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Legal Question in Landlord & Tenant Law in Illinois
I would like to know if I can deduct the lost rent I incurred when my subletter broke the lease agreement and I had to find a new roommate on a moment's notice? It was a month to month agreement and I gave her 30 days notice on July 1st to be out by August 1st. She chose to move out that weekend. I was unable to fill her room until July 14th. She broke the lease by giving her key to the house to a friend that we had never met, never announced him nor approved his stay, and he walked in our home unaccompanied in the middle of the night. She was no where to be found or reached. I almost called the cops because I thought he was an intruder. He was made to leave and leave the key. I gave her notice the very next day. We had an oral agreement about no guests unaccompanied or unannounced, and I have texts that also have this agreement established.
1 Answer from Attorneys
Without reading the entirety of your email, which may require additional information to be able to discuss properly, your basic question is whether a subtenant can be held liable for damages you incur as tenant when the subtenant breaches the sublease leaving you with no rental income to continue to pay your obligation under your main lease. The answer is yes, in general. Proving it, and dealing with the costs of a lawsuit (and attorney fees, if any), may or may not be cost effective.
Related Questions & Answers
How long do I have to get out of a lease after it is signed for an apartment? Asked 11/10/13, 10:16 am in United States Illinois Landlord & Tenants
My landlords tee broke our car windshield Is he liable Asked 10/22/13, 1:23 pm in United States Illinois Landlord & Tenants


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It was the average day in the Mansion, you were avoiding any unpleasant encounters. You were fed up with being food. You lay there in your bed wondering how to avoid them completely without causing trouble. Could you hide? No, they will easily find you. Could you be busy doing something and tell them no? Probably not. Could you go back to sleep? Definitely not. You really had no options whatsoever. How troublesome. You got out of bed, your [Hair Length] [Hair Color] locks nicely placing on your form. "Ehh?! I feel exhausted and I haven't even done anything.." You muttered to yourself. You then let out a small sigh of annoyance. Same events, different day… You heard a gentle sound of breathing from your bed. As you looked over you noticed one of the more tolerable ones, Shu. You didn't even notice him there. You blinked a few times staring at him. "When did he get there?" You whispered, trying not to wake him. "Too noisy." He muttered opening one eye, glaring at you. "S-Sorry.." You whisper as you got your clothes to get ready. "Shu, not to be rude but can you leave for a moment?" Of course, he didn't. You really wanted to change and he was being plain bothersome. "Shu. I demand you leave." You raised your voice. "I refuse." He smirked. "Then go back to sleep!" He yelled. "Trying. But you're way too noisy." You let out a frustrated growl and stormed out of there, heading to the bathroom.
Brushing your [Hair Color] hair, you noticed a reflection in the mirror. "Ayato, not now." You muttered continuing to brush your hair. "That's no way for food to talk." He whispered in your ear. You slapped him away, getting really annoyed. "Ayato, I'm not in the mood, please go away." you raised your tone, hoping he'd leave. "Acting tough makes me want to stay, heh." Ayato made a devilish smirk, that monster. You sighed, putting on your makeup. Simple makeup, nothing too fancy. "Why do you put that stuff on? It's gross." He cringed. Just then, an idea popped into your head. "Oh, really now?" You smirked, putting makeup over your neck and a bit on your wrists. "Why you--" He was cut off. "There all done!" You gave Ayato an evil smirk. "A monster.." He muttered. Just then, to settle it, you also put perfume almost all over your body. Now, your skin must smell and taste horrible. "Haters gonna hate." You grinned at him, walking to go get your socks. "W-Witch." He spoke, walking out of the bathroom. You smiled, feeling quite proud of your accomplishment. Often, you would put a few pairs thigh high socks on, in case of Raito. Which is what you did today. You've been doing the best you could to not get bitten and they definitely don't bug you AS much. Giving yourself the final scan, making sure you looked decent, you walked out of the bathroom. "Waking up, check, get ready, check." You muttered, sighing right after.
You happened to be in the garden in the front yard. This was your day of relaxing and the Sakamaki boys are definitely not going to ruin it. You grinned finding one pinkish rose. You then decided to pick it, hoping no one would notice. Not that many of them care about petty flowers. As you made your way through the garden, once again you spotted Shu. He was sleeping on the bench with earbuds in each of his ears.Does he sleep EVERYWHERE? "Shu…?" You began as he opened his eyes. "Won't you catch a cold being out here so long? It is in fact, a nice day, but, it's a bit chilly…" He stared at you, blinking a few times. "Then warm me up…" He lazily spoke. Your [Eye Color] orbs widened at his request. He's got to be kidding. You sighed and wrapped your arms around him. You remained like that for a while. As you watched Shu sleep, you gently pushed his bangs out of his face. You let out a small sigh. "Now I'm getting cold.." You muttered. Just then, large arms wrapped around you as you felt breathing on your neck. "Then why didn't you say so?" He whispered. "Because you would have called me noisy." You smirked at him as he grinned. "Shut up.." He blushed whilst giving you a smirk as well. As he went to go bite you, he sniffed your neck and cringed. "That's overkill isn't it?" He gave you a glare. "Saved me from Ayato." You closed your eyes for a moment before opening them. "It was necessary." You spoke in a serious tone. "Whatever." He replied, getting up and heading inside.
You sat there a little bit longer. 'Did I hurt his feelings or something?!' you thought. Whatever it was, he seemed upset. That's when you decided, you'd go after him to figure him out. You wanted to understand what was going through his head. But first…how bout a shower, this perfume was bugging you as well. You got up from the bench heading inside, thoroughly washing there areas with the perfume. You sneezed. 'Bless me…' you thought to yourself. You sighed, you put way too much perfume on. Oh well, that the most you could get off. Getting out of the shower, you wrapped a towel around you and headed to your room. The door was locked. "What the hell…" You cursed under your breath. You knocked. Knock…Knock…Knock…No answer. "Shu? Are you in there?" You got ready to ram the door when the door opened, making you run into Shu. He's like a boulder, he barely moved an inch. "Too noisy.." He closed his eyes out of annoyance. As he walked out. Finally you can get dressed in your room for once. You had to go to sleep soon so you put on your pyjamas a little bit earlier than usual. Now time to go find Shu.
As you set out to find Shu, you noticed him sleeping in the bath…again. "Shu! You're getting your clothes soaked!" You yelled. "I was too lazy to take them off…" He muttered under his breath. "Too bothersome." You said, imitating him. "…" Shu was silent after that remark. He finally broke the silence. "[Your Name], come here." He whispered softly. You did as asked. "I'm here, what is it?" You knelt down to his current level. Suddenly, he pulled you into a deep embrace. "Aishitaru…(Translation: I love you…)" Your [Eye Color] orbs widened. Soon, you melted into his embraced. "Aishitaru, Shu." He got up, picking you up bridal style. "Eh?! Where are we going?!" he began walking to your bedroom. Opening the door, he slowly closed it once you entered. Tossing you on the bed, he opened his mouth to speak. "Go to sleep…" He whispered. "Go back to sleep." You replied as you patted the spot beside you. Shu did as asked and fell asleep within 10 seconds of impact. "That was fast…" Your face as astonished at how much he can sleep. You laid down, hugging him. Thusly, you fell asleep with the most gentle vampire you know.

subset: commoncrawl


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Does anyone know of a place in NYC/tri-state area that sells a good Damascus-style falafel sandwich? By Damascus-style I mean: thin Lebnani bread, rolled up with fresh tomato, cucumber, thin pickled vegetables, fresh mint, sweet bbq/pomegranate sauce, tahini, hot sauce, etc. The closest I've had is Aloosh in Paterson, NJ which is sadly now closed.
The falafel I have had in New York is almost always in a round puffy pita, with lettuce or cabbage, hummus, diced tomatoes, etc. and is just a different beast.
Many thanks.


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Campus News 2006
For Immediate Relase
August 31, 2006
Unabomber's Brother To Discuss Death Penalty at Schreiner University Sept 12
Schreiner University is bringing the brother of the infamous Unabomber Ted Kaczynski to campus on Tuesday, September 12 for a lecture entitled "Confronting Violence, Seeking Justice — The Death Penalty in America: A Conversation with David Kaczynski." This event, sponsored by the Schreiner Student Activities Board, will take place at 7 p.m. in the ballroom of the Floyd & Kathleen Cailloux Campus Activity Center on the Schreiner campus. It is free and open to the public.
The Kaczynski family became a focus of media attention in April 1996 with the arrest of Ted Kaczynski, who the FBI had code-named The Unabomber. Kaczynski was responsible for a 17-year reign of terror involving mail bombs that killed three people and injured 23 others. Soon after Kaczynski's arrest, it was leaked that Kaczynski's brother David, a youth counselor, and David's wife Linda Patrik, a college professor, had approached the FBI with their suspicions about Kaczynski, resulting in his arrest.
The couple soon felt betrayed, first when the U.S. Justice Department broke its promise to keep the family's role in the investigation confidential, and again when it sought the death penalty despite Kaczynski's serious mental illness. David and Linda fought for nearly two years to prevent the execution of David's brother, and ultimately failed in their quest for justice.
David Kaczynski has spoken widely and appeared on national television shows voicing his opposition to the death penalty. In 1999, David and Linda were honored by the New York State Bar Association with its annual Justice Award – the organization's only presentation to non-lawyers. David has addressed community, school, and professional audiences on a range of topics, including the death penalty, mental illness, non-violence, and ethical decision-making. In July 2001, after a nationwide search, David was selected as the new Executive Director of New Yorkers Against the Death Penalty – a statewide organization that unconditionally opposes capital punishment.
For more information on this and other upcoming events, please contact Jennifer M. Hudson-Velazquez at email@example.com or call 830-792-7283.
To have an e-mail reminder sent to you prior to an event please visit our CAMPUS CALENDAR. Just click on the event you are interested in and choose the option "set reminder" on the detail page.


