F. Morel

Interindividual variations in the disomy frequencies of human spermatozoa and their correlation with nuclear maturity as evaluated by aniline blue staining

OBJECTIVE To evaluate the importance of interindividual variations in the disomy frequencies of human sperm and their possible correlation with the principal semen parameters. DESIGN Prospective randomized analysis of sperm nuclei by fluorescence in situ hybridization and analysis of semen parameters. SETTING University-based laboratory for reproductive biology. PATIENT(S) Fifty-seven human ejaculates selected at random from a population of men undergoing semen analysis. INTERVENTION(S) Semen specimens were analyzed, and sperm samples were prepared for fluorescence in situ hybridization. MAIN OUTCOME MEASURE(S) Semen parameters, including necrozoospermia, global motility, sperm concentration, multiple abnormalities index, and teratozoospermia were evaluated, aniline blue staining was completed, and disomy frequencies for chromosomes 8, 15, 18, X, and Y were determined using fluorescence in situ hybridization. RESULT(S) Noticeable differences in disomy frequencies between individuals were observed, and these frequencies were correlated with the degree of nuclear maturity. CONCLUSION(S) We hypothesize that the positive correlation can be explained by an abnormality of chromosomal segregation at the time of meiosis that would cause disturbances during the transition of nucleoprotein or by one or several premeiotic abnormalities of chromatin that would perturb both the meiotic process and the construction of definitive proteins.

First test results Of MIMOSA-26, a fast CMOS sensor with integrated zero suppression and digitized output

The MIMOSA pixel sensors developed in Strasbourg have demonstrated attractive features for the detection of charged particles in high energy physics. So far, full-size sensors have been prototyped only with analog readout, which limits the output rate to about 1000 frames/second. The new MIMOSA 26 sensor provides a 2.2 cm2 sensitive surface with an improved readout speed of 10,000 frames/second and data throughput compression. It incorporates pixel output discrimination for binary readout and zero suppression micro-circuits at the sensor periphery to stream only fired pixel out. The sensor is back from foundry since february 2009 and has being characterized in laboratory and in test beam. The temporal noise is measured around 13-14 e- and an operation point corresponding to an efficiency of 99.5±0.1 % for a fake rate of 10-4 per pixel can be reached at room temperature. MIMOSA 26 equips the final version of the EUDET beam telescope and prefigures the architecture of monolithic active pixel sensors (MAPS) for coming vertex detectors (STAR, CBM and ILC experiments) which have higher requirements. Developments in the architecture and technology of the sensors are ongoing and should allow to match the desired readout speed and radiation tolerance. Finally, the integration of MAPS into a micro-vertex detector is addressed. A prototype ladder equipped, on both sides, with a row of 6 MIMOSA 26-like sensors is under study, aiming for a total material budget about 0.3% X0.

Molecular analysis of the chromosomal equipment in spermatozoa of a 46, XY, t(7;8) (q11.21;cen) carrier by using fluorescence in situ hybridization.

Abstract The meiotic segregation of a balanced reciprocal translocation (7;8) (q11.21;cen) was analysed by interphase fluorescence in situ hybridization (FISH) on carrier spermatozoa. A dual interphase FISH technique was applied to 34527 decondensed sperm heads with chromosome-7- and chromosome-8-specific alpha-satellite probes. Analysis with such probes was possible according to the cytogenetic characteristics of these translocation breakpoints, which implied a centromeric breakpoint. The majority of the analysed nuclei (56.70%) showed normal (30.40%) or balanced (26.30%) chromosomal equipment resulting from alternate segregation during meiosis. A total of 14935 spermatozoa (43.26%) was unbalanced with a predominance of gametes resulting from adjacent-I (25.10%) or adjacent-II (11.10%) segregation ; such gametes could produce partial mono- or trisomies at term. The frequency of analysed cells resulting from a 3:1 segregation, which could induce complete mono- and trisomies at term, was 7.06%; 0.04% of scored cells were diploid. The same dual-FISH technique was carried out either with chromosome-15- and chromosome-18-specific probes or with gonosome-specific probes, in order to detect a possible interchromosomal effect. A significant increase of disomic18 spermatozoa was observed in the carrier. Such studies are not yet frequent. Multicolour-FISH seems a rapid and accurate tool for direct analyses of spermatogenetic segregation mechanisms in a carrier of balanced chromosomal abnormalities and provides interesting information for characterizing the possible risks for the offspring.

