7/15/2023 0 Comments Whiskey a go go new york![]() ![]() The Courtney Villas subbed for the Chateau Marmont in “Daisy Jones and the Six.” “We knew we wanted to shoot on the actual Sunset Strip,” Kender says, “So that was one of the first things locations got moving on, because it’s so hard to shut down.” The crew basically took over the Viper Room block, “so whenever you see Sunset, you’re either looking outside the Whisky or on that one block where the Viper Room is.” The solution? “We did find a softcore porn that we used,” she reveals - complete with a catchy Filthy’s song. While there was enough documentation of the heyday of the Sunset Strip to compile a large reference binder, it was nearly impossible to find photos or film that actually showed the interior when it was still Filthy McNasty’s. River Phoenix tragically overdosed on the sidewalk in front in 1993. In the early 1990s, the bar, then called The Central, was sold to a group of young Hollywood types including Johnny Depp, who renamed it the Viper Room. It’s only natural that the Six would get one of their first gigs at this lower-key spot, which was frequented by the likes of Elvis, Mick Jagger and Jim Morrison. Not quite in the same league as the Troubadour and Whiskey a Go Go, the smaller club Filthy McNasty’s was nonetheless a fixture of the 1970s and ’80s Sunset Strip. The Six gets ready to play Filthy McNasty’s on the Sunset Strip. ![]() The club needed to quickly return to its live music schedule after the art department dressed it for the ’70s, so “we were given like 24 hours, we would run crews morning and night,” she says. ![]() It used to be covered in vintage wallpaper,” Kender explains. “The Whisky doesn’t have go-go cages anymore. But of course, it doesn’t look exactly the same 50 years later. The Turtles, Love, the Byrds and Van Morrison’s Them were just a few of the dozens of other bands whose careers were juiced by Whisky appearances. Jim Morrison got around to just about everywhere in Hollywood, but the Whisky is where the Doors were the house band, and one of the first places where go-go girls frugging and watusi-ing in cages above the dancefloor became popular. music world - the rock club was basically the epicenter of the Sunset Strip scene from the mid-1960s until well into the 1970s and beyond. It’s not surprising Daisy would show up at the Whisky on one of her very first forays into the L.A. “I felt like if we couldn’t get the actual thing, since Duke’s no longer exists, as long as we emulated the feel of the places, that was enough,” Kender says. However, a neon sign for Duke’s was added. Recently reborn as Clark Street Diner, it looks convincingly like someplace Daisy might have slung hash before hitting it big, with its flagstone wall, golden Naugahyde booths and rows of photos of famous patrons. Since Duke’s is long gone, the series shot at the Hollywood Hills Coffee Shop. In those days, it was far more likely to find strummers and songbirds at Duke’s Tropicana Coffee Shop or Barney’s Beanery. The Hollywood Hills Coffee Shop quickly became a Hollywood classic after it got cleaned up with a makeover and new chef in 1994 and was then immortalized in the 1996 movie “Swingers.” Was it a popular hangout for rockers and hangers-on way back in the 1970s? Not really, though it did exist. Riley Keough (Daisy) at the Hollywood Hills Coffee Shop/ Duke’s Lacey Terrell/Prime Video ![]()
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7/15/2023 0 Comments Used hyundai santa cruz truck![]() At just $24440 MSRP you get the plentiful tech, stylish look, entertainment options, and all the lovely benefits. You get all the colossal benefits at an incredibly aggressive cost. The used Hyundai Santa Cruz gives you undoubtedly incredible value at just $24440 MSRP new. Used Hyundai Santa Cruz Tampa Priceįor most clients, choosing what Crew Cab to purchase all comes down to price. So what do Tampa consumers have to say about the used Hyundai Santa Cruz models and find the deal that is best for you. ![]() Since Asbury Auto is one of the nation's biggest automotive groups, our wonderful team gets to work directly with consumers daily from all over the country and hear definitely what clients like and do not like about certain vehicles. Used Hyundai Santa Cruz for Sale Near Tampa We offer tremendously efficient automotive repair and maintenance services for all makes and models as well as parts and accessories that would look great on your used Hyundai Santa Cruz. We believe value is created during the ownership experience, and that comes with selling a quality Crew Cab that has been sold at a reasonable price so you continue to do business with our amazing dealerships. At Asbury, our objective