text: Video News: Canada On This Day - June 16, A Shot Of History Get On This Day delivered free to your inbox: http://bit.ly/14agw7n http://www.worldcrunch.com 1846 – Pope Pius IX is elected Pope beginning the longest reign in the history of the papacy. http://en.wikipedia.org/wiki/Pope_Pius_IX https://www.youtube.com/watch?v=8uyjStbUh0I 1940 – World War II: Marshal Henri Philippe Pétain becomes Chief of State of Vichy France (Chef de l'État Français). http://en.wikipedia.org/wiki/Philippe_P%C3%A9tain https://www.youtube.com/watch?v=_z4SUYs14X0 1976 – Soweto uprising: a non-violent march by 15,000 students in Soweto, South Africa turns into days of rioting when police open fire on the crowd. http://en.wikipedia.org/wiki/Soweto_uprising https://www.youtube.com/watch?v=wCW3DqzopsM 1978 – Daniel Brühl, Spanish-German actor (Good Bye, Lenin!) http://en.wikipedia.org/wiki/Daniel_Br%C3%BChl https://www.youtube.com/watch?v=Kehu8QBHCCk Produced by Meiqi AN © DailyMotion - Friday, June 5, 2015


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NSF International, The Public Health and Safety Company, has opened the new FreshCheck laboratory at the NSF International World Headquarters in Ann Arbor, MI. NSF FreshCheck is a comprehensive microbial monitoring and auditing program for grocery stores and supermarkets.
The transition from St. Paul, MN to the new laboratory facility was completed in just three months after the acquisition of FreshCheck. The new FreshCheck laboratory in Ann Arbor utilizes state-of-the-art analytical equipment and timely web-based reporting to provide customers with quick turnaround time and improved customer service.
The NSF FreshCheck Food Safety Risk Management Program includes three components: 1) audits to evaluate food safety, sanitation and customer perceptions; 2) microbial monitoring to test for bacteria that may cause illness, spoilage and poor quality in perishables; and 3) FastCheck™, an immediate response to customer foodborne illness allegations, which includes testing of suspect products. The new FreshCheck laboratory allows all of these services to be performed in one central location.
"Having so many programs and services offered at one location is a great benefit. The new laboratory, which is an integral part of their large campus-like facility, provides us with all the food safety resources and expertise we need," said Orhan Uner, Director of Central Procurement, Nash Finch Company. "In addition to rapid microbiological testing, we also have access to the NSF Center for Public Health Education."
"We are very pleased with the new FreshCheck laboratory. The larger, highly advanced facility and the increased number of staff allows for unparalleled customer service," said Nancy Culotta, Vice President, Retail Food Safety Programs. "Customers may now take advantage of all the food safety services that NSF offers including food processor audits, education and consulting services."
NSF FreshCheck, a leader in food microbiological testing and sanitation audits, provides a broad range of comprehensive food safety services. NSF International, a not-for-profit, non-governmental organization, is the leading global provider of public health and safety risk management solutions. NSF provides product certification and safety audits for the food and water industries. Other services include management systems registrations delivered through NSF International Strategic Registrations, Ltd., and education through the NSF Center for Public Health Education. NSF is a World Health Organization Collaborating Centre for Food Safety and for Drinking Water Safety and Treatment. Serving companies in 88 countries, NSF was founded in 1944 and is headquartered in Ann Arbor, MI USA.


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You Need To Keep Your Kids Safe
"Bankrupt At Birth" is the title of the book that I'll be giving away later this afternoon (Thursday) on my Puffman Musical Trivia. "Bankrupt At Birth" explains why child ID theft is on the rise & how it's happening under parent's noses. The NFL's Steve Young says "our children are now living in an age where social media is central to their lives. You need to read this book to understand the dangers they face & what YOU need to do to keep them safe". Joe Mason, the general manager of IDENTITYGUARD.COM & a senior VP at Intersections Inc is the author along with Steve Schwartz. Mr. Mason has played a role in protecting the identities of over 3 million Americans & has provided his expert advice on CNN, Fox, & ABC. The CEO of Children's Miracle Network Hospitals, John Lauck says "this book shows you child identity theft is a very big deal, & teaches you what you need to do about it". Visit www.BankruptAtBirthBook.com for additional resources & be the first caller with the correct answer to my Pufmman Musical Trivia question this afternoon at 4:30 & win yourself a copy of this timely read. By the way, the infamous Mary Lou Retton, Olympic Champion & Children's Miracle Network Hospitals Board Member has written the forward & 100% of the book's profits are going to the Children's Miracle Network Hospitals! Certainly a most timely book in this day & age.


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There is no single approach to teaching that works best. What we do in the classroom depends on who we are, who our students are, what we are teaching, and many other factors. There are some guiding principles that can help use to create the condition that support and promote learning.
Looking at this graphic, it seems that active classroom activities may help students learn and retain information in ways that traditional lectures do not. Get students engaged and excited about their learning using Active Learning methods.
Look for ways to prompt students to think more deeply about course concepts and the learning process through Prompts That Get Students to Analyze, Reflect, Relate, and Question.
The Flipped Classroom
The Flipped Classroom is a teaching approach that moves lectures outside of the classroom to allow classroom time to be spent in more dynamic learning activities. Before arriving to class, students often watch recorded video lectures and read through lecture materials, complete a basic assessment to ensure they have understood basic concepts. They then arrive in the classroom ready to engage with their peers in discussions and other student-centered learning activities. Looking at the Learning Pyramid provides some incentive to move away from lectures to more practice-based activities in the classroom.
Overview: Watch one instructors approach in her YouTube video
The Flipped Classroom: Pro and Cons
Flipping A Class: A How-to Guide for Beginners
This teaching strategy seems to be gaining momentum because it gets results. Shaking up tried and true teaching methods can be controversial and this approach is not for everyone, but for some teachers and students, it is breathing new life into the learning experience. Read more from the NYTimes on the Flipped Classroom.


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table of contents
Bone morphogenetic and osteogenic proteins (BMPs/OPs) are soluble mediators of tissue …
Home » Biology Articles » Bioengineering » Soluble, insoluble and geometric signals sculpt the architecture of mineralized tissues » Implications for bioengineering
Implications for bioengineering
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More than 34,000 students head back to school this week in Minneapolis and the excessive heat will make it uncomfortable for some.
Minneapolis Public Schools say they will provide emergency water on request to schools that are not fully air-conditioned. The water will be offered to students and staff who don't have easy access to drinking fountains.
Eighteen of the district's buildings do not have air conditioning. Another 11 have limited air conditioning.
In southwestern Minnesota classes are already underway in some districts.
John Landgaard, superintendent of the Worthington School District, said all of his buildings have air-conditioning.
"Our kitchen in the high school is not air conditioned, so we have to pay attention to our employee needs and make sure they get out and get fresh air, so they don't get overheated," he said.
Excessive heat is expected to blanket much of the region through Wednesday.


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WA still making up its mind on federal ed waivers
Washington state education officials haven'treally decided yet if the state will apply for a waiver...
Long sentence sought in Wenatchee slaying
The Chelan County prosecutor says he'llseek an exceptional sentence against a man accused of...
Pasco Fire Started by 9-Year Old
Investigators say a fire that damaged abathroom at Pasco's Markham Elementary School was started by...
Fake Baby Bump - Movie Deal
Faking her pregnancy is the high school projectthat keeps paying off for one Toppenish teenager....
Finance Minister resigns over protester deaths in Cairo
Egyptian minister resigns over protester deaths
Egyptian Coptic Christians feel like second class citizens
Christians under siege in post-revolution Egypt
Category: General News
Idaho Urban Renewal Bill Dies
BOISE, Idaho (AP) — A bill giving Idaho voters more power over urban renewal projects has died in a state House committee after two hours of testimony. The legislation rejected yesterday would have given citizens a say over certain land redevelopment plans, a power currently reserved for city councils and zoning commissions. Some said the measure would stretch out the timeline for approving new projects.


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E.V. Peterson: 'Irresponsible' city
January 24, 2011
To the editor:
This is regarding Craig, the city of no sidewalks.
I, like Ms. Rohrich, am gravely concerned that our school children have to walk to school on the streets.
I observe, on a daily basis, children, joggers, walkers, etc., struggling to use streets that have no sidewalks to get around on.
I have lived in many communities, some smaller, some larger than Craig. All of them have sidewalk ordinances, except Craig.
Why is there no concern for the safety of our pedestrians?
Recommended Stories For You
Why are there no sidewalks required in this city?
It is a travesty that there is such a lack of concern for the safety of our walking residents.
One of these days, a child is going to be killed and I hope the parents sue the pants off of, not only the driver, but the irresponsible city of Craig!
E.V. Peterson
Click here to have the print version of the Craig Daily Press delivered to your home.


