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Topaz received the "Deal of the Year" Award at the 2015 Seatrade Global Awards in London, for its contract win with BP Exploration ("Caspian Sea") Limited ("BP"), which was announced in March 2016.
Read Topaz and BP sign long-term contracts for 14 ships in Azerbaijan
René Kofod-Olsen, Chief Executive Officer, Topaz Energy and Marine, said of the award, "We are honored to have been awarded the Seatrade 'Deal of the Year' Award. This award marks the first time Topaz has secured a prestigious international award and reflects our increasing presence as a globally significant OSV business. We are especially proud to have been recognized for our efforts in a period of tough market conditions. We are grateful to have the opportunity to work with world-class clients such as BP and we thank them for their ongoing trust in our ability to deliver."
The Seatrade Awards were first introduced in 1988, to celebrate and recognize the achievements of the industry and outstanding individuals within it. This year sees the 28th edition of the awards.
COO Geoff Taylor (second from left) received the Deal of the Year Award on behalf of Topaz
Topaz has scooped two awards at the 2015 Lloyd's List Middle East and Indian Subcontinent awards in the categories of 'Offshore and Energy' and 'Maritime Services'. Topaz was also Highly Commended for the Safety Award.
The awards recognise Topaz's excellence in providing services to the offshore industry with entries judged on elements such as customer service, safety, environmental protection, cost management as well as vessel efficiency and reliability and company profitability.
Topaz's long-term strategic focus of maintaining a younger fleet drives safe, environmentally-friendly and cost-effective services, resulting in higher utilization, which currently sits at 87% despite challenging market conditions. Its excellent safety track record, capital discipline and excellent operational health and safety track record have helped entrench its MENA and Caspian stronghold and facilitate its expansion into new markets in Africa.
René Kofod-Olsen, CEO, Topaz Energy and Marine, said:
"It is an honour to be selected for not one but two awards at the highly regarded Lloyd's List awards event among a highly competitive peer group.
"At Topaz, we pride ourselves on our young and technologically advanced fleet, our model of securing medium to long-term contracts and our ability to actively manage costs in a difficult operating environment. These awards provide external recognition of our efforts in providing best-in-class services to the offshore industry and I would like to dedicate the awards to our seafarers working across the MENA, Caspian and African regions who dedicate themselves every day to excelling in our industry."
From left to right: CEO René Kofod-Olsen, Head of QHSSE Ian Trebinski, and COO Geoff Taylor
Topaz has been selected as the winner for 'The Safety and Quality Award' at the 2015 Seatrade Maritime Awards amidst very strong competition. The award recognised the Company's uncompromising focus on safety and reputation among clients for maintaining a consistently strong safety performance over its 40-year history.
René Kofod-Olsen, CEO, Topaz Energy and Marine, said:
"We would like to thank the judges and organizers of the Seatrade Maritime Awards for selecting us as winner for 'The Safety and Quality Award.' As the premier maritime awards for the Middle East, Indian Subcontinent and Africa, it is an honour to be selected amongst such a strong peer group.
Safety is our absolute top priority and we are therefore delighted that we have been recognised for the work we have done to achieve high safety standards. We will continue to invest in innovative solutions to help us improve safety and quality which in turn helps drive our long-term sustainable business plan in the region.
CEO René Kofod-Olsen accepted 'The Safety and Quality Award' on behalf of Topaz.
Topaz has won the Offshore and Energy Award and The Ship Operator Award and was Highly Commended in The Safety Award category at the Lloyd's List Middle East and Indian Subcontinent Awards 2014.
The Offshore and Energy Award recognised Topaz's excellence in providing services to the offshore energy industry and The Ship Operator Award recognised Topaz's excellence in the operation of its ships over the past year. In addition, Topaz was highly commended in the The Safety Award.
René Kofod-Olsen, CEO, Topaz Energy and Marine, said: "We pride ourselves on offering cost-effective quality services to the highest standards of safety. We are therefore delighted to have our efforts recognised by Lloyd's List. Winning these prestigious awards would not have been possible without the dedicated effort by all our employees and they deserve the highest praise and appreciation."
CEO René Kofod-Olsen accepting the 'Offshore and Energy' award on behalf of Topaz
Topaz has been selected as the winner for 'Offshore Marine Development – Africa' at the 2014 Seatrade Maritime Awards. This award recognized Topaz's achievement in overcoming challenges and its significant investments and contributions to the African offshore market in the last year.
René Kofod-Olsen, CEO, Topaz Energy and Marine, said: "Our team could not be more deserving of this recognition considering the fast growth of our West African business in line with our Africa strategy set in 2012. The region presents significant challenges and we're very pleased with our progress in partnership with our clients and stakeholders."
CEO René Kofod-Olsen accepting the Seatrade 'Offshore Marine Development - Africa' award.
Topaz Energy and Marine won the Lloyd's List 'Offshore and Energy' award at the recently held Lloyd's List Middle East and Indian Subcontinent Awards at the Grand Hyatt in Dubai. The prize was awarded for excellence in providing services to the offshore energy industry via the operation of offshore support vessels.
Accepting the award on behalf of Topaz was Chief Executive Officer, René Kofod-Olsen, who commented, "We are honoured to receive this award as it recognizes the reliability of our services, our strong commitment to safety and the sustained growth of our fleet with focus on modern and technologically advanced tonnage".
Topaz was also a finalist for the 'Ship Operator' award.
CEO René Kofod-Olsen accepting the 'Offshore and Energy' award on behalf of Topaz
Topaz won the 'Workboats' award at the recent Seatrade Middle East and Indian Subcontinent Awards held at Atlantis, The Palm in Dubai.
Accepting the award on behalf of Topaz was Chief Operating Officer, Roy Donaldson, who commented, "It brings us great pride to be recognized in the industry through this award for our commitment to operational excellence and safety and our focus on maintaining a young and technologically advanced fleet".
Roy Donaldson (center) accepting the 'Workboats' award on behalf of Topaz. Photo courtesy of Seatrade.
Topaz Energy and Marine was awarded the 'Ship Operator Award' at the Lloyd's List Middle East and Indian Subcontinent Awards 2012.
Commenting on the occasion, Roy Donaldson, Chief Operating Officer, Topaz Energy and Marine said, "Topaz Marine are proud to be the 2012 recipients of the prestigious Lloyd's List Ship Operator of the Year Award, which further cements our commitment to the highest standards in operating quality."
Topaz was also highly commended in the 'Safety Award' category.
COO Roy Donaldson receives the Lloyd's List 'Ship Operator of the Year Award' on behalf of Topaz
Topaz Marine was awarded the prestigious 'Workboats Award' for the fifth successive year amidst strong competition at the Seatrade Middle East and Indian Subcontinent Awards 2012 held in Dubai. The award is based on a number of criteria including, commitment to safety, technological advances, strength of client base, commitment to customer service, current & future growth initiatives, innovation, commitment to training of personnel and corporate social responsibility. Being the recipients of this prestigious award year on year is proof that Topaz has consistently raised its bar of performance and maintained its leading position in the Oil and Gas Marine services. Further to Topaz's commendation at the Lloyd's List Middle East and Indian Subcontinent Awards 2012 in October for the "Safety Award", Topaz Marine was also shortlisted for the Seatrade "Safety and Quality Award" this year.
COO, Roy Donaldson (centre), receives the 'Workboats Award' in behalf of Topaz Marine at the Seatrade Middle East and Indian Subcontinent Awards 2012
Topaz Energy and Marine won two awards out of four nominations at the annual Seatrade Middle East & Indian Subcontinent Awards 2011 held in Dubai.
Stephen R. Thomas, commented on the awards, saying, "The Seatrade Awards are a benchmark for excellence in our industry. We are very honoured to have won these two awards amongst other first-class finalists. We shall treat the Safety & Quality Award as a charge of responsibility to maintain the highest standards of safety in all that we do."
Receiving the awards on behalf of the division was Roy Donaldson, COO of Topaz Marine, who said, "Our people work tirelessly to maintain a spotless safety record, and winning the Safety & Quality and Workboats Awards demonstrates the unwavering commitment and serious attention we place on safety and efficiency across our businesses. This has brought great honour to the company and we could not be more pleased by the outstanding effor of all those in Topaz Marine to qualify in four major categories.
