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Kanazawa University research: Researchers define a protocol for narrow cantilever fabrication and high-resolution imaging of living cells using AFM

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KANAZAWA, Japan, Aug. 9, 2023 /PRNewswire/ — Researchers at Kanazawa University report in STAR Protocols procedural details and tips for nanoendoscopy-AFM, for capturing images of nanoscale structures inside living cells.

Images of nanoscale structures inside living cells are in increasing demand for insights into cellular structure and function they can reveal. So far, the tools for capturing such images have been limited in various ways, but researchers led by Takeshi Fukuma and Takehiko Ichikawa at Kanazawa University have now devised and reported a full protocol for using atomic force microscopy (AFM) to image inside living cells.

AFM was first developed in the 1980s and uses the changes in the forces between a sample surface and a nanoscale tip attached to a cantilever to “feel” surfaces and produce images of the topography with nanoscale resolution. The technique has grown increasingly sophisticated for extracting information about samples and at speeds sufficient for the tool to capture moving images of dynamics at the nanoscale. However, so far, it has been limited to surfaces. Other techniques exist that can provide a view of the inside of a cell but with limitations. For instance, there is electron microscopy, which is capable of resolving details at the nanoscale and smaller, but the required operating conditions are not compatible with living cells. Alternatively, fluorescence microscopy is regularly used on living cells, but while fluorescence techniques exist to increase the resolution, there are practical challenges that inhibit fluorescence imaging at the nanoscale. AFM suffers from neither limitation and by embellishing the tool with a nanoneedle to penetrate cells, Fukuma, Ichikawa and their collaborators have recently demonstrated the capability to image inside cells at the nanoscale, which they describe as nanoendoscopy-AFM.

In their protocol, the researchers break down the method for nanoendoscopy-AFM into 4 stages. The first few steps involve cell preparation and staining with a fluorescent dye and checking the fluorescence, which is used to identify the imaging area quickly. Next is the fabrication of the nanoneedles themselves, for which there are two options – either etching away a nanoneedle structure with a focused ion beam or building one up with electron beam deposition. Then comes the nanoendoscopy stage itself, and in the report, the researchers describe the approach for both 2D  and 3D nanoendoscopy. There are even details outlined to describe the best way to clean up after the nanoendoscopy images are captured before finally outlining the data processing needed to visualize the measured data. The method is replete with tips for successfully accomplishing each stage, as well as a guide for troubleshooting when things are not quite working out.

This technique should be suitable for the observation of intact intracellular structures, including mitochondria, focal adhesions, endoplasmic reticulum, lysosomes, Golgi apparatus, organelle connections, and liquid-liquid phase-separated structures. They conclude, “This protocol can be expected to become a standard tool for studying nanoscale structures.”

Figure: https://nanolsi.kanazawa-u.ac.jp/wp/wp-content/uploads/Graphical-abstract_EN.jpg

Caption: Overview of the method for observing actin fibers in living cells using nanoendoscopy-AFM.  © 2023 Ichikawa, et al., STAR Protocols

Glossary

Atomic force microscopy

Atomic force microscopy detects the attractive forces between a sample surface and tip from the resulting deflection of a cantilever that bears a nanoscale tip. Because the tip gradually tapers to such a small area, and the attractive forces dwindle so rapidly with distance, the topography of the surface can be obtained with nanoscale resolution. The technique was first reported shortly after the scanning conductance microscope, and although it was not the first to report an imaging resolution potential on the atomic scale, it had the great advantage of operation requirements that are compatible with non-conducting samples and living cells. It has also since provided valuable insights into dynamic insights with the development of a high-speed version of the tool at Kanazawa University. The development of this nanoendoscopy-AFM to penetrate cells extends the scope of the tool even further.

In one variation of AFM, the tip oscillates up and down as it is scanned over a structure, and the effect of changes in the resonance frequency of the oscillating tip gives an indication of the forces it is subject to. Tapping, dynamic, or AC AFM, as this has been described, has been attractive for imaging fragile structures as no contact is made. Here it is used inside the cell for 2D nanoendoscopy. An alternative is 3D nanoendoscopy, where at each point that is raster scanned, the tip is retracted by a specified distance, which may be between 0.1 and 1 micrometres. The measured force until it reaches the set point is then noted at each point.

Reference

Takehiko Ichikawa, Mohammad Shahidul Alam, Marcos Penedo, Kyosuke Matsumoto, Sou Fujita, Keisuke Miyazawa, Hirotoshi Furusho, Kazuki Miyata, Chikashi Nakamura and Takeshi Fukuma

Protocol for live imaging of intracellular nanoscale structures using atomic force microscopy with nanoneedle probes, STAR Protocols September 15, 2023.