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LIght Emitting Diode (LEDs) Position Update
Light Emitting Diodes (LEDs) are devices that licensed estheticians should be allowed to use for beautifying & stimulating the skin. When used in accordance with manufacturer instructions they are safe, and do not damage the skin. The range of most LED devices for the intended use of beautifying & stimulating the skin are 410 nm – 945 nm. (nm = nanometers)
LED devices are not lasers.
The light particles emitted from an LED device are scattered or diffused, which means the light is dispersed in many different directions. Laser beams are focused or concentrated. A good analogy would be that of a garden hose. The hose on "wide spray setting" is like that of a LED, and the hose's "stream setting" would be the beam of a laser.
Use of LEDs, does not require the same safety precautions.
Light emitting diodes are different from laser diodes and are not subject to the Federal laser product performance standard.(1)
LED safety history.
Associated Skin Care Professionals (ASCP) insures over 13,500 licensed skin care professionals in the United States. There has never been a claim or incident reported as a result of the use of any LED device or in conjunction with the use of any LED device.
Marine Agency Corp is a leading provider of professional liability insurance to estheticians and day spas throughout the United States. Our policies have included coverage for LED (light emitting diode) services for at least ten years. To date, I am not aware of any professional liability claims for these services.
Professional Program Insurance Brokerage insures hundreds of estheticians in the United States. The use of LEDs by their policyholders has proven—by its track record—that these devices have been used safely and effectively by licensed estheticians in the United States.
- About NCEA
- Member Center
- Political Advocacy
- Esthetic Teachers Esthetic Schools
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Artist Bryan Zanisnik was half an hour into reading Philip Roth's The Great American Novel inside a Plexiglass case surrounded by junk when the author's lawyers served him with a cease and desist letter. Mid-performance. Opening night. The kicker? He was reading it silently.
The Abrons Art Center on the Lower East Side hosting Zanisnik's Every Inch a Man are "shocked" but "not very concerned," they tell ArtNet. "The grounds of the cease and desist are erroneous. We're not reading anything out loud or reproducing anything — there's no chance here of copyright infringement." Is this the new age of Thought Copyright Police or did the lawyers just make an oops? Why's the Abrons Art Center site down? Uh-oh.


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We would like to wish Mazal Tov to Ushi and Chani (nee Bukiet) Steinmetz (Brooklyn, NY) on the birth of their son.
May the newborn be a source of Nachas to his parents, grandparents, relatives and Klal Yisroel.
To submit a Simcha or Mazal Tov please email us at email@example.com!


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When I get ideas for food, it kind of just hits me and then, since he will be the taster, I always run it by Ryan too. Well, when I said to him "I'm going to make a watermelon salad with feta cheese, onions and mint, " his response was, and I quote, "Well that doesn't sound good at all." Not one to be discouraged, I made it anyway. And guess what….Ryan devoured it.
I too thought the idea of watermelon, cheese and onions was a crazy combination but when I looked in my copy of The Flavor Bible and noticed that it said feta was a great combination with watermelon I was a intrigued. And alas, the book did not let me down. It was fantastic. It was cool. It was flavorful. The watermelon sweet and the cheese creamy. There is no other word for it, it is simply divine.
Combine all ingredients together in a bowl. Cover and refrigerate for 30 minutes. Serve.


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Fabrication of Si heterojunction solar cells using P-doped Si nanocrystals embedded in SiN x films as emitters
- Ping-Jung Wu1,
- Yu-Cian Wang1 and
- I-Chen Chen1Email author
© Wu et al.; licensee Springer. 2013
Received: 2 June 2013
Accepted: 26 October 2013
Published: 5 November 2013
Si heterojunction solar cells were fabricated on p-type single-crystal Si (sc-Si) substrates using phosphorus-doped Si nanocrystals (Si-NCs) embedded in SiN x (Si-NCs/SiN x ) films as emitters. The Si-NCs were formed by post-annealing of silicon-rich silicon nitride films deposited by electron cyclotron resonance chemical vapor deposition. We investigate the influence of the N/Si ratio in the Si-NCs/SiN x films on their electrical and optical properties, as well as the photovoltaic properties of the fabricated heterojunction devices. Increasing the nitrogen content enhances the optical gap E04 while deteriorating the electrical conductivity of the Si-NCs/SiN x film, leading to an increased short-circuit current density and a decreased fill factor of the heterojunction device. These trends could be interpreted by a bi-phase model which describes the Si-NCs/SiN x film as a mixture of a high-transparency SiN x phase and a low-resistivity Si-NC phase. A preliminary efficiency of 8.6% is achieved for the Si-NCs/sc-Si heterojunction solar cell.
Materials consisting of silicon nanocrystals (Si-NCs) embedded in a dielectric matrix are one promising candidate to realize Si-based third-generation photovoltaic devices owing to their potential benefits of utilizing the visible light of terrestrial solar spectrum and overcoming the efficiency limit of crystalline Si (c-Si) solar cells [1–5]. Sub-stoichiometric Si-based dielectric materials, such as SiO x , SiN x , and SiC x , have been investigated for synthesis of Si-NCs [6–11]. The formation of Si-NCs is based on phase segregation and crystallization in Si-rich dielectric films during the post-annealing process [12].
The low conductivity of Si-NCs embedded in dielectric films limits their applications for the manufacturing of optoelectronic devices. For this reason, impurity doping in Si-NCs embedded in SiO2 has been demonstrated to modify the electrical properties of the layers, although there is some debate about the feasibility of doping in Si-NCs [13, 14]. In addition to impurity doping, the choice of the surrounding dielectric matrix also plays a crucial role in charge carrier transport. Although the formation of Si-NCs in the SiO2 matrix has been investigated in detail [12, 15], the carrier transport ability in the Si-NC network is generally insufficient due to the large energy barrier of the surrounding oxide matrix. Charge carrier transport through narrower bandgap dielectrics, such as Si3N4 or SiC, seems to be more feasible. Compared with SiO2 and SiC, Si3N4 may offer a compromise as a dielectric matrix for the Si-NC network used in solar cell applications since it possesses a medium bandgap (approximately 5.3 eV) which could reduce the energy barrier for carrier transport and also effectively avoid parasitic absorption. However, doped Si-NCs embedded in a SiN x matrix (Si-NCs/SiN x ) have not received much attention.
In this work, we present initial fabrication and characterization results of Si heterojunction solar cells using P-doped Si-NCs/SiN x films as emitters. The P-doped Si-NCs/SiN x films were prepared by electron cyclotron resonance chemical vapor deposition (ECRCVD) followed by high-temperature annealing, and the influence of the chemical composition (N/Si ratio) on their physical properties was investigated. The photovoltaic properties of the fabricated heterojunction devices were also examined as a function of the N/Si composition ratio in the P-doped Si-NCs/SiN x films.
Fifty-nanometer-thick, homogeneous Si-rich silicon nitride (SRN) films containing phosphorus were deposited by a homemade ECRCVD system on single-side polished p-type (100) single crystalline Si (sc-Si) substrates with a thickness of 675 μm and a resistivity in the range of 5 to 20 Ω cm. Before placing into the deposition chamber, Si substrates were cleaned with acetone and rinsed in deionized water followed by removal of native oxide on Si wafers using a diluted HF dip (5%). The mixed SiH4, N2, Ar, and PH3 gases were then introduced into the deposition chamber at 10 mTorr for film growth. The applied microwave power and the substrate temperature were kept on 700 W and 200°C, respectively. In order to study the influence of the Si/N ratio on film properties, both SiH4 and PH3 flow rates were kept constant during film growth, while the gas mix ratio (Rc) defined as N2/SiH4 was varied in the range 0.73 ≤ Rc ≤ 0.83. The formation of Si-NCs in as-deposited SRN films was carried out by post-growth annealing in a quartz tube furnace at 950°C in N2 ambient. Samples with a 1 cm × 1 cm area were used for subsequent fabrication of heterojunction solar cells. Aluminum films deposited by electron gun evaporation were used as contact electrode layers in the solar cells. The front contact on top of the Si-NCs/SiN x film was defined by a shadow mask during Al deposition, while the rear contact covered the full back area of the cell. After metallization, the samples were heated at 500°C for 3 min to improve the electrical properties of the contacts.
For the characterization, the bonding configurations of the Si-NCs/SiN x films were identified by X-ray photoelectron spectroscopy (XPS). Micro-Raman spectroscopy and transmission electron microscopy (TEM) were used to investigate the crystallization behavior in SRN films after post-growth annealing. Fused quartz wafers were used as substrates for Raman studies to avoid the signal contribution from Si substrates during Raman measurements. X-ray diffraction (XRD) was used to evaluate the Si-NC size of annealed samples. The photovoltaic properties of the fabricated heterojunction solar cells were evaluated at room temperature based on the illuminated current density versus voltage (J-V) and the internal quantum efficiency (IQE) characteristics under 1-sun air mass 1.5 global spectrum.
Results and discussion
From Figure 6, the Jsc is increased from 21.3 to 28.2 mA/cm2 with increasing Rc value. This trend could be ascribed to the lower parasitic absorption in the Si-NCs/SiN x film with a higher Rc value since the increasing Si-NC phase could result in a reduction in the optical gap of the film due to its higher absorption coefficient, as mentioned above (see Figure 4b). To better understand the difference in Jsc among the heterojunction solar cells with various Rc values, losses of the Jsc in the devices were investigated from their IQE data by spectral response measurements. As shown in Figure 5b, the heterojunction device with a higher Rc Si-NCs/SiN x film shows a higher IQE in the short wavelength regime, especially for the wavelength range between 400 and 700 nm, while the IQE spectra in the infrared wavelength regime (>900 nm) are similar for all heterojunction solar cells, implying that recombination of photo-generated charge carriers in the absorber layer is almost the same in all heterojunction devices [26]. Moreover, as depicted in Figure 4a, the obvious variations in the absorption spectra of the P-doped Si-NCs/sc-Si films with various Rc values could be observed at photon energies above 1.8 eV (approximately <700 nm), which shows good correspondence with the trends in the IQE data. Therefore, it is speculated that the difference in Jsc losses among the devices could be attributed to the parasitic absorption in the emitter layer. More photons in the visible spectrum would be absorbed with increasing volume fraction of the Si-NCs in the P-doped Si-NCs/sc-Si film, leading to the limitation in the available solar spectrum in the device, as well as the degradation of the Jsc.
In this report, we have investigated the feasibility of using P-doped Si-NCs/SiN x films as emitters on p-type sc-Si substrates for fabrication of Si-based heterojunction solar cells. From XPS analysis of the P-doped Si-NCs/SiN x films, the P 2p signal only attributed to Si-P or P-P bonds indicates that the P atoms may exist inside Si-NCs. The electrical and optical properties of the P-doped Si-NCs/SiN x material are strongly influenced by its chemical composition (N/Si ratio). The optical gap E04 is enhanced with increasing nitrogen content, while the conductivity is deteriorated. These trends could be interpreted by a bi-phase model, where the SiN x phase contributes to the optical gap enhancement and the Si-NC phase promotes charge carrier transport. Therefore, the Jsc is increased with increasing N/Si ratio in the Si-NCs/SiN x layer, while the FF is reduced. The best cell parameters obtained are Voc of 500 mV, Jsc of 28.2 mA/cm2, FF of 65.2%, and conversion efficiency of 8.6% from the heterojunction solar cell with a Rc = 0.79 Si-NCs/SiN x emitter. Further device optimization is required to improve the photovoltaic efficiency.
This research was supported by the National Science Council of Taiwan under grant nos. 101-2221-E-008-041 and 101-2622-E-008-015-CC3.
Authors' Affiliations
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This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Mistress Lucy's Dungeon Update & Deep Desert Vacation
- 08/22/2013
I know I've made you wait a bit longer than usual for this little update, but I'm sure you all recognize that it's not without good reason…
Since July, a lot has been happening around these parts, and by these parts, I'm referring to My beautiful westside dungeon, Submission LA . That's right pets, Ms. Eden Winter and I have officially decided to join forces behind Submission LA and as such, I will be playing there exclusively for LA-based sessions. This is not to say that I won't be making the occasional house call or taking those of you who need to be shamed publicly on special field trips…it just means that I will no longer offer sessions at My former downtown location.
For those of you who have had the opportunity to experience Submission LA from the inside, I'm sure you can understand My decision to focus my energy there. It is the most elegantly designed, fully-stocked private BDSM dungeon in southern California, and I find its scale and layout to lend itself perfectly for live, intimate training sessions…
In light of this recent development, I invited over a couple Dommes friends and stationed Bob Maverick behind the camera for a test photo shoot. The images turned out beautifully. Here's a little teaser with Ms. Marcella Rose, Serene Sin, and Myself below…you'll find others from that shoot on My newly updated front page.
You're welcome.
Finally, to the subject of this post: I will be on vacation between Aug 25- Sept 4. I will be deep in the desert without phone or email access (gasp!), so I guess you will just have to stare longingly at My virtual image during those eleven dreadful days. If you do need to communicate with Me about something, shoot Me an email and I will be sure to respond upon My return. In the meantime, be good while I'm away painting the town red.


