**3. Use of PICSI**

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

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 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 drops).