Estimation of aneuploidy levels for 8, 15, 18, X and Y chromosomes in 97 human sperm samples using fluorescence in situ hybridization

OBJECTIVE To estimate the mean frequency of aneuploidy levels of chromosomes 8, 15, 18, X, and Y in human sperm, while minimizing the effect of individual factors by analyzing sperm samples from a large set of patients. DESIGN Prospective randomized analysis of sperm nuclei by fluorescence in situ hybridization. SETTING University-based laboratory for reproductive biology. PATIENT(S) One hundred two patients with a large distribution of sperm parameters, randomly selected from volunteers who had presented seeking a semen analysis. INTERVENTION(S) The sperm samples were prepared for fluorescence in situ hybridization. MAIN OUTCOME MEASURE(S) The disomy frequencies for chromosomes 8, 15, 18, and sex chromosomes were determined using fluorescence in situ hybridization. RESULT(S) The mean frequencies of disomy for autosomes were 0.18% for chromosome 8, 0.06% for chromosome 15, 0.2% for chromosome 18, and 0.24% for gonosomes (XX, 0.04%; YY, 0.05%; XY, 0.15%). CONCLUSION(S) This study confirms other previous evaluations on restricted numbers of patients. Our results seem to confirm a relative equiprobability of disomy frequencies concerning the different chromosomal pairs during male meiosis.

Selective mitochondrial DNA degradation following double-strand breaks

Mitochondrial DNA (mtDNA) can undergo double-strand breaks (DSBs), caused by defective replication, or by various endogenous or exogenous sources, such as reactive oxygen species, chemotherapeutic agents or ionizing radiations. MtDNA encodes for proteins involved in ATP production, and maintenance of genome integrity following DSBs is thus of crucial importance. However, the mechanisms involved in mtDNA maintenance after DSBs remain unknown. In this study, we investigated the consequences of the production of mtDNA DSBs using a human inducible cell system expressing the restriction enzyme PstI targeted to mitochondria. Using this system, we could not find any support for DSB repair of mtDNA. Instead we observed a loss of the damaged mtDNA molecules and a severe decrease in mtDNA content. We demonstrate that none of the known mitochondrial nucleases are involved in the mtDNA degradation and that the DNA loss is not due to autophagy, mitophagy or apoptosis. Our study suggests that a still uncharacterized pathway for the targeted degradation of damaged mtDNA in a mitophagy/autophagy-independent manner is present in mitochondria, and might provide the main mechanism used by the cells to deal with DSBs.

A New Spatiotemporal CMOS Imager With Analog Accumulation Capability for Nanosecond Low-Power Pulse Detections

High-speed cameras use the interesting performances of CMOS imagers that offer advantages in on-chip functionalities, system power reduction, cost, and miniaturization. The FAst MOS Imager (FAMOSI) project consists in reproducing the streak camera functionality with a CMOS imager. In this paper, a new imager called FAMOSI 2, which implements an electronic shutter and analog accumulation capabilities inside the pixel, is presented. With this kind of pixel and the new architecture for controlling the integration, FAMOSI 2 can work in repetitive mode for low light power and in single shot mode for higher light power. This repetitive mode utilizes an analog accumulation to improve the sensitivity of the system with a standard n-well/psub photodiode. The characterization has been realized in single shot mode to optimize the accumulation mode. The prototype has been fabricated in the Austriamicrosystems 0.35-mum CMOS process. The chip is composed of 64 columns times 64 rows of pixels. The pixels have a size of 20 mum times 20 mum and a fill factor of 47%