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ASUS announces new ROG Strix X99 Gaming motherboard
ROG Strix is a new series: "The art of expression"
On Tuesday, ASUS announced a new lineup called "ROG Strix," beginning with the ROG Strix X99 Gaming motherboard. With this series in its ROG brand, the company is focusing heavily on dynamic lighting customization and network priority traffic control, two features that will provide gamers and enthusiasts with color-matching RGB lighting effects to inspire PC design creativity, as well as ultra-low latency multiplayer experiences.
ASUS announces new and refreshed X99 Signature series lineup
Fully compatible with Haswell-E and Broadwell-E
On Tuesday, ASUS and its Republic of Gamers (ROG) division jointly announced three new Signature series motherboards built for Socket LGA 2011-v3, featuring ASUS exclusive technologies such as "OC Socket," "Aura" onboard RGB lighting, ThunderboltEX 3, and SafeSlot PCI-Express designs with fortified metal and superior bracket retention mechanisms.
Asus has a new VivoBook out
Intel Braswell Pentium N3700 quad-core inside
ASUS has released a new cheap and cheerful VivoBook powered by an Intel Braswell Pentium N3700 quad-core chip clocked at 1.6 GHz, with Turbo Boost up to 2.4 GHz.
Asus announces new VG278HV 27-inch 144Hz gaming monitor
With 27-inch Full HD TN-panel
Asus has announced its newest 27-inch gaming monitor, which will be based on a Full HD TN-panel with 144Hz refresh rate, the Asus VG278HV.
Asustek reduces its Intel dependence
ZenFone gets Qualcomm and MediaTek platforms
Asustek appears to have pulled back on its Intel dependence for its ZenFones and is adding Qualcomm and MediaTek flavours.
Asus releases VivoStick
At least in Japan
Six months after showing us its Windows computer-on-a-stick, Asus is finally selling its VivoStick – at least to the Japanese.
Asus releases VivoMini PCs
Packed full of Skylake goodness
Asus has announced two VivoMini PCs which show off Intel's sixth-generation Skylake processors.
Asus GTX 980 Ti Strix Gaming Ice spotted
With custom design and Bitspower water block
Asus has rather silently unveiled its new Geforce GTX 980 Ti Strix Gaming Ice graphics card that will be based on a completely custom design and pack a Bitspower full-cover water block for cooling.
ASUS releases GeForce GTX950-2G
Runs under 75W
ASUS has released a new graphics card which does not need a power connector socket.
ASUS quietly launches 28-inch MG28UQ 4K gaming monitor
1ms refresh rate, VESA Adaptive Sync, HDMI 2.0
On Tuesday, ASUS quietly launched the 28-inch MG28UQ, a new 28-inch 4K 3840x2160p 16:9 gaming monitor with rapid 1-millisecond response time to eliminate artifacts and ghosting.


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At a joint press conference with South African President Jacob Zuma this morning, President Obama spoke extensively about Mandela's legacy.
"Our thoughts and those of Americans and people all around the world are with Nelson Mandela and his family and all of South Africans," Obama said. "The struggle here for freedom, Madiba's moral courage, this country's historic transition to a free nation has been a personal inspiration to me, an inspiration to the world, and it continues to be." ...
No, John Y., you're mistaking the President for The Pope and the Papacy.
Ah, Chris, just hold your nose, and look at this particular article, because no I don't give carte blanche blessings to liberal or conservative sites, but anyway, Cost of Obamaâs Trip is Dwarfed By George And Laura Bushâs Africa Spending Spree.


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The Jonas Brothers' new album Lines, Vines and Trying Times will be released on June 16th. In the meantime, fans can check out an exclusive pre-recorded concert — featuring songs from the album and behind-the-scenes interviews — on Walmart.com
To paraphrase Paul Simon, "Every generation throws a boy band up the pop chart." The '60s had The Beatles and The Monkees, the '70s gave us The Jackson 5 and The Osmonds, the '80s begat New Kids on the Block and New Edition, the '90s were dominated by The Backstreet Boys and 'N Sync, while the late '00s are all about the wholesome New Jersey trio of Joe, Kevin and Nick Jonas.
Admittedly, I'm a little long in the tooth to be embracing pop music intended for an audience of teenage girls, but I have to applaud the Jonas Brothers for their musicianship, broad appeal and multi-media dominance. They provide the voices of the cherubs in Night at the Museum: Battle of the Smithsonian, they have their eponymous show on The Disney Channel, and they were up for Best New Artist at the 2009 Grammy Awards. It's a Jonas Brothers world; we're just living in it.
Which of the Jonas Brothers is your favorite? Voice your choice in today's featured poll. And feel free to post a comment with your write-in ballot for younger brother Frankie, the "Bonus Jonas." He's only eight years old, but watch out. Someone's got to own the next decade.
For the latest pop culture news and voting, make sure to sign up for the People's Choice newsletter!


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Arrowsmith, Sue,Quinot, Geo. (Eds.) (2013) Public procurement regulation in Africa /Cambridge, U.K. ; Cambridge University Press,MLA Citation
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Verdi's Requiem to be Performed by Over 200 Musicians at the Stevens Center
8:40am Nov 07, 2013
Giuseppe Verdi turned 200 last month, and the University of North Carolina School of the Arts continues the celebration of his life with a celebration of death: Verdi's monumental Requiem. The Stevens Center performance on Friday night, November 8th at 7:30 PM will feature more than 200 musicians including the UNCSA Symphony Orchestra, Cantata Singers, the Winston-Salem Symphony Chorale, and outstanding soloists from the A.J. Fletcher Opera Institute. One of those dynamic young singers is bass David Weigel. He was joined here by Fletcher Artistic Director Jamie Allbritten.