is to price all of our used cars fairly and correctly to sell as immediately as potential. ![]() To make things more complex, new and used vehicle prices can still be impacted more laboriously by geographic location, interest rates, OEM incentives, and other market conditions. There are clearly pros and cons to owning both new and used, and everyone's situation is different. Vehicle depreciation is one of the principal reasons a lot of consumers like to buy used cars instead of new. Shop for one today at Asbury Auto! Hyundai Santa Cruz Tampa Price Used Old school and new school car lovers can come together and love everything the remarkable used Hyundai Santa Cruz provides. Take time to shop everything Asbury Auto has to offer and explore why so many customers pick to shop with Asbury. With Asbury Automotive's powerful reach, we offer an definitely enormous inventory of new and used vehicles located coast to coast throughout the United States. It has immensely upped the game for the competition and marks a serious leap forward. The used Hyundai Santa Cruz is one of the most tremendously esteemed vehicles you will find on the new or pre-owned market today. To determine the recall status of a vehicle, visit. RECALL NOTICE: Some vehicles offered for sale may be subject to unrepaired manufacturer safety recalls. You can upload important documents such as your driver's license, insurance, and more to save time when visiting any of our dealerships. Clicklane still offers you the ability to completely reserve your vehicle and enjoy the full in-store experience if you select to buy in person. Not interested in financing your used Hyundai Santa Cruz? No problem! Clicklane offers leasing options through most captive lenders and many major banks or you can simply pay in full using our secure online payments platform. ![]() If you are interested in the elegant Hyundai new or used Crew Cab, Asbury Automotive precisely offers the best online vehicle shopping service imaginable using Clicklane, the world's best online loan marketplace where you can select your lender and your rate in a completely clear mobile-friendly environment. ![]() 7/15/2023 0 Comments Will byers season 1![]() ![]() After his days of hiding out, Will was finally captured by the monster and taken to the Upside Down version of the town's library, where the monster trapped Will in a large, fleshy membrane, littered with skeletons and corpses. RescuedĮventually, Will's friends and family figured out how to locate and save him, and it was just in the nick of time. He listened to his favorite song by The Clash on a mixtape made for him by Jonathan, and was even able to briefly speak with a new psychokinetic ally named Eleven when she entered a deep psychic state, telling her to hurry because the toxic air in the Upside Down was making him weak and vulnerable. Will managed to hide from the monster for about a week by retreating to a fort in the dimension, which resembled the fort he had built in the woods near his house in his home dimension, called Castle Byers. He was able to let his mother know he was still alive, but was in danger. He managed to evade the monster and even learned how to manipulate electricity to communicate with his mother via a system of holiday lights. While in the monster's home dimension, which Will's friends came to refer to as the Upside Down, Will adapted surprisingly quickly. ![]() Will ran to the shed and loaded a shotgun to defend himself, but the monster appeared behind him and took him into an alternate dimension. He rushed to his house but found nobody home - both his mother Joyce and his older brother Jonathan were at work. After he left a Dungeons and Dragons game at his friend Mike's house, Will caught a glimpse of a terrifying monster on his path home. TakenĪlmost immediately after the series began, Will went mysteriously missing. To get ready for Season 2, here's everything you need to know about what Will Byers went through in the first season. ![]() But although Will was out of sight in the first season, he was still the force that brought all the show's main characters together and helped them discover the supernatural occurrences in Hawkins, Indiana. A few months ago, the show's executive producer Shawn Levy confirmed Will is a major player in Stranger Things Season 2, unlike his almost total absence from Season 1. But that's going to change in the upcoming Season 2, which will see Will back from his terrifying vacation in the Upside Down and at the center of all the action. Although the first season of Stranger Things was all about Will Byers, he actually wasn't in it that much. ![]() 7/15/2023 0 Comments Pgsharp key![]() It has a built-in anti-ban system that helps protect your account from being banned by game developers. Safe to use: PGSharp is considered one of the safest spoofing apps for Pokemon Go.You can change the speed of the movement, the frequency of the GPS signal, and many other settings. Customizable settings: PGSharp allows you to customize the settings to your liking.This feature ensures that your location is more precise, which can help you find rare Pokemon. Enhanced GPS accuracy: PGSharp has a feature that enhances your GPS accuracy, which is beneficial in Pokemon Go.This makes it easier to navigate through the game and catch Pokemon. ![]()
