Vorrichtung zur Beobachtung der dreidimensionalen Bewegung von Objekten, die in einer Flüssigkeit gehalten sind

Vorrichtung zur Beobachtung der dreidimensionalen Bewegung von Objekten (2), die in einer Flüssigkeit gehalten sind und deren Bewegung abhängig ist von den in der Flüssigkeit herrschenden örtlichen Eigenschaften wie der Temperatur, der chemischen Zusammensetzung oder den Lichtverhältnissen, wobei die Vorrichtung umfasst:- eine Beobachtungkammer (5) zur Aufnahme der mit den Objekten (2) versehenen Flüssigkeit,- eine erste Belichtungseinrichtung (6) mit einer Lichtquelle (7), eingerichtet zur lokalen Änderung der Eigenschaften in der Flüssigkeit,- eine optische Aufnahmeeinheit (12), eingerichtet zur optischen Erfassung der Objekte (2) in der Flüssigkeit, dadurch gekennzeichnet,dass die erste Belichtungseinrichtung (6) eingerichtet ist, gezielt eine Lichtmenge in einer gewünschten dreidimensionalen Verteilung in die Flüssigkeit der Beobachtungskammer (5) einzubringen, wobei die Flüssigkeit Bestandteile aufweist, die bei Einwirkung von optischen Lichtstrahlen einer bestimmten Wellenlänge ihre Eigenschaften ändern unddass die optische Aufnahmeeinheit eingerichtet ist, die Bewegung der Objekte (2) räumlich zu erfassen und in entsprechende elektronische Signale umzuwandeln

BIAS Against Asians, Black People, Hispanics/Latinos, Jews, and Muslims on Twitter

Hate speech on social media is a complex issue, challenging to quantify accurately. In an analysis of Twitter conversations (2020-2021) about Asians, Blacks, Jews, Hispanics/Latinos, and Muslims, 6,000 tweets were manually annotated by 4-6 students each. The agreement of annotators revealed 5-17% biased tweets, estimating 9.3 million biased tweets (majority agreement) and 6.1 million (75% agreement). Conversely, 19-54% of tweets called out bias against these groups, estimating 29.2 million tweets (majority agreement) and 17.8 million (75% agreement). Stereotypes varied across categories, portraying Asians as pandemic culprits, Black people as inferior and criminal, Jews as powerful with Holocaust denial, Hispanics/Latinos as illegal invaders, and Muslims collectively blamed for terrorism, often in discussions about Muslims in India

Simultaneous two-color imaging in digital holographic microscopy

We demonstrate the use of two-color digital holographic microscopy (DHM) for imaging microbiological subjects. The use of two wavelengths significantly reduces artifacts present in the reconstructed data, allowing us to image weakly scattering objects in close proximity to strongly-scattering objects. We demonstrate this by reconstructing the shape of the flagellum of a single-cell eukaryotic parasite Leishmania mexicana in close proximity to a more strongly-scattering cell body. Our approach also yields a reduction of approximately one third in the axial position uncertainty when tracking the motion of swimming cells at low magnification, which we demonstrate with a sample of Escherichia coli bacteria mixed with polystyrene beads. The two-wavelength system that we describe introduces minimal additional complexity into the optical system, and provides significant benefits

Cyclic Nucleotide-Specific Optogenetics Highlights Compartmentalization of the Sperm Flagellum into cAMP Microdomains.

Inside the female genital tract, mammalian sperm undergo a maturation process called capacitation, which primes the sperm to navigate across the oviduct and fertilize the egg. Sperm capacitation and motility are controlled by 3′,5′-cyclic adenosine monophosphate (cAMP). Here, we show that optogenetics, the control of cellular signaling by genetically encoded light-activated proteins, allows to manipulate cAMP dynamics in sperm flagella and, thereby, sperm capacitation and motility by light. To this end, we used sperm that express the light-activated phosphodiesterase LAPD or the photo-activated adenylate cyclase bPAC. The control of cAMP by LAPD or bPAC combined with pharmacological interventions provides spatiotemporal precision and allows to probe the physiological function of cAMP compartmentalization in mammalian sperm

Kinematics of flagellar swimming in Euglena gracilis: Helical trajectories and flagellar shapes

The flagellar swimming of euglenids, which are propelled by a single anterior flagellum, is characterized by a generalized helical motion. The 3D nature of this swimming motion, which lacks some of the symmetries enjoyed by more common model systems, and the complex flagellar beating shapes that power it make its quantitative description challenging. In this work, we provide a quantitative, 3D, highly resolved reconstruction of the swimming trajectories and flagellar shapes of specimens of Euglena gracilis. We achieved this task by using high-speed 2D image recordings taken with a conventional inverted microscope combined with a precise characterization of the helical motion of the cell body to lift the 2D data to 3D trajectories. The propulsion mechanism is discussed. Our results constitute a basis for future biophysical research on a relatively unexplored type of eukaryotic flagellar movement

Re-visiting the Protamine-2 locus: deletion, but not haploinsufficiency, renders male mice infertile

Protamines are arginine-rich DNA-binding proteins that replace histones in elongating spermatids. This leads to hypercondensation of chromatin and ensures physiological sperm morphology, thereby protecting DNA integrity. In mice and humans, two protamines, protamine-1 (Prm1) and protamine-2 (Prm2) are expressed in a species-specific ratio. In humans, alterations of this PRM1/PRM2 ratio is associated with subfertility. By applying CRISPR/Cas9 mediated gene-editing in oocytes, we established Prm2-deficient mice. Surprisingly, heterozygous males remained fertile with sperm displaying normal head morphology and motility. In Prm2-deficient sperm, however, DNA-hypercondensation and acrosome formation was severely impaired. Further, the sperm displayed severe membrane defects resulting in immotility. Thus, lack of Prm2 leads not only to impaired histone to protamine exchange and disturbed DNA-hypercondensation, but also to severe membrane defects resulting in immotility. Interestingly, previous attempts using a regular gene-targeting approach failed to establish Prm2-deficient mice. This was due to the fact that already chimeric animals generated with Prm2+/− ES cells were sterile. However, the Prm2-deficient mouse lines established here clearly demonstrate that mice tolerate loss of one Prm2 allele. As such they present an ideal model for further studies on protamine function and chromatin organization in murine sperm

Molecular mechanism underlying the action of zona-pellucida glycoproteins on mouse sperm

Mammalian oocytes are enveloped by the zona pellucida (ZP), an extracellular matrix of glycoproteins. In sperm, stimulation with ZP proteins evokes a rapid Ca2+ influx via the sperm-specific, pH-sensitive Ca2+ channel CatSper. However, the physiological role and molecular mechanisms underlying ZP-dependent activation of CatSper are unknown. Here, we delineate the sequence of ZP-signaling events in mouse sperm. We show that ZP proteins evoke a rapid intracellular pH i increase that rests predominantly on Na+/H+ exchange by NHA1 and requires cAMP synthesis by the soluble adenylyl cyclase sAC as well as a sufficiently negative membrane potential set by the spem-specific K+ channel Slo3. The alkaline-activated CatSper channel translates the ZP-induced pH i increase into a Ca2+ response. Our findings reveal the molecular components underlying ZP action on mouse sperm, opening up new avenues for understanding the basic principles of sperm function and, thereby, mammalian fertilization