hyaluronan where they can be isolated by the embryologist and used for ICSI **(Figure 3)**. The research supports that hyaluronan-bound PICSI-selected sperm are, in the vast majority of cases, more mature, exhibit less DNA damage, and have fewer chromosomal aneuploidies [17]. Further studies are needed to prove that use of PICSI

fragmentation, normal shape, and low frequency of chromosomal aneuploidies [16].

technique improves pregnancy rates and reduce the number of IVF miscarriages.

**Figure 3.** A is a graphical presentation of a PICSI dish. Each arrow is pointing to a dot containing hyaluronan. B and C are suggested arrangements for oocyte washing, PVP and ICSI drops (A, B and C are oocyte washing drops, P is PVP drop, a, b and c are hyaluronan dots and1, 2, 3, and 4 are ICSI

Intracytoplasmic morphologically selected sperm injection (IMSI) is examination of unstained spermatozoa at 6000 or higher magnification to select sperm with best morphology. It is based on a method of high magnification motile sperm organelle morphology examination (MSOME) [18]. It requires an inverted light microscope equipped The oocyte spindle can be imaged non-invasively based on the birefringence, an inherent optical property of highly ordered molecules, such as microtubules, as they are illuminated with polarized light. Polarized light microscopy has been applied to embryology for decades. A digital, orientation-independent polarized light microscope, the Polscope, has demonstrated the exquisite sensitivity needed to image the low levels of birefringence exhibited by mammalian spindles [26]. The Polscope, is used to protect the meiotic spindle from damage during ICSI. The oocytes having Polscope visualised spindle have higher fertilization rate. When the spindle is located at 0°-30° in relation to the first polar body, ICSI achieves highest fertilization rate [27]. The use of Polscope is still not widely practiced and further improvements are needed. Morphometric evaluation of the spindle through the Polscope is not consistent with confocal analysis. This suggests that the Polscope may still be a rather inefficient method for assessing the metaphase II spindle [28].