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Bantwal: Noted author and story-writer Bolwar Mohammed Kunhi has been selected to receive the annual Vishukumar award instituted by Yuva Vahini Kendra Samiti.
Kunhi holds the credit of introducing the lesser-known facets of Muslim way of life to the field of Kannada prose. He has written over 250 short stories in Kannada. Atta itta sutta mutta, DevarugaLa rajyadalli, Anka, Akashakke neeli paradey, Ondu tunDu goDe, Ruqia and RoTTi Patumma are some of his best-remembered works.
Author Upadhya will confer the award on him at a function to be held in the Birwa auditorium at Melkar next week. The award comprises Rs 10,000 in cash, a memento and a citation.


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Testimonial - Cedar Crest Manufacturing Inc.
Dec 31, 2008
"Glenn has been an invaluable partner in our effort to start our company. He has done and excellent job of guiding us through this process and has helped us put together a sound financial plan that we have received excellent reviews from investors. We would not be where we are without the assistance of Glenn."
David Thalacker, Vice President of Operations
Doug Miskimen, President


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Pusha T ft. Ab-Liva - Suicide
It might seem like it was ages ago, but really it's only been a few months since Pusha T was the toast of the town with his solo debut, My Name Is My Name. The calender may have flipped, but Pusha's ability to shake things up certainly hasn't. Now, four months removed from the album releases, Pusha releases a video for the standout cut, Suicide. The edgy, snake-filled, black and white visual matches the cold, ruthless vibe of the track perfectly. Lurking in the shadows, Pusha attacks the dripping Pharrell Williams with his knifing, brash flow. His guest, Re-Up Gang member Ab-Liva, finishes the job with a bare-knuckled, pounding approach. Be sure to grab My Name Is My Name (if you haven't already) and check out Pusha's last Booth appearance on the star-studded TKO (Remix).


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Conan O'Brien to Controversially Officiate Gay Marriage on Show
Conan O'Brien will officiate a gay marriage on his show this week as it moves to New York to celebrate its one-year anniversary on TBS.
Many have accused the event as a ploy for ratings, but the couple is for real. O'Brien will marry a member of his staff and his partner on the program, according a Vulture report.
This week's shows will be taped at New York's Beacon Theater. The show's producers haven't announced what day the ceremony will take place.
This won't be the first marriage to take place on a late night talk show. In 1969, musician Tiny Tim married Miss Vicki Budinger on Johnny Carson's "The Tonight Show." The show broke ratings records while bringing in over 21 million viewers.
The move to New York is part of the celebration for the show's one-year anniversary on TBS. O'Brien's show was broadcast on NBC from 1993 to 2010 after he was paid to leave the network over a scheduling conflict.
The marriage wouldn't have been possible on O'Brien's previous show shot in New York. The state controversially legalized gay marriage when Governor Andrew Cuomo signed the Marriage Equality Act into law this past summer. The change in law brought a wave of protests from Christian conservative groups that believe homosexuality is a sin in the eyes of God.


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Good broke gelding, knows leads well, has a turn on him. Good Ranch Pleasure prospect. Has been trail ridden a lot and hauled to several horse shows and…SOLD
Quiet and good minded gelding. Has 8 months training, very soft in body and mouth, cute body and good bone,lopes and jogs pretty, has a good lead change…SOLD


text: Hi, In the first few lines I see the following... 2007-03-29 15:31:06.045: [ OCRRAW][3076419712]propriogid:1: INVALID FORMAT 2007-03-29 15:31:06.046: [ OCRRAW][3076419712]ibctx:1:ERROR: INVALID FORMAT 2007-03-29 15:31:06.046: [ OCRRAW][3076419712]proprinit:problem reading the bootblock or superbloc 22 2007-03-29 15:31:06.049: [ default][3076419712]a_init:7!: Backend init unsuccessful : [22] 2007-03-29 15:31:06.049: [ CSSCLNT][3076419712]clsssinit: error(22 PROC-22: The OCR backend has an invalid format) in OCR initialization I have no idea what that means, I will see if I can find more information. If you know what that means I would appreciate the help. Crs_stat returns CRS-0184: Cannot communicate with the OCR daemon which makes sense (I think) as CRS is not started. I have not looked at ocrdumpo, I will do that. Bill Wagman Univ. of California at Davis IET Campus Data Center wjwagman@xxxxxxxxxxx (530) 754-6208 -----Original Message----- From: Dyno Fu [mailto:dyno.dba@xxxxxxxxx] Sent: Monday, April 02, 2007 9:26 PM To: William Wagman Cc: Niall Litchfield; oracle-l@xxxxxxxxxxxxx Subject: Re: CRS install failing William, oerr prif 12 000012,0, "failed to initialize cluster support services" // *Cause: The cluster support services module was not configured correctly. // *Action: Check the messages in the log file oifcfg.log. so what is in oifcfg.log? and what is your RAC configuration, i mean, where you put the CSS, OCR file etc. and the output of crs_stat, ocrdump? rgds, Dyno On 4/2/07, William Wagman <wjwagman@xxxxxxxxxxx> wrote: > Niall, > > I've done both of those. Unfortunately I never seem to be able to find > any support people at Oracle who understand or have a knowledge of RAC. > Oh well. > > Thanks. > -- http://www.freelists.org/webpage/oracle-l