 DOI:10.1016/j.xpro.2023.102468

URL: https://doi.org/10.1016/j.xpro.2023.102468

Contact
Hiroe Yoneda
Senior Specialist in Project Planning and Outreach
NanoLSI Administrative Office
WPI Nano Life Science Institute (WPI-NanoLSI)
Kanazawa University
Kakuma-machi, Kanazawa 920-1192, Japan
Email: [email protected]
Tel: +81 (76) 234-4550

About Nano Life Science Institute (WPI-NanoLSI)

https://nanolsi.kanazawa-u.ac.jp/en/ 

Nano Life Science Institute (NanoLSI), Kanazawa University is a research center established in 2017 as part of the World Premier International Research Center Initiative of the Ministry of Education, Culture, Sports, Science and Technology. The objective of this initiative is to form world-tier research centers. NanoLSI combines the foremost knowledge of bio-scanning probe microscopy to establish ‘nano-endoscopic techniques’ to directly image, analyze, and manipulate biomolecules for insights into mechanisms governing life phenomena such as diseases.

About Kanazawa University

http://www.kanazawa-u.ac.jp/e/ 

As the leading comprehensive university on the Sea of Japan coast, Kanazawa University has contributed greatly to higher education and academic research in Japan since it was founded in 1949. The University has three colleges and 17 schools offering courses in subjects that include medicine, computer engineering, and humanities.

The University is located on the coast of the Sea of Japan in Kanazawa – a city rich in history and culture. The city of Kanazawa has a highly respected intellectual profile since the time of the fiefdom (1598-1867). Kanazawa University is divided into two main campuses: Kakuma and Takaramachi for its approximately 10,200 students including 600 from overseas.

 

View original content:https://www.prnewswire.co.uk/news-releases/kanazawa-university-research-researchers-define-a-protocol-for-narrow-cantilever-fabrication-and-high-resolution-imaging-of-living-cells-using-afm-301896615.html

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Blockchain Press Releases

Bitrue Gears Up for 2024 Bitcoin Halving with Trading Competition

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SINGAPORE, April 12, 2024 /PRNewswire/ — Leading cryptocurrency exchange Bitrue unveiled a new competition available to all users on the platform that comes with a total prize up to $850,000 USDT in a recent X announcement.

The contest is being held in honor of the much anticipated Bitcoin Halving – an event that occurs once every four years to regulate the supply of new BTC entering the digital asset economy, and helps ensure long-term value through scarcity of supply.

The competition will rank traders according to their trading volume on the exchange during the contest period. The competition begins on Thursday April 11th and will continue until the halving event occurs (estimated April 20th). The prizepool will increase according to the number of valid participants in the competition, with the grand prize winner set to take home 25% of the total reward (212,500 USDT). All participating users will have a chance to win prizes regardless of their final rank.

With all eyes on this historic event, Bitrue is thrilled to be able to offer an exciting and rewarding way for its 10 million users to mark the occasion and provide them with an opportunity to earn a life changing amount of money. Interested parties should sign up on the Bitrue exchange now in order to be ready for when the contest goes live on April 11th.

About Bitrue

Launched in July 2018, Bitrue is a diversified digital exchange that supports trading, loans and investments. Bitrue aims to utilize blockchain technology to bring financial opportunities to everybody regardless of their location or financial position. It has offices in Asia and Europe, and continues to develop new features at a rapid speed to fully service the new wave of the digital economy. More information is available at Bitrue’s website.

Logo – https://mma.prnewswire.com/media/2385358/Bitrue_BitrueOfficial_Bitrue_Logo.jpg 

Cision View original content:https://www.prnewswire.co.uk/news-releases/bitrue-gears-up-for-2024-bitcoin-halving-with-trading-competition-302115311.html

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Financial industry bodies defend permissionless blockchains against Basel Committee’s classification

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Five financial industry bodies have pushed back against the treatment of permissionless blockchains by a global banking supervision authority.

In December, the Basel Committee on Banking Supervision (BCBS) published a report on proposed amendments to bank capital requirements for digital assets, stablecoins, and tokenized assets.

The report classified all permissionless blockchains as high-risk, claiming that some risks could not be mitigated through existing solutions. BCBS was particularly concerned about banks’ lack of control over third parties who conduct most operations on these blockchains. It also warned about their privacy, finality, liquidity, and political, legal, and policy risks.

In response, five global financial industry regulators have defended permissionless blockchains. In a joint response, they stated that the industry “has all necessary expertise and robust compliance frameworks to fully identify, manage and mitigate these risks.”