7/15/2023 0 Comments Photosmart 5514 printer![]() If you're looking for your printer's ink cartridge, CompAndSave offers a wide variety of cheap ink cartridges that will work on your printer covered with a 1-year satisfaction guarantee. For further information, just click What ink does my printer use? and you will be redirected to the right page. It is also compatible with the high-yield ink cartridges HP 564XL in the same color variation. For example, HP Photosmart 5514 printer ink cartridges are the standard yield HP 564 black, cyan, magenta, and yellow ink cartridges. It is usually a set of numbers or a combination of alphanumeric characters. This model number is used to distinguish the type of ink you should purchase. In front of it is a label or a sticker with the model number. You may also check the existing ink cartridge that is in your printer. To know what type of ink your printer uses, check the manual that came with the printer. Most Common HP PhotoSmart 5514 Ink Cartridges Questions What ink does HP Photosmart use? HP Photosmart has series of printer models that do not use the same ink. ![]() For further questions or inquiries, feel free to contact us, and our Customer Support Representatives will be more than happy to assist you. NOTE: Please be informed that using CompAndSave's ink for PhotoSmart 5514 will never void your printer warranty. Shopping for cheap replacement ink cartridges can't get any better than this! Plus, our low-priced HP PhotoSmart 5514 printer ink cartridges are backed up with a 1-year 100% satisfaction guarantee.Enjoy our free shipping offer for orders over $50 (for orders shipped to all 50 states in the USA).Worried that our printer cartridges might expire before you use them all up? We assure you that our HP 5514 ink cartridges are fresh from the factory every month and have a tested shelf-life of at least 20 months.Make sure to explore different quantities to find the price break for HP PhotoSmart 5514 ink cartridges that suits you best. Get more for less! Enjoy even bigger discounts if you order HP 5514 cartridges in bulk.Not sure about how to install a replacement, remanufactured, or compatible ink cartridges? Worry no more as CompAndSave ensured that installing HP 5514 ink cartridges will be the same as installing the original ones to save you from the hassle.Our replacement HP PhotoSmart 5514 ink cartridges are designed to ensure that you will get the same number of sheets and print quality as the original at a much lesser price.Why choose CompAndSave in shopping for HP PhotoSmart 5514 cartridges? We specialize in providing cheap alternative HP 5514 ink cartridges. Y'all ain't telling me that I am smarter than you, are you? If I can do it, anybody can! Just follow the instructions that the software gives you and you are on your way to using one of the best printer, copier, scanners that I have ever used.CompAndSave is proud to offer wallet-friendly prices on our high-quality replacement HP PhotoSmart 5514 ink cartridges. I am a 64 year old male from Mississippi with limited computer skills and I did it without a hitch. Your computer on USB AND everybody else on the router. ![]() You CAN have a wireless AND a USB connection at the same time. ![]() lala33, this is where it will let you set up your USB connection. ![]() Once that is done, put the CD into your computer and, again, do what it tells you to do. DO NOT connect the USB cable yet!! Do what the printer tells you to do. You get an "It cannot be that easy' feeling and make it harder than it is. Why? Because installation is all automatic and easier than it looks. Installation looks like a nightmare if you see only 3 pages of instructions. The only fault with this printer is the lack of documentation. ![]() ![]() ![]() ![]() 3.7v Battery for Amazon Kindle Fire HD 8 7th Gen Sx034qt Year 2017 Mc-31a0b8. MoKo 7-8 Inch Kids Tablet Sleeve Portable Neoprene Carrying Case Pony Pink. OtterBox Apple iPad (9th gen, 8th gen, 7th gen) Defender Series Pro Case - Black. While the processors and power brought to bear by each Fire tablet do vary, the fact is that none of the models we've listed