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We present the design, synthesis and characterization of new functionalized fluorescent optical switches for rapid, all-visible light-mediated manipulation of fluorescence signals from labelled structures within living cells, and as probes for high-contrast optical lock-in detection (OLID) imaging microscopy. A triazole-substituted BIPS (TzBIPS) is identified from a rational synthetic design strategy that undergoes robust, rapid and reversible, visible light-driven transitions between a colorless spiro- (SP) and a far-red absorbing merocyanine (MC) state within living cells. The excited MC-state of TzBIPS may also decay to the MC-ground state emitting near infra-red fluorescence, which is used as a sensitive and quantitative read-out of the state of the optical switch in living cells. The SP to MC transition for a membrane-targeted TzBIPS probe (C12-TzBIPS) is triggered at 405 nm at an energy level compatible with studies in living cells, while the action spectrum of the reverse transition (MC to SP) has a maximum at 650 nm. The SP to MC transition is complete within the 790 ns pixel dwell time of the confocal microscope, while a single cycle of optical switching between the SP and MC states in a region of interest is complete within 8 ms (125 Hz) within living cells, the fastest rate attained for any optical switch probe in a biological sample. This property can be exploited for real-time correction of background signals in living cells. A reactive form of TzBIPS is linked to secondary antibodies and used, in conjunction with an enhanced scope-based analysis of the modulated MC-fluorescence in immuno-stained cells, for high-contrast immunofluorescence microscopic analysis of the actin cytoskeleton.
Citation: Wu L, Dai Y, Jiang X, Petchprayoon C, Lee JE, Jiang T, et al. (2013) High-Contrast Fluorescence Imaging in Fixed and Living Cells Using Optimized Optical Switches. PLoS ONE8(6): e64738. https://doi.org/10.1371/journal.pone.0064738
Editor: Yingxiao Wang, University of Illinois, Urbana-Champaign, United States of America
Received: February 7, 2013; Accepted: April 18, 2013; Published: June 5, 2013
Copyright: © 2013 Wu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was financially supported by grants from the National Institutes of Health (PN2 EY018241 Nanomedicine Development Center for the optical control of biological function and R01 GM086233-01 to GM and GM086233-01 to YY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Photochemical manipulation of organic molecules has been used for reversible and irreversible control of the two states of photochromic molecules for more than a century [1]–[4], with recent attention shifting towards the design of both synthetic and genetically-encoded photochromes for applications in biology [2], [4]. For example, photochromes that exhibit fluorescence emission in only one of their two states are key to the success of super-resolution fluorescence microscopy [5]–[7], high-contrast fluorescence microscopy (optical lock-in detection, OLID) [8]–[10], and for optical control of protein activity and cellular processes in living systems [11]–[13]. The new optical switches introduced in this study are optimized for high-contrast imaging of ensemble populations of probe molecules in fixed and living cells.
Benzospiropyran-derived photochromes, including 1′,3′-dihydro-1′,3′,3′-trimethyl-spiro[2H-1-benzopyran-2,2′-(2H)-indole] (BIPS) [14]–[17], have been used as optical switches for applications in living cells [8]–[10], [18]–[19]. BIPS undergoes rapid and reversible, high quantum yield transitions between a closed, colorless and hydrophobic spiropyran (SP), and an open, brightly-colored and polar merocyanine (MC), as shown in Figure 1A. Exposure of the MC-state to visible light results in formation of the SP-state, or decay of the MC-excited state to the same MC-ground state, with emission of red fluorescence (Figure 1A) [8]–[10]. The MC-fluorescence while necessarily low because of the competing MC to SP transition, is extremely useful for studies in cells and tissue, as it provides sensitive, quantitative, dynamic and high-spatial resolution read-outs of the two states of the optical switch in the sample [2], [6], currently not possible using azobenzene-derived photochromes [4], [11]. Moreover, since the excited-state transitions between the SP and MC states proceed with defined quantum yields, exposure of a BIPS probe such as NitroBIPS to a defined, alternating sequence of near-UV and visible light results in a time-dependent change in the populations of the two states of the switch, manifest in an intensity waveform of MC-fluorescence [8], as schematized in Figure 1B. On the other hand, the corresponding fluorescence intensity from non-switchable fluorophores, or from background emission in the sample is more or less constant (Figure 1B). The unique properties of optical switch probes have been exploited in the new high-contrast imaging technique of optical lock-in detection (OLID) imaging microscopy [8]–[10]. The modulated fluorescence signal arising from control of a 2-state fluorescent photochrome, or photochromic FRET probe, is isolated from larger "DC"-like background signals in the sample using a digital lock-in detection approach and results in significant increases in signal contrast [8]–[10].
(A), Optically-induced transitions between the SP and MC states of BIPS. (B), Modulation of the MC-fluorescence signal in response to orthogonal control of the SP and MC states.
Unfortunately, most synthetic photochromes are poorly-suited for studies within living cells, as at least one of the two transitions is driven by exposing the cell to <365 nm light [18]–[20], which usually leads to a stress response [21]. While we were the first to show this effect can be minimized by using 2-photon excitation 720 nm to carry out the SP to MC transition in NitroBIPS [9], [12], an easier solution to reduce phototoxicity is to shift the action spectrum for the SP to MC transition to the red >400 nm (or >800 nm for 2-photon excitation) [22]. To this end, we detail a systematic chemical modification strategy in which small π–bonding groups are appended to different sites on the BIPS probe to red-shift the SP- and MC-absorption bands, and further characterized to identify probes that maintain optimal quantum yields for excited state transitions between the SP and MC states, and MC-fluorescence. We also demonstrate the new red-shifted NitroBIPS and their functionalized derivatives are well-suited for rapid optical switching of labeled structures within living cells. These enhancements including an improved analysis for OLID imaging are used as part of a new approach for high-contrast immunofluorescence imaging microscopy.
Rationally-designed Red-shifted BIPS Probes
The strategy to red-shift the SP-absorption spectrum of the BIPS photochrome involves extending π–bond through alkynyl substitution at each locus on the indoline and benzopyran sides of the BIPS molecule [1a–h; Figure 2]. Details of the synthesis and spectroscopic and photochemical characterization of all probes prepared for this study are provided in the (Information S1).
Analysis of the spectroscopic and photochemical data of the new BIPS probes allows us to evaluate the effect of introducing alkyne groups at defined sites on the BIPS molecule on specific spectroscopic and photochemical properties of the SP- and MC-states, the results of which are summarized in Table 1. In general, the absorption spectrum of the SP-state of alkynyl-substituted BIPS is more red-shifted when on the benzopyran side compared to the indoline side, while the extinction coefficient at the maximum SP-absorption wavelength is higher for substitutions made on the indoline side compared to the benzopyran side (Table 1). The SP-state of alkyne substituted BIPS probes (1a–1h) exhibit an intense red coloration upon a brief exposure to 365 nm or 405 nm, followed by a slower thermally-driven (dark reaction) return to the colorless SP-state. The half-life for the thermally-driven MC to SP transition is on the order of 100 seconds in ethanol. This time constant decreases for alkyne substitution made on the indoline side, and increases for alkyne substitutions on the benzopyran side of BIPS. BIPS probes having alkyne groups on the indoline side (1a–1d) have their maximum MC-fluorescence wavelength red-shifted by 5–10 nm compared to 6-NO2-BIPS, while substitutions made on the benzopyran side shift the MC-fluorescence maximum to 650 nm and 680 nm (compounds 1h and 1f respectively). A complete analysis of the spectroscopic and photochromic properties of all alkynyl substituted BIPS identified compounds 1a and 1f (Figure 2), as the most promising candidates for further study. In any case, the alkyne substituted NitroBIPS were converted to their respective anthracene and triazole derivatives in an effort to further extend the absorption maximum of the SP-state without compromising their ability to undergo reversible optical switching and to emit MC-fluorescence.
Triazole and Anthracene Derivatives of BIPS
The alkynyl substituted BIPS probes are readily converted to their corresponding anthracenes, and triazole derivatives. The spectral and photochromic properties of these molecules are summarized in Table 1. Although the SP-states of the anthracene-substituted probes are significantly red shifted compared to alkynyl-BIPS (1a and 1f), only compound 2a exhibits reversible optical switching between the SP and MC states in solution or in living cells. The triazole-substituted BIPS probe, 3f (TzBIPS; Figure 3B) has the most red-shifted SP- and MC-absorption and excitation spectra, and MC-emission spectrum of all the triazole substituted probes (Figure 4). TzBIPS also exhibits robust and efficient orthogonal optical switching between its SP and MC states following alternate exposure to 405 nm (SP to MC) and >550 nm (MC to SP). The quantum yields for transitions between the SP and MC states are notoriously difficult to measure, with the best estimates given as 0.1 [23]. We also note that the quantum yield for MC-fluorescence must be, as we have previously noted [8], necessarily low because of the competing decay of the MC-state from the MC to SP transition. The capacity of the SP-state of BIPS to form the MC-state is quantified by the colorability, which is defined as the absorbance A0 of the solution at the maximum MC-absorption wavelength under a defined irradiation condition (see footnote to table 1). TzBIPS has the highest colorability of all probes prepared for this study with the value at 0.091 (Table 1). Based on these superior properties we selected TzBIPS (3f) and its derivatives for imaging applications within living cells (Information S1).
A), 6-substituted TMS-alkynyl-BIPS (compound 1f); B), TzBIPS, Triazole-substituted BIPS (compound 3f); C), C12-TzBIPS (compound 3fa); D), Synthetic scheme used to prepare triazole substituted BIPS, and the N-hydroxysuccinimide ester derivative (NHS-TzBIPS).
Spectra are recorded for the probe dissolved to 30 µM in ethanol.
High-fidelity Optical Switching of TzBIPS in Living Cells
First, we set about studying the optical switching properties of a cell permeable, and membrane-targeting version of TzBIPS, named C12-TzBIPS (Figure 3C). A single cycle of optical switching between the SP- and MC-states of the C12-TzBIPS in living NBT-II cells (obtained from ATTC) is achieved by first irradiating the yellow-boxed area (Figure 5A) in the field with 5 scans at 405 nm, which effectively triggers the SP to MC transition. The image montage shown in figure 5A shows that the MC-fluorescence is generated from the SP-state only in the region exposed to 405 nm. The entire image field is then exposed to multiple scans at 555 nm, which progressively decreases the MC fluorescence signal in the yellow-boxed region to a very low level (Figure 5C). This reduction in the intensity of the MC-signal is the result of the progressive formation of SP from the 555 nm excited MC molecules, whereas no MC-fluorescence signal change outside of this region. A relatively low power of 555 nm light is sufficient to bring about the MC to SP transition for cells shown in figure 5A (4.5% of the maximum laser output) and is not accompanied by any significant photobleaching of MC, as seen by the constancy of the maximum MC-signal over the 10 cycles (Figure 5C). Subsequent exposure of the yellow-boxed region of the same field to 405 nm results in a repopulation of the MC-state while an additional exposure of the entire field to 555 nm yet again decreases the intensity of the red MC-fluorescence signal. Multiple cycles of optical switching between the SP and MC states of C12-TzBIPS can be brought about in living cells and quantified through measurements of the MC fluorescence signal (Figure 5C). Optical switching between the SP and MC states of C12-TzBIPS in living cells is robust, with little change in the maximum intensity of the MC-fluorescence over the 10 cycles shown in Figure 5C. Moreover, the SP to MC transition is brought about for C12-TzBIPS within living cells using only 10% of the maximum output of the 5 mW laser, and cells are only exposed to 405 nm for 10% of the duty cycle. Modest exposure of the cell to 405 nm light was not accompanied by any noticeable effect on the cell morphology or other indicator evidence of phototoxic effects, for example blebbing, cell-rounding and presence of retraction fibers. In fact the exposure level to 405 nm light is far less than that used to excite CFP fusion proteins, and is comparable to that used to control Lov2-Rac1 in cells [13].
A), Image montage showing MC fluorescence signal of MC-state of C12-TzBIPS within a field of living cells over 3 cycles of optical switching using a low power objective; B), Higher magnification view of the optical switching of MC-fluorescence of C12-TzBIPS within a single cell in the same sample; C), Intensity trace of MC-fluorescence corresponding to the yellow boxed region in (5A); D), Intensity trace of MC-fluorescence within the yellow boxed region in (5B). E), Image montage 6 cycles of rapid optical switching of C12-TzBIPS in living cells within a narrow field of view; F), Intensity trace of the MC-fluorescence averaged over the entire image field (E).
A much faster rate of optical switching of the MC-fluorescence of C12-TzBIPS is realized by reducing the region over which the 405 nm and 555 nm lasers are scanned. NBT-II cells from the same sample used above were selected for a rapid optical switching study (Figure 5 B, D). In particular, a 5 µm×5 µm region within a single cell is exposed to 10 sequential scans of 405 nm at 10% of the maximum laser power. This perturbation generates the MC-state within the selected field and is followed by carrying out 20 sequential scans of the same field at 555 nm (10% of the maximum power), with a pixel dwell time of 1.58 µs. A montage of MC-fluorescence images in the 5 µm × 5 µm region is shown for the first 3 cycles of optical switching (Figure 5B), with a plot of the MC-intensity profile for 10 cycles shown in Figure 5D. The time to complete a single cycle of optical switching is now reduced to ∼1 second again with little evidence of fatigue or photobleaching over the 10 cycles (Figure 5D).