Topaz's awards were won in strong competition with prominent companies and executives of the maritime community in the region and the Indian subcontinent ar a glamorous ceremony organised by Seatrade organisation. Topaz was also represented as a finalist in the 'Technical Innovation' and 'Marine Offshore Services' categories.
Safety & Quality Award - Topaz's commitment to safety permeates all levels of the organization, and the company always strives for an incident-free workplace through an uncompromising approach to safety. A systematic training and empowerment program ensures that all stagg actively participates in the development of ongoing improvements to an already well-established safety system.
Workboats Award - Operating a diverse fleet of vessels, Topaz Marine offers a wide range of offshore support services, including towage, rig moves, pipelay support, survey, subsea development, installation maintenance and repair, production and development supply, cable laying, ice breakers, etc. With our modern and dynamically positioned fleet, Topaz Marine is well-placed to offer quality, safe and reliable service to its customers.
Topaz Marine COO, Roy Donaldson, receiving 'The Safety & Quality Award'
Topaz Marine COO, Roy Donaldson, receiving 'The Workboats Award'
Following the first win in 2010, Topaz Marine MENA has won the coveted RoSPA silver award for 2011. RoSPA, The Royal Society for the Prevention of Accidents, is a registered charity with a core focus on accident prevention in the UK and the rest of the world.
The RoSPA Silver Award
Topaz Energy and Marine was presented with two prestigious honours at Lloyds List Middle East & Indian Subcontinent Awards 2010. The awards ceremony, held on Wednesday 10th November in Dubai, saw Topaz Marine MENA accept the 'Ship Owner and Operator' award and the 'Energy' award.
Topaz Marine MENA was recognised by the judges for enhancing customer service even during the downturn leading to an increased customer base and also for Topaz's restructuring efforts, which modernised the fleet and allowed for the provision of a more prompt and comprehensive service to the offshore industry.
Ron Clark, General Manager of Topaz Marine MENA, commented: "To have won three awards in one year was already an outstanding achievement so now to add a further two is absolutely amazing. My thanks go to all of the Topaz Marine MENA staff and our crews. These awards truly belong to the entire Topaz Energy and Marine group and we will be very proud to display them on behalf of the whole Topaz family".
Topaz wins the 'Ship Owner and Operator' and the 'Energy' Awards
Topaz Energy and Marine were proudly awarded three significant accolades at the 2010 Seatrade Awards including 'Shipping Company of the Year', 'Marine & Offshore Services' and the 'Workboats' award.
Topaz was also a finalist in the Ship Repair/Shipyard category and the 'Corporate Social Responsibility' category for its work with the 'Flying Angel', a seafarer's support boat operating in the offshore anchorage of Fujairah.
Topaz Marine COO, Roy Donaldson, receiving 'The Marine and Offshore Services' Award
Topaz Marine MENA won the Lloyd's List Middle East and Indian Subcontinent 'Safety at Sea' award. The triumph was motivated by Topaz maintaining the highest standards in safety in their daily operations and making concerted efforts in training and investment in high-tech safety solutions. Topaz's Lloyd's List awards were won in competition with prominent companies and individuals of the maritime community in the Middle East and India. Topaz Marine MENA has won several Lloyd's List awards in recent years and was a finalist in three other categories of the evening; 'Energy', 'Innovation' and 'Ship of the Year'. Accepting the award on behalf of Topaz Marine MENA was GM, Ron Clark, "This honour belongs to our entire staff and especially our HSE team for the great work they do in pushing the envelope on safety. The safety culture permeates Topaz's whole organization and to be recognized for that is truly encouraging and inspires further progress in this pivotal area", said Clark
Topaz wins the 'Safety at Sea' Award
Topaz Marine MENA won a Seatrade award for the third consecutive year. The 'Workboats' award was secured thanks to Topaz's fleet size, diversity and low average fleet age. GM of Topaz Marine MENA, Ron Clark commented on the award, "It's a great honor to have won at Seatrade for the third year running, that's something I'm very proud of. This win would of course not have been possible without the commitment of our excellent staff and management of whom I'm truly grateful".
Topaz wins the 'Workboats' Award
Topaz subsidiary, Nico International won the Ports & Maritime category of the Dubai Environment Health and Safety Award on behalf of Trakhees Environment, Health & Safety, Ports, Customs & Free Zone Corporation. The Awards were held under the patronage of His Highness Sheikh Hamdan Bin Mohammed Bin Rashid Al Maktoum. It is the second consecutive year that Nico International has won this award.
John McFadyen, GM Topaz Marine Repair and LG Iyer, QHSE Manager, received the EHS Award
Topaz subsidiary, Nico Middle East won Seatrade's Workboats Award for the Middle East and Indian Subcontinent.
Topaz Marine MENA wins the 'Workboats' Award
Topaz subsidiary, Nico Middle East won Lloyd's List Innovation and Safety Award for the Middle East and Indian Subcontinent.
Topaz wins the 'Innovation and Safety' Award
All three business units of Topaz Marine are ISM (International Safety Management) and ISO 9001 certified.
BP 'President's Award' for safe recovery of CA platform personnel
Copyright 2017 Topaz Energy and Marine.


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We are
through this matrix of 12 archetypes, i.e. Zodiac constellations.
Well of the last 2000 years unknowingly to many, there are 13 archetypes.
Plus 2000 yrs ago, man fixed the zodiac like a calendar, the problem with that is the zodiac changes with time. 80% of astrology sites give the wrong birth signs. Astronomy is the key today to knowing a true birth sign, there are Ophiuchus' out there and they dont even know they are such.
Interesting Ophiuchus is symbolized by a man holding a snake, There is much said about a sepent race, perhaps this is where they come from. I have no clue, I just looked up the word Ophiuchus and found this out.
Do you know anything about this?
Ive read many things, is there truth to any of it IDK.
I know that the sun rises in Ophiuchus towards the very end of Nov. until about Dec. 17th/18th...I could get better dates if you want specific. This also means that all other signs have changed in that their time of the sun in the sky each year varies over time.
But it is clear to astronomy, that if we are viewing the sun and sky from Earth, that the sun signs change over time due to our wobble.
If it is true that signs are what we project through...I think we need to know where the sun is really at when one is born...and not where the sun would of been a couple thousand years ago.
There are all sorts of videos about Ophiuchus. I look at it as the time of choice or ones who will hold a vibration leaning towards defiantly positive or defiantly negative. They hold the vibration for others to make a choice...do we handle the snake and tame chaos...or do we release the snake, releasing chaos.
But archetypes are personal to every one...so what it truly feels to another can be something totally different to someone else.
Some will say its the coming of Satan, some will say its the healer.
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Israel, a startup oasis in the middle of a war zone
Israel — the Startup Nation — has perfected the art of not just surviving but flourishing in it. Today, this nation of beaches is considered a startup haven.
Currency futures investors find oasis in Dubai
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Oasis Centre Gets Rs 40 crore from Stake Sale
Oasis will also invest the proceeds to procure most advanced reproductive technologies and equipment to offer unique infertility treatments.
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One of the attackers cursed a diner for sitting with non-Muslims during Ramadan and proclaimed that the nation would now be seen as an Islamic state.
Amazon Kindle Oasis review: Does not offer more than Paperwhite
Kindle Oasis is the company's slimmest and lightest Kindle with an all-new square design instead of the usual rectangle.
A creative oasis: The Museum of Goa is so much more than just a gallery
MOG is simple yet elegant three-storey white structure for photographs, paintings and installations, besides other art forms like music, cinema, dance.
Oasis Group to invest Rs 500 crore for setting up ethanol plant
Oasis Group is planning to invest Rs 500 crore for establishing a wheat-based ethanol plant with capacity of 5 lakh litres per day at Ferozepur in Punjab.
India oasis of growth in barren eco terrain: DIPP Secretary Amitabh Kant
India will see many young entrepreneurs become part of this supply chain and there will be several startups in defence manufacturing, which will bolster Make-in-India.