Radiation tolerance of a column parallel CMOS sensor with high resistivity epitaxial layer

CMOS Monolithic Active Pixel Sensors (MAPS) demonstrate excellent performances in the field of charged particle tracking. A single point resolution of 1–2 μm and a detection efficiency close to 100% were routinely observed with various MAPS designs featuring up to 106 pixels on active areas as large as 4 cm2[1]. Those features make MAPS an interesting technology for vertex detectors in particle and heavy ion physics. In order to adapt the sensors to the high particle fluxes expected in this application, we designed a sensor with fast column parallel readout and partially depleted active volume. The latter feature was expected to increase the tolerance of the sensors to non-ionizing radiation by one order of magnitude with respect to the standard technology. This paper discusses the novel sensor and presents the results on its radiation tolerance.

MISTRAL & ASTRAL: two CMOS Pixel Sensor architectures suited to the Inner Tracking System of the ALICE experiment

A detector, equipped with 50 μm thin CMOS Pixel Sensors (CPS), is being designed for the upgrade of the Inner Tracking System (ITS) of the ALICE experiment at LHC. Two CPS flavours, MISTRAL and ASTRAL, are being developed at IPHC aiming to meet the requirements of the ITS upgrade. The first is derived from the MIMOSA28 sensor designed for the STAR-PXL detector. The second integrates a discriminator in each pixel to improve the readout speed and power consumption. This paper will describe in details the sensor development and show some preliminary test results.

Intermediate Digital Monolithic Pixel Sensor for the EUDET High Resolution Beam Telescope

A high resolution beam telescope, based on CMOS Monolithic Active Pixels Sensors (MAPS), is being developed under the EUDET collaboration, a coordinated detector R&D program for a future international linear collider. A very good spatial resolution < 5 mum, a fast readout time of 100 mus for the whole array (136 times 576 pixels) and a high granularity can be obtained with this technology. A recent fast MAPS chip, designed in AMS CMOS 0.35 mum Opto process with 14 mum epitaxial layer and called MIMOSA22, was submitted to foundry. MIMOSA22 has an active area of 26.5 mm2 with a pixel pitch of 18.4 mum arranged in an array of 576 rows by 136 columns where 8 columns have analog test outputs and 128 have their outputs connected to offset compensated discriminator stages. The pixel array is divided in seventeen blocks of pixels, with different amplification gain, diode size, pixel architecture and is addressed row-wise through a serially programmable (JTAG) sequencer. Discriminators have a common adjustable threshold with internal DAC. MIMOSA22 is the last chip (IDC-Intermediate Digital Chip), before the final sensor of the EUDET-JRA1 beam telescope, which will be installed on the 6 GeV electron beam line at DESY. In this paper, laboratory test results on analog and digital parts are presented. Test beam results, obtained with a 120 GeV pion beam at CERN, are also presented. In the last part of the paper, results on irradiated chips are given.

A Digital Monolithic Active Pixel Sensor Chip in a Quadruple-Well CIS Process for Tracking Applications

A CMOS sensor chip for charged particle detection has been developed and submitted for fabrication in a 0.18 μm Quadruple-Well (N&P-Wells, Deep N&P-Wells) CMOS Image Sensor (CIS) process. Improvement of the radiation hardness, the power dissipation and the readout speed of the mainstream CMOS sensors is expected with the exploration of this process. In order to ensure better charge collection and neutron tolerance, wafers with high-resistivity epitaxial layer have been chosen. In this paper a digital CMOS sensor prototype developed in order to validate the key analog blocks (from sensing element to 1-bit digital conversion) of a binary Monolithic Active Pixel Sensor (MAPS) in this process will be presented. The digital sensor prototype comprises four different sub-arrays of 20 μm pitch 64 × 32 pixels, 128 column-level auto-zeroed discriminators, a sequencer and an output digital multiplexer. Laboratory tests results including the charge-to-voltage conversion factor, the charge collection efficiency, the temporal noise and the fixed-pattern noise are presented in details. Some irradiation results will also be given.