Mice are ideal models for ICSI experiments because of the ease of culturing eggs/embryos in vitro and the availability of ample genetic information. Unfortunately, it has been extremely difficult. In this study we improved the mouse ICSI method by using a new holding pipette that was made of two pipettes such that one pipette was pulled and heat merged into the other one. The outer pipette ...

mice are ideal models for icsi experiments because of the ease of culturing eggs/embryos in vitro and the availability of ample genetic information. unfortunately, it has been extremely difficult. in this study we improved the mouse icsi method by using a new holding pipette that was made of two pipettes such that one pipette was pulled and heat merged into the other one. the outer pipette had ...

Mouse oocytes were placed in Hepes-T6 containing 7.5 μg/ml cytochalasin B and 5μg/ml Hoechst prior to enucleation, and enucleation was performed on oocytes using the manipulator. The oocyte held in place by gentle suction of a holding pipette (oocyte diameter approx.70 μm). The metaphase spindle (arrow) is aspirated into an enucleation pipette (1).

The focal extracellular potential (FEP) described in this study is an electrophysiological signal related to the transmembrane potential (V(m)) of cardiac myocytes that avoids the mechanical fragility, interference with contraction, and intracellular contact associated with conventional whole cell recording. One end of a frog ventricular myocyte was secured into a glass holding pipette. The FEP...

Generating transgenic mice using fertilized egg cells Purified experimental DNA is directly injected into the male pronucleus of a fertilized mouse egg using a microscope-mounted manipulator and microneedle. The three essential instruments for embryo injection are an inverted microscope with stage magnification of 400X, a micromanipulator-mounted negative pressure holding pipette (left) and a m...

BACKGROUND/AIM Obtaining human embryonic stem cell lines has so far involved destroying the embryos. This has given rise to ethical concerns and is not permitted in most countries. This investigation tested whether removing multiple cells from blastocysts might allow continued embryonic development. MATERIALS AND METHODS A total of 40 blastocysts from a black mouse strain were biopsied. The m...

The reversibility and cation selectivity of the K(+)-Cl(-) cotransporter (KCC), which normally extrudes Cl(-) out of neurons, was investigated in dissociated lateral superior olive neurons of rats using the gramicidin perforated patch technique. Intracellular Cl(-) activity (alpha[Cl(-)](i)) was maintained well below electrochemical equilibrium as determined from the extracellular Cl(-) activit...

Effects of intracellularly perfused ATP, and extracellularly applied cyanide and 2-deoxy-D-glucose, on fast and slow Ca2+ channel currents of isolated single vascular smooth muscle cells were investigated by a whole-cell voltage-clamp method combined with an intracellular perfusion technique. Single smooth muscle cells were prepared by collagenase treatment from guinea pig small mesenteric arte...