Vasant Oasis buyers file complaint against developer
The buyers have also made a nominee director of MSREI, financial partner in the project, a party to their petition, which has now been admitted.
CIIE joins hands with Startup Oasis, RIICO to launch accelerator programme for startups in water sector
The programme is set up in partnership with Hindustan Unilever's Pureit, Arghyam Foundation, Luminous Water Technologies' Livpure, the World Bank's Water and Sanitation Program, and the Asian Development Bank.
Dubai Food Festival 2015: Culinary oasis in a desert
Some of the culinary treats can be experienced round the year. If you travel to the city, these are the experiences you should definitely try out.
India an oasis of tranquility amid Greek tragedy, China carnage? Here's what to bet on
Dalal Street is of the view that there are no re-rating triggers domestically, and the consolidation phase could extend further.
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Euro zone needs growth and austerity: ECB's Asmussen
By Annika Breidthardt
BERLIN (Reuters) - A European growth pact and deeper political integration in the euro zone could bolster the currency union, but there must be no softening of the bloc's fiscal pact on budget discipline, ECB policymaker Joerg Asmussen said on Monday.
Presenting a vision for Europe over the next 10 years, Asmussen, who joined the ECB's Executive Board this year after serving as German deputy finance minister, said the right crisis response was "not less, but more Europe" and that fiscal union was ultimately the right path.
There is a growing push in the euro zone, led by newly elected French President Francois Hollande, to do more to stimulate growth and not just focus on reducing deficits - German Chancellor Angela Merkel's mantra.
Asmussen said the debate on growth versus austerity was the "wrong debate" to hold.
"We need both," he said in a speech in Berlin, though he stressed the need for fiscal consolidation and reforms.
"The fiscal compact can be complemented by growth-enhancing measures. This makes sense as a supplement, but the fiscal compact cannot be renegotiated or softened," he said.
The ECB has helped fight the debt crisis by cutting interest rates to a record low of 1 percent, flooding financial markets with more than 1 trillion euros in three-year loans and buying struggling euro zone countries' bonds in the secondary market.
Asmussen stressed that the ECB's non-standard measures were of a temporary nature and could be withdrawn any time if inflation risks emerged. Continued...


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Mathura: Tibetan spiritual leader the Dalai Lama would be visiting the city on Monday and will be staying at the Karshni Ashram Raman Reti here, an official said today.
Almost 1000 representatives from 35 countries around the globe are attending the Conclave.
Buddhist monks and scholars from various countries are participating in the seminar "Buddhism in 21st Century" being held at International Convention Centre here, about 100 km from the capital Patna.
India deliberately used the Dalai Lama, who is not only a religious figure but also a political one, as a "legal excuse" to claim the disputed region, said Hu Shisheng, the director of the Institute of South and Southeast Asian and Oceania Studies.
He emphasised the need for refinement of emotions while adding that compassion was as much the make of human beings as violence.
Previously, some global Buddhist conferences barred the Dalai Lama because of China's opposition.
Rick Ross: "I Told Meek I Wouldn't Trust Nicki Minaj"
Since the two split earlier this year, Meek believes he's been the subject of scathing headlines about his personal matters. A week ago, Meek referenced the lyrics in a post on Instagram with the caption: "Rozay been told me not to trust you".
Tibetan spiritual leader Dalai Lama and other dignitaries released a Pali Tripitaka book at the inaugural session of the Conclave. Inaugurating three day worldwide conference on secularism here, he said India was a confluence various religions and cultures and it was the only country in the world, which was spreading the message of world peace and global brotherhood by strictly adhering to the principles of secularism. "China is strongly dissatisfied and resolutely opposed to this", Ms. Hua observed.
"There is a lot of expectations from Buddhism across the world", the Minister said.
The Dalai Lama lit a traditional lamp to mark the start of the meeting at Rajgir, a Tourism Department official said.
China was also peeved when President Pranab Mukherjee hosted the Dalai Lama at his official residence with other Nobel prize winners in December past year.
Terming the yatra as praise worthy initiative of the State Government, Dalai lama has supported the Narmada Seva Yatra being set out to conserve Narmada river.
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The hotel was opened in 2005 and comprises a total of 179 rooms including 2 suites and 2 rooms with disabled access spread over 4 floors. Facilities include a foyer with a 24-hour reception desk, a safe, a currency exchange bureau, a hairdresser's salon, a small supermarket and a selection of shops. Dining options include a bar and a restaurant with highchairs for infants and younger guests may let off steam in the playground as well as participate in kids' club activities. Laundry, room and medical services are available and those arriving by car may make use of the car park and garage facilities.
This hotel lies in the Camyuva district of the city just 6 km from the centre of Kemer. A diverse range of shops and entertainment venues are to be found in the near vicinity, as are restaurants, bars and exciting nightlife venues. It is 100 m to the nearest beach and transfer to the airport takes about 1 hour.
Each of the hotel's pleasant rooms comes with an en suite bathroom with a hairdryer, a balcony or terrace, a direct dial telephone, satellite/ cable TV, a minibar, a king-sized bed, carpeting, a hire safe and centrally controlled air conditioning.
In the outdoor complex is a swimming pool and children's pools, a poolside snack bar and a sun terrace with sun loungers and parasols laid out ready for use. Further leisure options include a sauna area, steam baths and various spa and massage treatments. Sports enthusiasts may play table tennis, beach volleyball, billiards, and aerobics as well as make use of the in-house gym.
Meals are served in buffet form.
All meals are available in a buffet form; special dietary requirements and individual-specific dishes can be catered for. It is possible to book an all-inclusive stay.MasterCard and VISA are accepted as methods of payment.
Some facilities may not be operational due to weather / seasonal conditions.
Room Facilities:
Direct dial telephone
Satellite cable TV
Internet access
Double bed
Central airconditioned


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"For the past ten years, due to stress at work and home, I have lost my eyebrows. I have been pencilling my eyebrows every morning for the past 10 years and that added to my daily stress. I have never been able to swim under water, or afraid of being splashed in the face; also'Jhe constant worry of perspiring at the gym and not being able to wipe my brows! you can just imagine what I have been going through!
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– Brenda P.
Testimonials appearing on this site are actually received via e-mail, or written submission. They are individual experiences, reflecting real life experiences of those who have used our products and/or services in some way or another. However, they are individual results and results do vary. We do not claim that they are typical results that consumers will generally achieve. The testimonials are not necessarily representative of all of those who will use our products and/or services.


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Posted on Jun 7
Wood Dale, IL
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This ENTRY LEVEL position will work with our Engineering team and create new drawings, update existing drawings, review detail, check and assist in release of development and production drawings. Validate part creation from concept to production print. Validate and document assemblies and piece parts per company and industry standards for drafting, material specifications, manufacturing processes, etc.
Direct hire, full-time opening with benefits package including overtime (when required), health insurance, paid vacation, profit sharing, 401K.
Advance Thermal is a premier manufacturer of removable insulation covers in North America. With three fully equipped manufacturing facilities located across North America, quality insulation blankets are manufactured for a wide array of industrial manufacturing equipment and utilities. Advance Thermal Corp. is currently seeking a Mechanical Designer in our Wood Dale, Illinois headquarters.
Evaluates mechanical systems and products by design and research.
Confirms system and product capabilities by design feasibility and testing.
Develops mechanical products by studying customer requirements and soliciting observations from field service.
Assures system and product quality by designing testing methods
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College coursework in design software; AutoCAD, SolidWorks,and Microsoft Office including Excel or equivalent work experience using the software.
Able to follow job specifications & standards/able to recognize a flawed design. Ability to read machine drawings and customer part drawings
Strong analytic and problem solving skills.
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Definition of 'empty-headed'
If you describe someone as empty-headed, you mean that they are not very intelligent and often do silly things.
...a pretty, empty-headed little thing, barely out of school.