The five are the International Swaps and Derivatives Association, the Global Financial Markets Association, the Institute of International Finance, the Futures Industry Association, and the Financial Services Forum.

Blockchain’s application in the financial industry is evolving, and regulators must not disincentivize banks from exploring the technology, the regulators stated. By putting up unnecessary hurdles, the BCBS would only push these institutions to the non-regulated shadow banking space, which would be riskier for them.

The regulators further noted that dozens of global banks have conducted successful pilots using permissionless blockchains. These pilots have shed more light on the technology’s application and allowed them to understand and control emergent risks.

The BCBS approach is unfair to blockchain and veers away from the regulator’s long-held “same asset, same risk” approach, they added.

“While we acknowledge that risk mitigation techniques are evolving for permissionless crypto assets…we are confident that solutions already exist in respect of specific use cases,” the five stated.

They believe deciding whether to build on permissionless blockchains should be left to the banks.

The financial sector has been a leader in blockchain adoption, with some, like JPMorgan (NASDAQ: JPM), developing their own permissioned networks, albeit unsuccessfully. However, most have relied on existing solutions to build applications spanning settlement, bond issuance, tokenization, etc.

Source: coingeek.com

The post Financial industry bodies defend permissionless blockchains against Basel Committee’s classification appeared first on HIPTHER Alerts.

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Partnership between Siren and Chainalysis to trace blockchain transactions and disrupt illicit activities

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Siren, the all-in-one investigation platform company on a mission to keep people, assets and networks safe, and Chainalysis, the blockchain intelligence platform, announced details of a strategic partnership at the annual Chainalysis Links event in New York.

As cryptocurrency becomes more mainstream and used by both good and bad actors, the partnership addresses the increasing need for more sophisticated intelligence technology to fight illicit cryptocurrency activity.

Chainalysis’ mission
Many believe cryptocurrency is the future, and it is expected that blockchains will soon be the world’s primary mechanism for the exchange of value. Chainalysis’ mission is to build trust in blockchains, setting the stage for the mass adoption of crypto in a way that gives participants safety and security.

Its data powers investigation, compliance, and market intelligence software that has been used to solve some of the world’s most high-profile criminal cases and grow consumer access to cryptocurrency safely.

AI-driven search functionality

Siren is an AI-driven investigative intelligence platform that is used by some of the world’s pioneering Law Enforcement, National Security and Cyber threat investigators. Siren connects and enriches local classified information with external vendor data as well as data available from public sources to identify patterns, actors and events.

Siren’s AI-driven search functionality dramatically reduces the complexity in searching for intelligence-like browsing the internet or shopping online anyone can search for intelligence links, regardless of technical abilities, across multiple data sources and get immediate results on both desktop and mobile devices. Using patented technology, Siren delivers a modern search experience at enterprise speed and scale, making advanced intelligence queries accessible to all across the organisation.

Chainalysis Investigations solution

Through the collaboration, illicit activity is easier to identify in real-time across multiple data sources, including Chainalysis’ on-chain data, allowing investigators and analysts to map addresses to named services, detect illicit activity as it happens, and build cases.

The Siren platform leverages the Chainalysis Investigations API (IAPI), part of the Chainalysis Investigations solution. The Investigations API is a REST-based API that programmatically helps to prioritise and accelerate investigations. This ensures investigation teams can focus on the most important cases with the highest likelihood of success. The IAPI enables data engineers and scientists to enrich existing datasets with Chainalysis insights to enhance large-scale investigations, surface new leads, and uncover previously unknown connections between different cases for deconfliction.

New York at Links 2024
John Randles, CEO of Siren, said: “Linking to crypto across multiple data sources is pivotal for investigators. We’re delighted to announce the partnership here in New York at Links 2024. We were lucky to attend a couple of Chainalysis events last year and experience first-hand how the synergies in our offerings and missions.”

John Randles will speak on Day Two of Chainalysis Links 2024 on the Intersection of AI and Crypto panel. Shannon Hughes, Senior Director, Head of Business Development and Partnerships commented: “Cryptocurrency is no longer just a niche specialty for elite cyber law enforcement units – our customers find connections to crypto across national security, crime, fraud, and cyber threats. This partnership enables investigators to combine Chainalysis data with other data sources to identify intelligence signals and actionable leads across specialties. This is the future of harnessing the transparency of blockchains to build a safer economic system.”

Source: sourcesecurity.com

The post Partnership between Siren and Chainalysis to trace blockchain transactions and disrupt illicit activities appeared first on HIPTHER Alerts.

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