should feel sluggish, since they're all relatively recent launches. Black Neoprene Tablet Sleeve Case Pouch Carry Bag for 8' Amazon Fire HD 8 Plus. Updated January 2023: Weve updated the Fire HD 8, Fire HD 8 Plus, and Fire HD 8 Kids tablets to the latest models, and added a new section on useful Fire HD accessories. Shop Target for Cases & Sleeves you will love at great low prices. While it doesn't have a screen protector, it. This DJ&RPPQ case is fashionable, rugged, and suitable for the 11th-generation Fire HD 10 tablet. There's no wrong answer here, but anything will narrow it down. The 7 th Generation version can use cards up to 256 GB in size, while the 8 th Generation can manage cards that go up to an impressive 400 GB. DJ&RPPQ Armor Series Full Cover For Fire HD 10. Magnetic connection keeps the cover securely closed. Use vertically to read books or horizontally to watch movies. 32 at Amazon 33 at Best Buy Grifobes Premium for Fire HD 8 (2022) Premium pick Just because the Fire HD 8's cheap doesn't mean it has to look that way. Slim design with built-in stand for hands-free viewing. If, instead, you imagine the tablet staying in your bag for when you're on the move, something smaller is probably a better idea, so that you can more easily bring it with you wherever you go. Description Fits 9th Generation Fire 7 Tablet. This can really help you figure out the right size and model for you - what do you plan to do on your tablet? Is it primarily going to be a media machine for lounging on the sofa or in bed?ĭo you plan to use a bunch of apps and games, or even try to work on the tablet every so often? If so, a bigger display will be handy. ![]() Of course, figuring this out leads to my next question. ![]() The Judiciary said such acts were unfair to both parties and also affected public confidence in the courts.Knowing roughly what size you think you want will help you a lot - do you imagine holding the tablet in one hand, or are you thinking more of a comparison to the standard iPad, in which case a bigger option makes sense? 1 1 result Amazon Fire 7 Tablet Case - Compatible with 9th Generation - Charcoal Amazon 21 24. A retrial under a different judge was also ordered. (105) 54.95 New 24.62 Used Amazon Fire HD 8 (7th Generation) B01J94TG3E 8' Tablet 16GB WiFi - Punch Red (28) 54.95 New 35. The national security judge had already received a serious reprimand from Chief Justice Andrew Cheung after the Court of Appeal ruled that he had plagiarised the plaintiff's submission in his ruling on a trademark case. The Court of Appeal said it will make a judgment after considering relevant evidence on its own, and would give no weight or very little weight to Chan's judgement. The appellant Lo Kai-shui questioned if Chan had independent thinking when handling the case, since he said the judge had incorporated extensively from his brother Lo Ka-shui's submissions in his ruling. The Court of Appeal on Wednesday heard a civil case between two sons of the late co-founder of real estate firm Great Eagle Holdings, Lo Ying-shek. All Generation KPW 3/4th Gen case leather Fire HD 8/10/7 Plus case,Kindle Oasis (3rd)2ndcover Crossbody strap Full Grain Dark Green. High Court judge Wilson Chan is facing another accusation of making extensive use of other people's words in his rulings, after receiving a serious reprimand from Hong Kong's top judge over copying in a separate case. ![]() 7/15/2023 0 Comments Us passport renewal fee 2019![]() ![]() ![]() It cannot be used to travel by air to these countries, or by any means to other countries. It is also accepted for entry by land or sea into Canada, Mexico, and some countries and territories in the Caribbean. passport cards can be used to enter the United States at land border crossings and sea ports of entry. To prevent the RFID chip from being read when the card is not being used, the passport card comes with a sleeve designed to block RFID while inside. ![]() While a biometric passport contains a chip containing all of the traveler's information in electronic format, the RFID chip in a passport card does not contain any personal information beyond the identifying number, which is used to locate records in secure government databases. Unlike the passport book, the RFID chip in the passport card is designed to be readable at a greater distance, allowing border agents to access traveler information before they pull up to the inspection station. In an effort to improve efficiency at land crossings, the passport card also includes a vicinity-read radio frequency identification chip with a unique identifying number tied to government databases. In order to offer a less expensive and more portable alternative to border communities and