Synonyms: scatterbrained, simple, stupid, silly More Synonyms of empty-headed
COBUILD Advanced English Dictionary. Copyright © HarperCollins Publishers
Word Frequency
lacking intelligence or sense; frivolous
Collins English Dictionary. Copyright © HarperCollins Publishers
frivolous and stupid; silly and ignorant
Webster's New World College Dictionary, 4th Edition. Copyright © 2010 by Houghton Mifflin Harcourt. All rights reserved.
Example sentences containing 'empty-headed'
After all, she was not unattractive; a bit empty-headed perhaps, but quite amusing at times.FINAL RESORT (2001) THE GREAT AND SECRET SHOW (2001)
Unlike the beautiful but empty-headed courtesans of some of the aristocracy, Vanozza was a practical woman, which Rodrigo admired.THE FAMILY (2001)
Trends of 'empty-headed'
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Definition of empty-headed from the Collins English Dictionary
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AC/DC: Maximum Rock & Roll: The Ultimate Story of the World's Greatest Rock-and-Roll Bandby Murray Engleheart, Arnaud Durieux
Over three decades and more than 150 million albums, AC/DC has established itself as much more than just a great rock band. For millions of fans spanning several generations across the world, they are an ear-bleedingly loud, sweat-soaked religion, courtesy of such classic albums as Highway to Hell and Back in Black.
Now, in a book of/p>/em>/em>
Over three decades and more than 150 million albums, AC/DC has established itself as much more than just a great rock band. For millions of fans spanning several generations across the world, they are an ear-bleedingly loud, sweat-soaked religion, courtesy of such classic albums as Highway to Hell and Back in Black.
Now, in a book of astonishing breadth and scope, comes, for the very first time, the complete story of AC/DC. Everything you ever wanted to know and plenty more you never dreamt of is all here, the ultimate balls-out adventure, laced with sex, drunken escapades and brawls. It's a journey that started in the suburban Sydney, Australia, bedrooms of brothers Malcolm and Angus Young, boys who could wreak havoc with their guitars. Over their power chords were the lyrics and voice of Bon Scott, who would lead them higher and higher—until his tragic death in 1980. The bittersweet irony after his death was that not only did the Youngs manage to hold together without him, but the band's fortunes and status skyrocketed with his replacement, Brian Johnson, and the album Back in Black.
Five long years in the making, AC/DC: Maximum Rock & Roll is sourced from more than 1,300 interviews the band has given over the past thirty years combined with in excess of 75 of the authors' own interviews with those who worked with AC/DC both in the studio and on the road—many of whom have never spoken about the band publicly. It's topped off with stunning, never-before-seen photos to create the ultimate portrait of the ultimate rock band.
Editorial Reviews
Product Details
- 9780061844539
- Publisher:
- HarperCollins Publishers
- Publication date:
- 10/13/2009
- Sold by:
- Format:
- NOOK Book
- Pages:
- 496
- Sales rank:
- 583,682
- File size:
Read an Excerpt
AC/DC: Maximum Rock & Roll
The Ultimate Story of the World's Greatest Rock-and-Roll Band
Chapter One
With a Glasgow kiss
The crushing mid-1960s summer heat in North Queensland, Australia's last wild frontier and Deep North to America's Deep South, was punishing enough. But the attitude of the locals presented a danger to The Easybeats and their touring partners, The Purple Hearts, that was far greater than heatstroke. No-one liked strangers in these parts, certainly not strangers that looked as they did. Something had to give and in Cairns it did, while Hearts guitarist Barry Lyde, aka Lobby Loyde, waited for a hamburger. An old local who objected to the length of Loyde's hair leapt up and put a knife to his throat. Thankfully, The Easybeats' George Young and Harry Vanda were on hand.
Lobby Loyde: 'George headbutted the prick. He's about two foot tall but, mate, don't get in his road, he's a killer! He just went, "Boof!" and Harry said, "Don't get up!", which he didn't do. Harry could throw a good blow too, I might tell you.'
George's headbutt first, ask questions later approach was an instinctive reaction from his days growing up on the streets of Cranhill, Glasgow.
Cranhill came into being in the '50s when the government decided to forcibly move a mix of people from the tenements of central Glasgow to an area in the east. The housing scheme was based on similar highly successful projects in southern Germany, however on the outskirts of Glasgow it failed spectacularly. The venture became a ticking social time bomb compounded by being out of sight and so out of mind when it came to facilities. By the '60s, razor gangs hademerged on the streets.
It was in this environment that William Young (born 1910) and his wife, Margaret (born 1915), were raising eight children Steven (1933), Margaret (1936), John (1938), Alex (1939), William (1941), George (1946), Malcolm (1953) and Angus (1955).
Steel production and shipbuilding were the city's key industries, but work was hard to come by, a situation that weighed heavily on William, a spray painter who had been a ground mechanic in the air force during World War II.
Music was an ever-present distraction. Margaret, the revered and only sister among the seven brothers, acted as an evangelist of sorts for all manner of blues, jazz such as the glorious freewheeling of Louis Armstrong R&B and the earliest driving rock and roll recordings of Fats Domino, Little Richard and Chuck Berry.
It wasn't long before various members of the family began to play a range of instruments, from guitar, piano and sax to accordion and clarinet. John and Alex picked up the guitar first, while Stevie squeezed the life out of the piano accordion.
In the early '60s, Alex took that grounding into the wider world and secured work in Germany playing saxophone and bass with Tony Sheridan, whose former backing band was a then unknown Beatles.
The decision of Alex to break free with music dramatically increased the daydreams of George, who had shown great promise on the football field. The Youngs were, of course, football fans and supported Glasgow Rangers who, in the '50s, coincidentally boasted a fullback by the name of George Young. In 1961, Rangers became the first Scottish side to reach a European final only to be well beaten, 4-1.
As the '60s progressed, though, the club continued to rack up the trophies on home soil, including winning the Scottish championship and FA Cup in 1964.
By then the Youngs were on the other side of the world, after relocating to Sydney in 1963 on the strength of the Australian Government's £10 immigration package. Maybe in Sydney, Angus, who was eight, could avoid being hit by a car a second time, as he had been in the streets of Cranhill. He made his mark as soon as he arrived, decorating the airport with whatever was in his stomach.
Despite Australia's sun-drenched reputation, it poured rain constantly for six weeks when the family first arrived. To make matters worse, they had to share their basic living quarters in barrack accommodation at the Villawood Migrant Hostel in the city's western suburbs with local snakes and lizards desperate to find somewhere, anywhere that was dry. Everyone was horribly homesick and one night both William and Margaret were reduced to tears. From that moment on, the already strong bonds between the family tightened even further and they were determined to make a success of their move.
William had an additional headache. Apart from the increased employment prospects, he had hoped that the relocation would also act as a circuit breaker on the growing musical career aspirations of some of the family. It was a major if understandable miscalculation. Rock and roll was well established in Australia by the time the Youngs arrived. The switch had been flicked in 1955 with the screening of teen rebellion movie Blackboard Jungle, and its musical soundtrack in the form of Bill Haley and The Comets' Rock Around The Clock transformed normally sedate cinemas into virtual dancehalls. Then came the main players in person.
In January 1957 it was Haley and in October Eddie Cochran, Gene Vincent and Little Richard, while the arrival in Australia of the music of Elvis Presley and a January 1958 tour by Jerry Lee Lewis and Buddy Holly and the Crickets slammed shut all the remaining fire exits.
Not content with a typically manic performance, Little Richard also made history. During the flight to Australia, the deeply spiritual singer felt that the plane was being kept in the air by angels. Then when Russia's Sputnik satellite appeared in the night sky over the Sydney Stadium, he believed the sight was a directive from God. In response, he promptly announced his retirement from rock and roll and his entry into a life of religious service. The huge ring he threw into Sydney Harbour the following day in order to demonstrate the level of his faith is still somewhere in the murk.AC/DC: Maximum Rock & Roll
The Ultimate Story of the World's Greatest Rock-and-Roll Band. Copyright © by Murray Engleheart. Reprinted by permission of HarperCollins Publishers, Inc. All rights reserved. Available now wherever books are sold.
Meet the Author
Murray Engleheart is one of Australia's most respected music writers with a career that spans more than a quarter of a century. He is a noted authority on AC/DC and has interviewed the members of the band on numerous occasions.