Rapid optical switching of MC-fluorescence in living cells.
Manipulating the SP and MC states of C12-TzBIPS in a smaller region of interest is used to increase the rate of optical switching within labeled cells [13]. Thus by increasing the 555 nm laser power to 20% and reducing the pixel dwell time to 0.79 µs, each image in the field is acquired within 1.89 ms, and a single cycle of optical switching is complete within 7.56 ms. An image montage of MC-fluorescence is shown for 6 cycles of optical switching in Figure 5E, and an intensity trace of MC fluorescence over 10 cycles shown in Figure 5F. As expected, the very narrow dimension of the selected region accentuates the mobility of vesicles, and precludes tracking of individual vesicles between successive frames. Nonetheless, it is evident from the average MC-intensity over the entire field that the MC-fluorescence is modulated within a few milliseconds as a result of rapid transitions between the SP and MC (Figure 5F). Significantly, this demonstration shows that it is now possible to carry out a 4-cycle OLID imaging analysis of a TzBIPS probe within a cell at the video rate.
OLID Immunofluorescence Microscopy
An N-hydroxysuccinimide ester derivative of TzBIPS (Figure 3D) is used to prepare covalent antibody conjugates, which are used as part of an OLID-based approach to improve image contrast in immunofluorescence microscopy. NIH 3T3 cells are fixed using a methanol fixing procedure, and labeled with a polyclonal anti-actin antibody raised in rabbit. After washing off unbound rabbit anti-actin antibody, the cells are treated with the TzBIPS-conjugated goat anti-rabbit antibody. The cells were imaged by confocal microscopy by using a Zeiss LSM 700 microscope [10]. Images of MC-fluorescence are recorded for the same image field throughout the course of optical switching brought about exposing the TzBIPS-labelled preparation to multiple cycles of optical perturbation that includes two scans at 405 nm (15% of 5 mW) immediately followed by 10 scans of the same field at 555 nm (55% of 10 mW).
Enhanced Lock-in Detection Approach to Analyze Modulated Fluorescence Signals
An improved method for optical lock-in detection (OLID) fluorescence microscopy is employed in this study to generate high-contrast images of the modulated MC-emission from TzBIPS labeled actin filaments in cells. In the "scope" approach to OLID imaging [24], we exploit our earlier finding that the 405 nm-mediated change in the fluorescence signal from the MC-state is a sufficiently sensitive indicator of optical switching of the TzBIPS probe in a pixel, and is easily discriminated when analyzed over multiple cycles of optical switching compared to the corresponding response of the signal from background probes and noise. We have introduced both the scope and scope-weighted correlation measures, which are calculated on a pixel-by-pixel basis as described in our previous work [24]. In brief, the scope values are calculated on a pixel-by-pixel basis as follows:
Where, N is the number of cycles used to calculate the scope value, Lmax (x,y)i and Lmin (x,y)i are local maximum and minimum intensity respectively for the nth switching cycle, and (x, y) represents the pixel location. The scope is an average measure of the intensity modulation for the pixel location within a switching cycle. It is computed for each pixel in the image over several cycles of optical switching. For the images detailed in this study, where the noise is very strong and the signal from MC-emission is weak, we introduce a modified scope-based approach, in which a weighted intensity image is constructed using the scope measures as the weights.
OLID analysis of actin in immuno-stained cells.
NIH 3T3 cells are labelled with antibodies against actin and a TzBIPS labeled secondary antibody with a diffuse background fluorescence introduced during chemical fixation. The sample is scanned at 405 nm, which brings about an immediate increase in red MC-fluorescence that is visible upon subsequent excitation of the field to 555 nm. Further scanning of the field with 555 nm reduces the intensity of MC-fluorescence, as seen in the montage of the images of the field recorded over 10 cycles of optical switching (Figure 6A). The MC-fluorescence signal exhibits an identical intensity profile for each cycle of optical switching, and a variance of ∼10% in the maximum signal (Figure 6B). Significantly, the image montage and MC-intensity trace show that TzBIPS experiences little in the way of fatigue or bleaching of MC-fluorescence over the 10 cycles of optical switching (Figure 6A, B). The red fluorescence intensity image of these cells (Figure 6C) recorded immediately following exposure of the cells to 405 nm reveals the presence of actin filaments in stress fibers, cell-surface attachments and at the cell cortex, membrane labeled structures and a diffuse signal throughout the cell that obscures detail of individual actin structures and illuminates other regions indicated with blue arrows that may arise from background introduced during chemical fixation or MC (actin filaments) fluorescence (Figure 6C). The scope measure is computed for every pixel in the image field over the course of the 10 cycles of optical switching, and used to generate the scope-weighted intensity image.
Goat anti-rabbit antibodies labelled with TzBIPS are used to label an rabbit anti-actin antibody previously added to fixed NIH 3T3 cells. A), a montage of MC-fluorescence images over 10 cycles of optical switching within a the immunostained sample harboring a diffuse background signal. The montage suggests that little to no bleaching or fatigue of the TzBIPS probe, which is confirmed in a trace of red fluorescence in a highly modulated region of the image field seen over the ten cycles of optical switching (B). trace of MC-fluorescence signal in a stress fiber over the course of optical switching; (C), image of the red-fluorescence in the sample. Blue arrows reveal loci of background signals that are not visible in the corrleation image. (D), corresponding image of scope-weighted intensity. White arrows indicate regions of the actin cytoskeleton that are well-resolved in the scope-weighted intensity image but not in the red fluorescence intensity image. (E), Pixel intensity profile as a function of pixel position along a line indicated by the orange arrows in figure 6 C,D. The intensities of the actin-rich structure (indicated by the green arrow in figures 6 C,D) and its immediate background are indicated by the green arrows in figure 6 E and used to calculate the signal/background ratio.
Moreover, since the TzBIPS-labelled second antibodies are completely immobilized (by their chemical fixation) over the course of the 10 cycles of optical switching, the value of scope is high and largely uniform within MC-rich regions. Image contrast may be enhanced then by using the scope measures as weights in a scope-weighted intensity image (Figure 6D). This contrast-enhanced image is realized by multiplying the scope measures at each pixel by the corresponding intensity of the fluorescence image (from Figure 6C). The contrast in the scope-weighted image, shown in Figure 6D, is significantly improved compared to the intensity image (Figure 6C), revealing individual bundles of actin filaments at actin stress fibers close to the leading edge of the cell and fine details of cortical actin structures (indicated in white arrows) that are obscured by the background emission in the sample (indicated by blue arrows in Figure 6C). Also apparent in the scope-weighted image (Figure 6D) is the higher contrast in regions away from the cell, the intensity signal of which is evidently dominated by background signals where the corresponding scope value is close to zero, and thus the weighted intensity is almost zero.
Clearly, the scope-weighted image (Figure 6D) leads to a significant improvement in the contrast and noise reduction compared to the original image (Figure 6C). Quantitative measures of the improvement in image contrast between the original and the scope-weighted intensity images (Figure 6C,D) are derived from analysis of the fluorescence intensity profile for a line drawn on the same path for both images (indicated by red arrows in Figures 6C and 6D). The pixel intensity as a function of pixel position is reproduced respectively for both images as shown in Figure 6E. The green arrows in the plot indicate the intensities for an actin rich structure and its adjacent background indicated by green arrows in Figures 6 C,D. The ratio of these intensities provides a measure of the signal to noise in each image as 4.85 and 20.92 respectively, corresponding to an improvement in contrast of a factor of 4.31. Ordinarily, the intensity image of the immune-stained cells reported in Figure 6C would be the one used by an investigator, yet our scope-based OLID approach can be used to improve the signal to background ratio for the image by more than 4-fold. The improvement in image contrast arises largely from the selective amplification of the modulated MC fluorescence signal from the non-switching background fluorescence. In the example shown in Figure 6, the background signal is relatively low and arises from extraneous red emitting molecules created or added to the preparation during the chemical fixation and washing steps. Even greater improvements in image contrast then can be expected in immune-stained samples harboring much higher level of background, such as those derived from thick slices of tissue and intrinsically fluorescent samples such as algae and plant cells. Finally, further improvements in image contrast for samples containing higher background signals than those present in Figure 6 are possible by combining the scope-weighted intensity (used in this study) and the correlation coefficient [8], [24]. Finally, while the focus of our studies using TzBIPS has been on imaging of ensemble populations of labeled proteins and structures in fixed and living cells, it is clear that the optimized optical switching and MC-fluorescence properties of TzBIPS will also find use as probes for ensemble and single molecule imaging beyond the diffraction limit [7], [25]–[27].
We have detailed the rational design, synthesis and characterization of new fluorescent optical switches based on the BIPS photochrome for all-visible light driven control of MC-fluorescence in living systems. These spectroscopic and photochemical analyses are used to identify a new class of BIPS photochrome, called TzBIPS whose SP and MC absorption spectra, and MC-fluorescence are the most red-shifted of all known BIPS derived photochromes [3], [18]–[20]. A membrane-directed C12-TzBIPS probe is shown to undergo rapid and reversible, high-fidelity optical switching between the SP and MC states in response to alternate exposure of the cell to low powers of 405 nm and 555 nm lasers. Optical perturbations carried out over 10 or more cycles of optical switching do not cause any noticeable fatigue or bleaching or stress-like effects on the cell. The progress of optical switching is quantified in these living cells by recording images of the 555 nm induced MC-fluorescence. The 405 nm triggered SP to MC transition of C12-TzBIPS is complete within the 790 ns pixel dwell time of the confocal microscope, which suggests that the photochemistry reaction underlying the SP to MC transition is most certainly faster, and is not a time-limiting factor for optical switching. A single cycle of optical switching of C12-TzBIPS in a region of interest is realized within 8 ms (125 Hz) within living cells, the fastest of any reported BIPS probe. This property allows the background fluorescence in a sample to be corrected (subtracted by subtracting the post from the pre-405 nm exposed MC-fluorescence images) much faster than the video-rate, a unique capability for probes used within living cells. We note that the rate of optical switching of MC-fluorescence could be improved even further by decreasing the time required to switch between scanning the field with the 405 nm and 555 nm lasers. Finally we introduce an amino-reactive form of TzBIPS for covalent labeling of biomolecules, including secondary antibodies that are used in this study as part of an approach for high-contrast IF imaging microscopy. The scope-weighted intensity analysis is particularly well-suited for high-contrast imaging of chemically-fixed TzBIPS probes, and can realize improvements in the signal to background ratio of >4-fold improvement, even for samples that are otherwise considered background free. The new TzBIPS labeled antibody simplifies the design of antibody probes for OLID-IF compared to FRET-based antibody probes [10], [28], and will be useful for IF preparations containing high levels of intrinsic fluorescence, and for the imaging of labeled cells within thick tissue.
Supporting Information
Information S1.
Details of the synthesis, spectroscopic and photochemical characterization of all the probes.
Author Contributions
Conceived and designed the experiments: LW GM YY. Performed the experiments: LW XJ YD CP JEL YY GM. Analyzed the data: LW YD CP XJ JEL TJ YY GM. Contributed reagents/materials/analysis tools: LW YD XJ GM. Wrote the paper: GM YY LW.
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With only three episodes left, Hostages is slowly preparing for its endgame: the assassination of the president.
Meanwhile, Brian goes to the police to expose Duncan. But when Burton shows up showing a photo of Ellen and Duncan kissing, Brian decides not to tell the truth. Instead, he asks Morgan to steal a phone from a friend, and later calls Nina to tell her the truth. Nina is shocked to learn the truth, and in the ends decides to confront her husband.
Elsehwere, Ellen is getting back to normal when she has surgery. It is nice to see some focus on the Sanders again. Today, Ellen is operating on a criminal who killed a cop. While one of the nurses is ready to give up on the murderer, Ellen pulls through and eventually saves his life. After this, she decides that she cannot kill the president. This will undoubtedly have far-reaching consequences for her and possibly her family.
As we are getting closer to the end of the series, things are getting closer together and plans are falling apart. We are all curious to see where it leads and what will eventually happen. This episodes sets things nicely in motion for the last few episodes.
What did you think of this week's episode? Promo for next week:


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Opal Irene (Ebie) Boone
April 20, 1923 - July 9, 2013
Twain Harte resident Opal Irene (Ebie) Boone died July 9 at Memorial Medical Center in Modesto. She was 90.
Mrs. Boone was born in Empire to John and Molly (Smith) Ebie.
She married Virgil Leroy Boone in 1941 and they lived in Empire for a number of years before settling in Twain Harte 56 years ago. They moved to the area because it suited their son, John L. Boone's, allergies.
She worked as a secretary and auditor for the Tuolumne County Water District No. 1 (now the Twain Harte Community Services District) for 19 years until her retirement in 1988.
She served on the Twain Harte Park and Recreation Board for a number of years and was instrumental in developing Eproson Park.
She enjoyed reading, and doing crossword and jigsaw puzzles.
Mrs. Boone was preceded in death by her husband, Virgil Leroy Boone, in 1990.
She is survived by her children and their spouses, Carole and Tony Reyes, of Twain Harte, John Leroy Boone, of Twain Harte, and Pat and Daniel Hillier, of Tuolumne; her grandchildren and their spouses, Joe and Gwen Reyes, of Oakhurst, William and Tracy Hillier, of Salida, Rebecca Anderson, of San Rafael, Jonni Molgaard, of Redding, Nikki and Dan White, of Cottonwood, Jeremy and Elizabeth Boone, of Twain Harte; sister, Ruby Bishop, of Modesto; and numerous great-grandchildren and great-great-grandchildren.
No services are planned.
Neptune Society of Central California is handling arrangements.
Naomi (Rawlins) Erickson
June 15, 1917 – July 10, 2013
Former Tuolumne County resident Naomi Erickson died July 10 at Rideout Memorial Hospital in Marysville. She was 96.
Mrs. Erickson was born Naomi Rawlins in Independence, Ore., to Benjamin and Catherine Rawlins.
Her family traveled by covered wagon to Escalon in 1920. They later settled in Modesto. She attended school in Modesto and graduated from high school in Salida.
She worked as an electrologist in McHenry Village in the early 1950s.
She married John Beiler, of Modesto, in 1934. They later divorced.
She became a rancher's wife when she married Paul Erickson, of La Grange, in 1955. She learned to ride horses as well as cook for cattle drives and brandings.
After her husband's death in 1972, she briefly lived in Guadalajara, Mexico, and also in Oklahoma and Texas, before returning to California.
Mrs. Erickson moved to Twain Harte in 1955 and later settled in Soulsbyville.
She moved to Lake Havasu, Ariz., in 2011 to live with her niece and to Yuba City in 2012 to live with her son.
Mrs. Erickson was a member of the Calaveras-Tuolumne Cowbelles and St. James Anglican Church.
She enjoyed the ocean, traveling, music, and art and was a museum docent in Stockton during the 1970s.
Mrs. Erickson was preceded in death by her husband, Paul Erickson, in 1972; daughter-in-law, Joanne Beiler, in 2009; siblings, George Rawlins, in 1978; and, Ruth Hoover, in the mid-1990s; grandson, Gary Crauthers, in 1969.
She is survived by her children and their spouses, Bert Beiler, of Yuba City, Tim and Bette Ann Erickson of, La Junta, Colo., Paula Crauthers, of Soulsbyville, and Betty Varain, of Turlock; 11 grandchildren; 11 great-grandchildren; 14 great-great-grandchildren; and one niece.
A graveside service will be held at 1 p.m. Saturday at Lakewood Memorial Park, 900 Santa Fe Ave., Hughson.
Heuton Memorial Chapel is handling arrangements.
William Herold Taylor 'Billy' Suderman
Aug. 7, 1965 - April 27, 2013
Former Tuolumne County resident William Herold Taylor "Billy" Suderman died April 27 in Stapleton, N.Y. He was 47.
Mr. Suderman was born in Emmett, Idaho, to Diane and William H. Taylor.
He was raised in Tuolumne County and graduated from Sonora High School in 1983. He earned a bachelor's degree in art history from the University of California, Santa Cruz, in 1987, a master's degree from Bordeaux University in France, and a doctorate from Pantheon-Sorbonne in Paris, France.
He taught art history at universities in Paris and San Juan, Puerto Rico, and the Snug Harbor Cultural Center and Botanical Gardens in New York City, N.Y.
Mr. Suderman was a member of the Unitarian Church of Stapleton. He was also a member and advocate of the New York City lesbian, gay, bisexual and transgender community and volunteered with a New York hospice program.
Mr. Suderman was preceded in death by his parents, Diane and William H. Taylor.
He is survived by his partner, Michael Fielding, of Stapleton; brother, Sean Taylor, of Nevada; and stepmother, Cathie Peacock, of Long Barn.
In lieu of flowers, donations can be made to Hospice of the Sierra, 20100 Cedar Road North, Sonora CA, 95370.
A memorial service will be held from 11 a.m. to 2 p.m. Saturday at 21761 Cedar Springs Road, Twain Harte.
Russell Robert Willard
Jan. 14, 1924 - July 4, 2013
San Andreas resident Russell Robert Willard died July 4 at his home. He was 89.
Mr. Willard was born in Iowa to Robert and Flossie Willard.
He moved to California as a young man and served in the U.S. Marine Corps during World War II.
After his service, he worked at Western Pacific Railroad and later as a rancher in Stoneyford.
Mr. Willard worked as a plumbing contractor from 1965 until his retirement in 2012.
He lived for a number of years in Carmichael, Sacramento County, and Stanley, Idaho, before settling in San Andreas 30 years ago.
He was an athlete and sportsman and enjoyed building things.
Mr. Willard was preceded in death by his wife, Laurel Willard, in 1968; and grandson, Shayne Willard, in 2007.
He is survived by his partner of many years, Pat Seidler, of San Andreas; his children, David Willard, of San Andreas; and Denis Willard, and his wife, Carol, of Orangevale; granddaughter, Monique Stewart and her husband, Clint, of Sacramento; and two great-grandchildren.
A memorial service will be held at 11 a.m. Saturday at San Andreas Memorial Chapel, 254 W. St. Charles St., San Andreas. A reception will follow at Joe Cardenas' house. Maps will be provided.
San Andreas Memorial Chapel is handling arrangements.
BURFORD - Fay Earleen Burford, 75, of Arnold, died July 5 at her home. A memorial service will be held at 6:30 p.m. Thursday at the Danville Congregational Church, 989 San Ramon Valley Blvd., Danville. A memorial service will be held on Aug. 10, in Arnold, with time and location to be announced. Angels Memorial Chapel is handling arrangements.
RASMUSSEN - James Rasmussen, 60, of Soulsbyville, died Monday at Sonora Regional Medical Center. Terzich and Wilson Funeral Home is handling arrangements.
STEFFEN - Frances Steffen, 97, of Sonora, died Tuesday at Avalon Care Center of Sonora. Terzich and Wilson Funeral Home is handling arrangements.