frequent travelers, the passport card was developed. citizens traveling to Canada would have been required to obtain a passport booklet in order to enter the United States. Previous to the WHTI, many different types of documents were acceptable to cross the border, including birth certificates issued by thousands of different authorities within the United States and Canada. One result of this was the Western Hemisphere Travel Initiative (WHTI), which mandates as of 2007 that a smaller, more secure number of documents verifying both identity and citizenship be used to facilitate identification and international border crossing. National identity cards with similar utility are common inside the European Union and European Free Trade Association countries for both national and international use, with the difference that such cards can also be used for international air travel (within the EU, the Schengen Area and several other European countries that allow entry with a national ID card).Īs a result of the September 11, 2001, attacks, the United States began implementing a range of measures to increase the security of its borders and its identity documents. The card is manufactured by L-1 Identity Solutions. By the end of 2016, more than 12 million passport cards had been issued to U.S. Applications have been accepted since Februproduction of the cards began on July 14, 2008. The passport card (previously known as the People Access Security Service Card or PASS Card) was created as a result of the Western Hemisphere Travel Initiative, which imposed more stringent documentary requirements on travelers. However, the passport card cannot be used for international air travel. The passport card allows its holders to travel by domestic air flights within the U.S., and to travel by land and sea within North America. Department of State, compliant to the standards for identity documents set by the REAL ID Act, and can be used as proof of U.S. passport book, the passport card is only issued to U.S. federal government in the size of a credit card. The United States passport card is an optional national identity card and a travel document issued by the U.S. ![]() Only one administration fee is charged for an individual applying for both a card and book at the same time.) $35 administration fee (required for all minor's cards and passports and an adult's first passport. The rear of a United States passport card (2008)ġ0 years after date of issue for people 16 or older 5 years for minors under 16 ![]() 7/15/2023 0 Comments Decode json string python![]() Get hostname, domain, and protocol from a URL Get the filename and the file extension from a URL Remove all non-alphanumeric characters from a stringĬonvert a character to a code point and vice versa Remove one or many substrings from a string The modern Python regular expressions cheat sheetĬapitalize the first letter of each word in a stringĬompare 2 strings ignoring case sensitivity Generating a random float between min and maxįormat large numbers with comma separators Generate a random integer between min and max If you claim to accept JSON matching a schema, you should accept all JSON matching that schema, otherwise you aren't accepting JSON.Check if a string can be converted to a number To be clear, "as strictly as you can" means respecting the protocol/file format. ![]() See A Patch for Postel's Robustness Principle.) I recommend the LangSec site generally for more discussion of these matters. (If this seems to conflict with Postel's Principle, often sloganized as "Be conservative in what you do, and liberal in what you accept from others", that's because it is commonly misunderstood and the misunderstood form is terrible advice. Generally speaking, you want to parse into a structured representation as soon as you can and as strictly as you can. I assume that isn't the case, because otherwise you'd have no reason to expect it to be JSON at all. If the string is truly an opaque blob with respect to your code as well as to any 3rd party services you pass it to, then you shouldn't be touching it at all. In general, anything you can do to reduce the number of representations for the same value is a good thing for correctness, ease of testing, and security. If the input to that JSON parser is only ever the output of json.dumps, then you only need to consider the output json.dumps can produce, and not all possible legal JSON strings. For example, let's say the JSON parser used elsewhere in your code does have some vulnerability when exposed to legal but carefully formatted JSON. json.dumps) so that you know the JSON is in a consistent, well-behaved format. If you still want to pass it around as a string though, it would be better