Rock journalist Arnaud Durieux has followed AC/DC's career for more than twenty-five years and has amassed the largest archive of AC/DC material in the world. He has interviewed the band many times and has contributed to many of their projects, recently co-producing their multi-platinum DVD "Plug Me In."
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I love AC/DC and rrading this book made me feel like I was a part if their family, from their early years of struggle beire they evwr made it out of Australia through the ups and diwns of managers and replacemnt band mmebers, Bon' untimely death all throughout the rich history that AC/DC has stained in my mind forever* I am eternally grateful fir AC/DC's epic contribution to Rock and Roll* No matter what their influences were AC/DC owned it all!!! Miss you always Bon and LET THERE BE AC/DC FOR LIFE!!!!!!!!
Im minty! She flips her emo hair. And i have better emo hair than Lin over there.
Arent these reveiws suppose to be about acdc not some little kitten clan?
He looks around
"You're welcome", she mewed, then applied the cobwebs gently. J u n e f l o w e r
Where darkstar sleeps


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before the baby is born he is accustomed to live in the womb of his mum.a her out of her stomach must have to him many temp accustomed to the new world he discovers to help it feel security we have to provide a lot of intent.
wear it with his arm support on the sides his arms near his legs like a corp sposition the fetus.
the baby with this new environment does not sleep well it is used are different positions in the womb.
with the free use of the arms and legs that had not before birth do not sleep better aid then must swaddling during all hours of naps during the day or night.
Used in hospitals nationwide, the Halo SleepSack Swaddle is the #1 choice of hospitals and is recognized as "Hip Healthy" by the International Hip Dysplasia Institute. Its strong adjustable fasteners ensure a perfect fit and resist breakouts for more secure swaddling. Plus, its innovative zipper, unzips from the bottom allowing your baby to stay swaddled for easy diaper changes. HALO also proudly supports non-profit organizations like First Candle/SIDS Alliance. Use over regular sleepwear to take the place of loose blankets. Discontinue swaddling when the baby shows signs of rolling over or breaking out of the swaddle wrap. 100% cotton interlock. TOG: 1.5 Size newborn fits 6-12 lbs, size small fits 13-18 lbs.
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Electro Harmonix Hog2 Guitar Synthesizer$ 382.80 $ 478.50
Ludlow Guitars is discontinuing carrying this select brand. And will no longer be Authorized Dealers of these companies. Some of the pedals have been demoed and may show signs of wear. However, all pedals are still under manufactures warranty and are technically brand new with reduced prices. If you want to know the exact condition on a specific pedal, please contact us. Once we sell out of stock that's it, there will be no more. So act now before it's too late.
All Sales are Final
The award winning HOG (Harmonic Octave Generator) earned kudos from players and reviewers alike for its ability to exponentially expand a musician's sound palette. Now EHX take the HOG to an even greater level of performance and functionality with the HOG2. It delivers complete control of ten totally polyphonic and glitch free voices ranging from two octaves below to four above the instrument's pitch—all without the need for a special pickup or instrument mods of any kind.


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Brown healthy and comfortable in role
He's been here a long time. Jeremy brown his going into his senior season, his sixth season as a Gator, and he seems like he has everything in order to do big things when he is done with the year. He has played quite a bit of football in his five years here and he has missed even more with injury. Now, he's going to work and enjoy his final season on campus.
Full Story
March 20, 2013
Brown Excited About Muschamp Hire
It isn't as if the Florida football team was looking for a change at the top, but once some of them found out Will Muschamp was Florida's new head coach and then met him, they have a new vision of the program. Jeremy Brown started most of the season at cornerback for Florida. The sophomore from Orlando has dealt with both the loss of his starting job and now his head coach.
Full Story
December 16, 2010
Muschamp with a Good First Impression
A coaching change is something players might prepare themselves for but you never know how they will handle it until it happens. When Urban Meyer announced his resignation, many players were confused--and rightfully so. But, how are they responding now that the dust has settled and revealed their new coach?
Full Story
December 14, 2010
Lots of Similarities Between Gators and Tide
Jeremy Brown looks at the Alabama football program and feels like he's looking into a mirror. The scheme on both sides of the ball is different, but the talent of the players and coaching staff is why Brown believes it will be the top two programs in college football facing off this Saturday in Tuscaloosa.
Full Story
September 28, 2010
It's a Mental Thing for Brown
The Florida Gators are off to a torrid start in collecting interceptions this season, amassing 12 of them in four games. One guy doing his fair share is someone that never played a down for Florida before this season and has made huge strides physically just to get back on the football field. With all his physical prowess, Jeremy Brown thinks it is the mental part of the game that sets him apart.
Full Story
September 26, 2010
Brown Makes Mistake…Makes Amends
Sophomore Jeremy Brown is trying to make up for lost time. Two years away from the game due to injury, Brown is part of a Florida defense that has flexed its muscle in terms of turnovers. Brown had a big day on Saturday and was able to experience the highs and lows of playing the game he loves, all in one afternoon. The whole time, he continues to live the dream.
Full Story
September 19, 2010
Brown's Quick Rehab Helped the Gators
Friday night was full of uncertainty for Jeremy Brown. After missing the two seasons because of injury, Saturday's opener against Miami (OH) was set to be his first time on the field as a Gator. That was until an ankle injury during Thursday's practice put his chances in jeopardy.
Full Story
September 7, 2010
Brown Fought Through Pain
Florida sophomore cornerback Jeremy Brown fought through the pain of an ankle sprain on Saturday and played in his first collegiate game. Along the way, Brown also corralled his first interception at the collegiate level. Brown has been off the playing field for two long years and his chance to compete was something he looked forward to for a long time.
Full Story
September 4, 2010


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Pacific Northwest National Laboratory, Signatures Science and Technology Division, Richland, Washington, United States
In microbiology research, there is a strong need for next-generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures, such as in bacterial biofilms. White-light interferometry (WLI) can provide high-resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm's interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a nondestructive manner. We build on our prior description of static biofilm imaging and describe the design of a dynamic growth flow cell that enables monitoring of the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is designed to grow biofilms in dynamic conditions and to create a reflective interface on the surface while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope's objective lens. Example images of live biofilm samples are shown to illustrate the ability of the flow cell and WLI instrument to (1) support bacterial growth and biofilm development, (2) image biofilm structure that reflects growth in flow conditions, and (3) monitor biofilm development over time nondestructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). This development will open opportunities for the use of WLI in bioimaging.
Biofilms are communities of microbes that attach to surfaces.1–3 They are ubiquitous in the environment and can have beneficial or deleterious effects on the host structure or organism. Dental plaque is an example of a biofilm that forms in the human body and may cause tooth decay without proper hygiene.4 On the other hand, a biofilm forming at the influent of a sand drinking water filter—the so-called "schmutzdecke"—contributes to removal of harmful bacteria and viruses despite the fact that it may be composed of a natural mixture of bacteria, fungi, protozoa, algae, and even insect larva.5 A characteristic of biofilms is the formation of a protective layer of excreted organic substances known as the extracellular polymeric substance (EPS). The EPS serves to ensure continuous hydration of a biofilm, aids adhesion to surfaces, and often increases the ability to withstand environmental fluxes and treatment with antibiotics or other antiseptic chemicals.6
The EPS layer influences the three-dimensional (3-D) architecture of biofilms, and this structure, in turn, has been shown to influence the severity or harmfulness of the biofilm on the host surface.7 As such, greater understanding of biofilm structure and its role in function may prove valuable in the fight against diseases that are either caused by or exacerbated by the formation of biofilms. It is particularly important to study the early stages of biofilm formation because even a small level of bacterial colonization on a surface can lead to a serious infection on, for example, an implanted medical device. Biofilms are associated with a majority of chronic infections, where they are also increasingly resistant to traditional treatment methods, such as antibiotics.8,9
Studying biofilms in their natural environments is challenging, and nondestructive experimental assay methods are needed.10 It is often desirable to study a biofilm without significantly impacting its normal structure and function because many important biological questions can only be answered by observing a biofilm's transition as a result of an external perturbation. For example, it may be important to identify the location and function of bacterial cells that persist after treatment with antibiotics.11 These cells exhibit enhanced antibiotic resistance and can remain on a surface and repopulate a biofilm even after extended efforts to remove them. In a mature biofilm, cells can detach and spread biofilm infection to new locations. Figure 1 shows the process for biofilm formation and the mechanism for resistance to treatment.
(a) An illustration of the biofilm development process. Biofilms attach to a surface through excretion of EPS. They can then spread and release from surfaces to colonize new areas. (b) Resistance to treatment is mediated by EPS and by the existence of persister cells on the surface, which can repopulate a biofilm.
Optical imaging is a preferred method for nondestructive observation of biofilms. Simple observation with bright-field microscopy can resolve cells and microscopic features. Confocal and fluorescent modalities build on bright-field microscopy with improved resolution, 3-D sectioning, and observation of fluorescent labels. However, each imaging technique shares a common limitation: high resolution and high magnification necessarily limit field of view. Biofilms are spatially heterogeneous, so looking at a narrow field of view may give an incomplete picture of the biofilm's development and may, for example, miss the presence of remnant cells and other biomolecules. Remnant cells may enter a slowed metabolic state (i.e., persister cells), but, even if the cells are dead and merely adherent, they could hasten repopulation of a biofilm even if they are sparsely distributed. Likewise, imaging a large field of view results in poor resolution such that cells and EPS structures cannot be resolved. High-resolution electron microscopy techniques suffer from this limitation, and they typically require destructive sample preparation and exposure to high vacuum. Figure 2 shows a comparison of 3-D imaging techniques that have been applied to biofilms. Atomic force microscopy can provide very fine axial and lateral resolution but requires contact with the sample, which may be disruptive. Optical coherence tomography employs infrared light and has limited resolution as a result. Interferometric imaging [i.e., white-light interferometry (WLI)] is unique in its combination of high axial resolution and large field of view.
Comparison of imaging modalities used to collect 3-D topographical information from biofilms. Typically higher resolution necessarily limits the field of view. The axial resolution of interferometric imaging is independent of magnification, so high-resolution imaging is possible even with a large field of view. This is valuable for imaging biofilms, which are spatially heterogeneous.
WLI offers unique capabilities for biofilm imaging. It is a noncontact, nondestructive optical technique that takes multiple bright-field optical images through a user-determined vertical range and uses interference in the images to recreate a 3-D topological map of the surface of the sample being imaged. In WLI, a broad spectrum light source with low coherence, such as a light-emitting diode (LED), is used to illuminate a sample. Light reflected from the sample is recombined with light reflected from a flat reference mirror to create an interference pattern on a camera sensor. The short coherence length of the light means that interference only occurs over a narrow range when the sample is the same distance as the reference mirror. WLI gained its name from this phenomenon, although, in practice, light of any color may be used as long as it has a short coherence length. Until recently, the application of this technique in microbiology was limited due to the challenges of imaging in liquid media. A biofilm is largely composed of water and typically has a refractive index that is substantially similar to the bulk liquid in which it is grown.12,13 In short, the biofilm surface is not reflective, so it cannot be imaged using WLI, which requires reflection of light from the surface of interest. In some cases, living cells have been imaged through observation of subtle changes in the optical path length (OPL) of light reflected from a reflective substrate on which the cells are supported.14–16 With high axial resolution and a large field of view, WLI is well-suited to investigate the structure and topology of bacterial biofilms.
We recently reported a method for biofilm imaging with WLI that employs a small volume of air to create a reflective interface at the surface of a biofilm.17 The technique uses a fine needle to insert a microbubble between a glass coverslip and the surface of a biofilm. Bubbles are usually deleterious to biofilm studies in flow cells, and many strategies have been devised to remove them.18 However, in this case, the bubble provides a key advantage—it creates a highly reflective interface at the surface of the biofilm that is suitable for a reflection imaging technique, such as WLI. The method was previously demonstrated on biofilms grown under static conditions in petri dishes. In this work, the design of a flow cell suitable for WLI imaging is presented. Flow cells are commonly used for microscopy of biofilms as it is desirable to grow biofilm samples under dynamic conditions that more closely approximate conditions seen in nature.19 They are typically designed with influent and effluent channels leading to and from an imaging chamber. Imaging in flow cells is desirable because the dynamic flow conditions mean the biofilm's growth can be sustained for longer periods of time with a steady supply of nutrient media, and the system is amenable to the introduction of perturbations, chemicals, and therapies through the flow stream. Herein, we describe a flow cell that was designed to enable detailed imaging of biofilm structure as grown in dynamic and well-controlled conditions.
A flow cell for WLI imaging of biofilms was designed with 3-D modeling software (Sketchup). The enclosure for the growth chamber was 3-D printed (Flashforge Dreamer) from acrylonitrile butadiene styrene via a polymer extrusion method. The enclosure was designed to support a substrate and viewing window parallel to one another with influent and effluent liquid channels on opposite ends. Thin glass coverslips (No. 2, 25×50 mm) were used for the viewing window and substrate (Ted Pella). A needle (31 G) was used to deliver liquid to the space between the viewing window and the substrate. Silicone adhesive was used to seal liquid between the two parallel windows. Silicone tubing was used to pump liquid media from a 1-L reservoir to the inlet of the flow cell. Similarly, effluent liquid was pumped to a waste container via silicone tubing. A peristaltic pump (Ismatec REGLO) was used to move liquid through influent and effluent tubing. A wye connection was placed in the tubing just before the inlet with a septum on one branch. The flow cell design will be described in further detail below.
A culture of Pseudomonas fluorescens (ATCC 13525) was grown overnight in liquid nutrient broth (BD Biosciences) on a rotating platform at room temperature. P. fluorescens is a common gram-negative bacterium that is found in soil and water. A 100-μL aliquot of the bacteria was used to inoculate a 1-L bottle of sterile nutrient broth, which served as the reservoir for the flow cell system. Bacteria colonized the imaging chamber naturally, and biofilm was allowed to grow unimpeded for 24 h before imaging. The flow rate was ∼300 mL/day—media was not circulated.
Biofilms were imaged with a white-light interferometric microscope (Bruker Contour Elite GT-I). A 2.5× interferometric objective lens was used; 0.55× and 2× field of view lenses were also utilized. The flow cell was fixed on the microscope's translating stage, and flow was temporarily interrupted during measurement. While the flow cell enables growth of biofilms in dynamic conditions, it is necessary to temporarily pause the flow for imaging because WLI is highly sensitive to sample movement. An air bubble (∼300 μL) was introduced into the imaging chamber of the flow cell through a septum attached to the wye connection just before the inlet. Standard procedures were used to find the focus and level the viewing window of the flow cell. Then, the objective was lowered and scanned through a range designed to capture interference fringes from the lower surface of the viewing window and the biofilm without collecting data from the upper surface of the viewing window or any other surfaces. Raw frame data were saved and analyzed separately from the 3-D profile generated by the instrument software. Bruker's Vision64 software was used for limited data processing and controlling the instrument. Data were also analyzed in both MATLAB® and open source 3-D data visualization software (Gwyddion). At the conclusion of data collection, the flow of liquid media was resumed and the air bubble in the imaging chamber was removed through the effluent channel.
Prior use of WLI to image biofilms was done by placing a clean coverslip over a biofilm and inserting an air bubble under it.17 Light was transmitted through the top viewing glass, but, because it did not have any surface attached bacteria, light could pass through to the biofilm growing on the substrate. At the end of the imaging process, the coverslip was removed, cleaned, and dried before being used for subsequent images. Building an enclosed flow cell offers the advantages of greater control over growth conditions and less impact on the fragile biofilm structures during imaging. However, a significant challenge of using a fixed viewing window that remains in contact with the inoculated liquid is the inevitable accumulation of biofilm on the viewing window. The flow cell was, therefore, designed such that the top viewing window also served as the growth surface for biofilm. For imaging, light passed through the viewing window and the biofilm before being reflected from the lower surface of the biofilm. Compared to the static imaging method, this significantly simplifies calculation of biofilm thickness and reduces the scanning range needed, which also reduces the time needed for measurement. A drawback is that light must pass through the biofilm twice, which limits the technique to biofilms that are partially transparent. It is straightforward to observe the smooth lower edge of the viewing window and measure the distance to the surface of the biofilm. Since WLI is usually performed in air, a scaling factor equal to the refractive index of the medium (i.e., the biofilm) is needed to calculate the thickness.
Figure 3 is a 3-D model of the flow cell used to image biofilms. Its salient details are (1) the large area viewing window, (2) the inlet and outlet tubing that supply nutrient media, and (3) the enclosure that supports the viewing window and maintains a fully enclosed environment for biofilm growth. Latex sheets (McMaster-Carr) of the desired thickness (300 μm) were used to create a gasket, and silicone glue (GE 280) secured the two layers together to create a watertight seal. Tube coupling fittings and sockets (McMaster-Carr) allowed for access and connection to the silicone rubber tubing (McMaster-Carr) that connected the imaging cell to the peristaltic pump. The tubing connected the flow cell to the reservoir of nutrient broth and the waste reservoir.
3-D model of a flow cell used to grow and image biofilms. The viewing window serves as a support for biofilm growth and enables interferometric imaging. The viewing window is 25×50 mm.
Figure 4 gives a more detailed view of the biofilm growth chamber. The viewing window and substrate are parallel to one another and form a thin volume for biofilm growth. This chamber is thick enough (∼300 μm) to support the development of biofilm structures that are much larger than the individual bacteria (1 to 10 μm). However, the space between the viewing window and the substrate was limited to ensure that small bubbles that enter the chamber will be pressed against the biofilm surface. Figure 4 shows that chamber is full of liquid, and biofilm accumulates on the top and bottom surfaces of the chamber when the flow cell is in growth mode. The only change necessary for imaging is the introduction of a bubble, which displaces liquid broth but cannot displace the biofilm, which is more firmly attached to the surface. For initial experiments, air was used to create the bubble, though other gases could be used if anaerobic conditions are desired. Overall, the flow cell enables imaging of biofilms grown in dynamic conditions. The large viewing window makes it possible to observe large-scale biofilm features and heterogeneity in the way that the biofilm forms. The flow cell makes it easy to introduce perturbations to the environment and observe resultant changes in biofilm structure. For example, in future experiments, it will be possible to observe the effects of antibiotics and other antibiofilm therapies by introducing the drugs into the liquid stream. This offers a controlled way to study biofilm persistence.
Cross-sectional view of flow cell shows the thin liquid layer between a glass coverslip and a substrate that forms the growth chamber. Two projected views show the flow cell filled with liquid nutrient broth that supports biofilm growth and with air inserted for imaging. In growth mode, liquid media is pumped through the channel to supply nutrients to bacteria, which form biofilm on all upper and lower surfaces of the flow cell. In imaging mode, an air bubble is introduced into the channel. The air presses against the biofilm, providing a reflective air/water interface that is suitable for imaging. The viewing window is 25-mm wide, and the space for biofilm growth is ∼300 μm.
An important consideration when imaging biofilms in a closed flow system is the selection of the viewing window. Glass coverslips are typically placed over objects for viewing with an optical microscope where their main function is to keep the sample flat and to prevent contact between the immersion liquid (typically oil or water) and the sample. Coverslips are not typically used with WLI imaging because there is no immersion liquid and because the smooth glass would obscure the texture of the underlying sample. For biofilm imaging with WLI, the coverslip is integrated into a flow cell and serves to contain the liquid media that supports bacterial growth. Bacterial adhesion is nonspecific and forms on all interior surfaces of the flow cell, including the bottom of the coverslip. The coverslip serves as the growth surface for biofilm that is to be imaged and as a viewing window. As with any interferometric imaging through transmissive media, it is important that the coverglass is transparent and smooth. The thickness of coverslip glass is a limiting factor when imaging biofilms. Light transmitted through the higher index coverslip glass traverses a longer OPL than it would through air. The increase in OPL from the coverslip cannot exceed the depth of field of the objective (∼133 μm for experiments in this study). If the shift in OPL is greater than the depth of field enabled by the objective lens, then the image will not appear in focus at the length fixed in the reference arm of the interferometer. However, calculations shown below reveal conditions in which transmitting light through a thin coverglass over the sample will not prevent light interference.
Using Eq. (1), the depth of field (dtot) for an interferometric objective can be calculated. For experiments in this work, the lens had 2.5× magnification (M) with numerical aperture (NA) of 0.07. The middle wavelength (λo) of illuminating light was ∼550 nm (from a green LED). The smallest lateral resolving distance (e) is 3.8 μm, and the refractive index (n) of the immersion media (air) is 1. Given these parameters, the depth of field is ∼133 μm, which is in the order of the thickness of the cover glass. Since the angle of incidence is small, the OPL through glass can be approximated as the thickness multiplied by the refractive index (OPL=T×n), and the optical path difference (OPD) is simply the difference between the thickness and the OPL (OPD=OPL−T). Table 1 shows the thicknesses of various grades of glass coverslips and the corresponding OPD caused by imaging through the glass Display Formula
Coverslips up to ∼250 μm are suitable for transmission imaging with a low-magnification lens because the OPD does not exceed the depth of field. Table 2 shows the decline in depth of field at higher magnification. The narrow depth of field for high-magnification lenses limits the ability to image through glass viewing windows. Even at magnification as low as 5×, the depth of field is prohibitively narrow. With higher magnification it would be necessary to compensate for the OPD by inserting a sample of equal thickness in the reference arm of the interferometer.20 In any case, the spatial resolution of the low-magnification lens is sufficient for observing a biofilm's morphology, and the large field of view is actually an advantage over other imaging modalities. The thickness of the biofilm must also be considered when selecting a viewing window, as the water contained in the biofilm will also contribute to a shift in OPL. For experiments in this work, No. 2 coverslips were used because their added thickness reduced the incidence of cracking as a result of pressure induced by the pumping system. Thinner viewing windows would be desirable for imaging thick biofilms but may also require added support because of the tendency to crack under the pressure induced by pumping nutrient broth through the system.
Imaging through a glass coverslip results in multiple sets of interference fringes. Figure 5 shows that interference is seen at the top surface of the glass window and at the bottom surface of the window, which is where it meets biofilm. A final set of fringes with lower intensity is observed at the surface of the biofilm when an air bubble is present. Although the amplitude of this latter fringe packet is much lower than that of the prior two, it is still easily measurable above the noise floor. The refractive index of the glass and the biofilm must be taken into account when calculating the physical distance between each surface. In practice, a vertical range that starts just below the first air/glass interface is selected when collecting data, which results in capture of only the final two sets of interference fringes. Together they can be used to measure thickness and surface roughness of the biofilm. Imaging artifacts can be observed if the biofilm is too thin because the interference fringes overlap and cannot be easily deconvoluted.
Imaging through a glass coverslip results in multiple sets of interference fringes. The figure shows an example of multiple interference fringes observed at a single pixel. Interference is observed at the top and bottom of the glass viewing window and from the surface of the biofilm. Light is transmitted through the glass and the biofilm to reach its surface. Each frame on the horizontal scale is a two-dimensional image that is collected during vertical scanning. The physical distance between each surface can be calculated using the group refractive index of each medium (air, glass, and biofilm).
Figures 67–8 show examples of WLI images of biofilms grown under flow conditions in the flow cell described above. These examples were chosen to demonstrate the strengths and limitations of using WLI to image biofilms. Figure 6 shows the complex heterogeneous structure of a bacterial biofilm on the left side of the image. There are small independent colonies and larger interconnected structures. The right side of the image is smooth, but not because biofilm is absent in this region. The arc that cuts through the center of the image is the leading edge of the air bubble that was inserted into the flow cell. Biofilm is visible and can be imaged interferometrically with the air bubble but cannot be seen without the air bubble.
Example of biofilm image at the edge of a microbubble of air as captured in a flow cell. The left side of the image shows the interconnected nature of a developing biofilm of P. fluorescens. Biofilm is not visible at the right of the image because the air bubble was not present under this area. The edge of the bubble is indicated with an arrow. The image is false colored according to the vertical scale at right. The horizontal scale shown by the bar is 500 μm.
WLI image shows a flow channel among small scalloped microfeatures of a P. fluorescens biofilm. The presence of the channels and the morphology of biofilm features are indicative of growth in flow conditions. Flow of liquid media was from left to right. Features can extend over millimeters or result in small separated microcolonies. The image is false colored according to the vertical scale at right. The horizontal scale shown by the bar is 500 μm.
A section of the image shown in Fig. 7 is shown here in 3-D. WLI produces high-resolution topographical images of biofilm. This image is representative of the heterogeneity encountered in biofilms. There is a wide range of thicknesses. The image is false colored according to the vertical scale at right.
Figure 7 also has a region in the center of the image with no biofilm. In contrast to Fig. 6, this is not because the region lacks an air bubble. Here, the hydrodynamic effects of flowing liquid media have created a channel where biofilm growth was limited. This image highlights one of the strengths of WLI imaging. The large field of view ensures that a large and salient feature is not overlooked. Imaging with a traditional method with a more limited field of view would require stitched assembly of a large number of images to attain the same level of detail. The biofilm structures seen in Fig. 7 also indicate hydrodynamic influence. In the image, flow of liquid media was from left to right. The biofilm has formed small scalloped features as a result of liquid flow. Figure 8 shows a portion of Fig. 7 in 3-D. The images indicate that the combination of WLI imaging and flow cells could be used to study hydrodynamic effects on biofilms.
As described above, WLI is a noncontact and nondestructive imaging method. However, it is clear that the flow cell and air bubble used to create a reflective surface require physical contact with the biofilm and this may impact the structure of the biofilm, especially where bacteria are weakly attached. The interface of a biofilm is known to contain streamers of loosely attached bacteria. These features may be removed or compressed with the insertion of air. In a prior study of the effects of shear flows on biofilms, it was shown that wall shear stress can be calculated if the hydraulic diameter and flow velocity of the system are known.21 Hydraulic diameter (Dh) is calculated from the height (H) and width (W) of the flow channel Display Formula
The highest flow velocity encountered in the system can be estimated from the average flow velocity (vave) Display Formula
Then the wall shear stress (τw) can be calculated from the hydraulic diameter, the maximum flow velocity, and the dynamic viscosity of water (η) with the following equation: Display Formula
In this study, the average flow velocity was estimated to be 4.5 mm/s (the bubble filled a 45-mm-long channel in 10 s) with maximum flow velocity of 6.725. The hydraulic diameter was 0.8 mm (H=0.5 mm and W=19 mm). A reference value of 0.001 Pa·s was used for dynamic viscosity of water. Then the shear stress on the biofilm was estimated to be 0.034 Pa. The aforementioned study of shear stress on biofilms21 indicates that this shear stress is in line with the shear stress encountered during low-velocity growth phase (0.04 Pa in that study) and at least 1 order of magnitude lower than shear stresses that caused irreversible compression and/or erosion of the biofilm (0.6 to 3.6 Pa). The advantage in the method presented here is that the bubble can be introduced at a very low velocity to minimize impact. Our experiments showed that the bubble did not remove biofilm completely, although further study is needed to study the impact of the imaging process.
In addition to shear stress, the bubble presses down on the biofilm and conforms to its surface. In practice, the conformation of the bubble exerts a normal pressure that is not significantly higher than atmospheric pressure. Nonetheless, it is still higher than the pressure exerted when no bubble is present. Thus, it is likely that the biofilm is compressed when the bubble is present, especially with components with less structural integrity. A comparison of images collected with WLI and a traditional optical technique, such as confocal microscopy, would be useful for confirming this and quantifying the effect.
The results presented in this paper demonstrate that WLI is suitable for biofilm imaging when coupled with an imaging cell with appropriate optical properties. With the flow cell described here, a biofilm can be grown in dynamic flow conditions and can be imaged at regular intervals. Nonetheless, there are several limitations that could be overcome in future research. Multiple measurements have been made in the same location, but more research is needed to establish repeatability of the method. This would also better characterize any adverse effects of placing an air bubble in direct contact with the biofilm. To date, multiple measurements of a biofilm indicate that larger colonies and aggregate structures are persistent; however, lateral resolution of the optical microscope at low magnification is too low to see effects on individual microbes. Finally, while the flow cell described in the paper enables growth of biofilm in dynamic conditions, it is still currently necessary to stop flow for imaging. In future experiments, it may be desirable to explore the benefits and drawbacks of dynamic laser interferometric imaging systems.
This paper presents the design of a flow cell that supports growth of bacterial biofilm and is compatible with WLI imaging and describes a WLI biofilm imaging methodology. An important feature of the flow cell is the viewing window, which is selected to enable light interference without shifting the biofilm's surface out of the field of view of the objective lens. The viewing window enables WLI imaging of biofilm grown in an enclosed and well-controlled environment. The flow cell has influent and effluent channels that support a consistent supply of nutrient broth to a growth chamber where biofilm is cultured. The flow cell is designed to create an interface at the surface of the biofilm that can reflect light back to the imaging objective. Imaging through a glass coverslip results in multiple sets of interference fringes, and data collected during an extended vertical scanning range can resolve each of these interfaces. Biofilm thickness and surface texture can be resolved in situ and nondestructively. Example images demonstrate that WLI imaging can be used to observe hydrodynamic effects that are indicative of biofilm growth in flowing liquid. This technique will enable future research on the structure, development, and response of bacterial biofilms in well-controlled growth conditions. Given the unique capabilities, further exploration of bioimaging applications for WLI is warranted.
This work was performed at the Pacific Northwest National Laboratories (PNNL), a multiprogram national laboratory operated for the United States Department of Energy by Battelle Memorial Institute under Contract No. DE AC06-76RLO 1830. The work was supported by the Chemical Imaging Initiative and by the Microbiomes in Transition Initiative as part of PNNL's Laboratory Directed Research and Development program. The authors state no conflicts of interest.
Curtis Larimer is a materials scientist in the Chemical and Biological Signature Science group at Pacific Northwest National Laboratory (PNNL). He is engaged in basic and applied research at the nexus of chemistry, materials, and biology. His work has focused on nondestructive chemical and biological imaging.
Michelle Brann was a visiting researcher at PNNL from 2015 to 2016. She is currently pursuing her PhD in chemistry at the University of Chicago.
Jonathan D. Suter is a biomedical engineer in the Applied Optics group at PNNL. He is involved in research aimed at metrology of complex surfaces, including metals, plastics, fabrics, biological tissues, and radiological samples, using both atomic force microscopy and white-light interferometry.
R. Shane Addleman is a senior research scientist at PNNL. His research has focused on analytical methods for chemical, biological, radiological, nuclear, and explosive trace materials. His current work focuses on enhanced trace collection and detection, applied optical and spectroscopic methods, and innovative inorganic and organic separation techniques.
Curtis Larimer ; Michelle Brann ; Jonathan D. Suter and R. Shane Addleman"Design of a dynamic biofilm imaging cell for white-light interferometric microscopy", Opt. Eng. 56(11), 111708 (May 10, 2017). ; http://dx.doi.org/10.1117/1.OE.56.11.111708
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|Reviews for I Come Here Every Night
| Magenta Ann chapter 1 . 12/4/2009
I like this a lot. I'm a huge Cabaret fan, especially of the Emcee. Gotta love a creeper. I hate most of the Cabaret fics I've seen... They always glorify the whole dealio and take away all the perfect sleaziness of the Klub. They always make the Emcee shockingly cutesy and just... Not Emcee-like. But this is realy good, and I don't usually like fics with original characters. You should keep going with this story! I'm thinking about making my own Cabaret fic now, too. :)
| Zafiro girl chapter 1 . 11/2/2009
Although I don't speak english very well, I like to writte a review to say you that I like your fic, for his beautiful story, although I don't like the fics with "self-insert"
| JenniferJoy26 chapter 1 . 4/8/2008
I loved the story. Is this the end of it? please keep writing.