to reserialize (i.e. You can convert it to a string when you need it as a string. (Ideally, you would also check the JSON against a schema.) Even better, would be to parse into a JSON object and then pass that around rather than a string. This code will then ensure that you are passing valid JSON to those deeper parts of the code. However, to that end, this code is likely slightly better than passing the string through untouched assuming some other part of your system processes it as JSON. This is usually security relevant when one of those pieces of code is doing some kind of security check. ![]() While it doesn't seem relevant here, you can sometimes have security issues where different pieces of code interpret the same input in different ways. I didn't find any CVEs related to json.loads.Īs a potental denial-of-service vector, I haven't looked at the code, but there's no real reason to expect it to be super-linear in either time or space with respect to the length of the string input. This Stack Overflow answer goes into detail for the actual implementation at that time. ![]() Short of bugs in the implementation or monkey-patching, there's no reason it would or should allow executing of anything other than the JSON parsing code. ![]() 7/15/2023 0 Comments Clemens schick shaved head![]() ![]() 238000006011 modification reaction Methods 0.000 description 2.238000006073 displacement reaction Methods 0.000 abstract description 20.210000003491 Skin Anatomy 0.000 claims abstract description 52.239000000969 carrier Substances 0.000 claims abstract description 82.Assigned to AMERICAN SAFETY RAZOR COMPANY reassignment AMERICAN SAFETY RAZOR COMPANY TERMINATION OF SECURITY INTEREST IN PATENTS Assignors: FIRST NATIONAL BANK OF BOSTON, THE, AS AGENT Anticipated expiration legal-status Critical Status Expired - Lifetime legal-status Critical Current Links Assignors: AMERICAN SAFETY RAZOR COMPANY, A DE CORP. Assignors: BANKERS TRUST COMPANY, AS AGENT Assigned to FIRST NATIONAL BANK OF BOSTON, THE reassignment FIRST NATIONAL BANK OF BOSTON, THE SECURITY INTEREST (SEE DOCUMENT FOR DETAILS). RECORDED AT REEL 4324, FRAMES 601 AND REEL 4888, FRAMES 485. ![]() ![]() Assignors: AMERICAN SAFETY RAZOR COMPANY Assigned to AMERICAN SAFETY RAZOR COMPANY reassignment AMERICAN SAFETY RAZOR COMPANY RELEASED BY SECURED PARTY (SEE DOCUMENT FOR DETAILS). Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.) Filing date Publication date Application filed by American Safety Razor Co filed Critical American Safety Razor Co Priority to US06/056,370 priority Critical patent/US4241500A/en Application granted granted Critical Publication of US4241500A publication Critical patent/US4241500A/en Assigned to BANKERS TRUST COMPANY A NEW YORK BANKING CORPORATION reassignment BANKERS TRUST COMPANY A NEW YORK BANKING CORPORATION SECURITY INTEREST (SEE DOCUMENT FOR DETAILS). Original Assignee American Safety Razor Co Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.) Iten Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) Expired - Lifetime Application number US06/056,370 Inventor Clemens A. ![]() Google Patents Razor with adjustable blade positioningĭownload PDF Info Publication number US4241500A US4241500A US06/056,370 US5637079A US4241500A US 4241500 A US4241500 A US 4241500A US 5637079 A US5637079 A US 5637079A US 4241500 A US4241500 A US 4241500A Authority US United States Prior art keywords razor blade substantially circular accordance shaving Prior art date Legal status (The legal status is an assumption and is not a legal conclusion. Google Patents US4241500A - Razor with adjustable blade positioning US4241500A - Razor with adjustable blade positioning ![]() 7/14/2023 0 Comments Spore microscope![]() ![]() Using metagenomic and BLAST analysis, 150 cereal pathogen species (33 different genera) were recorded on the spore trap tapes. , Blumeria graminis, Cladosporium spp., Fusarium spp., Puccinia spp., and Zymoseptoria spp.) were found using these techniques at all sites. Six major cereal fungal pathogen genera ( Alternaria spp. Spore traps were set up in four geographically distinct UK wheat fields (Carnoustie, Angus Bishop Burton, Yorkshire Swindon, Wiltshire and Lenham, Kent). We aimed to investigate the incidence of spores from major fungal pathogens of cereals in the field by comparing microscopic and metagenomic based approaches for spore identification. Fungal diseases such as Fusarium head blight, Septoria tritici blotch, spot blotch, tan spot, stripe rust, leaf rust, and powdery mildew cause serious yield losses in wheat and can impact quality. Wheat is one of the main staple food crops, and 775 million tonnes of wheat were produced worldwide in 2022. 4Institute for Life and Earth Sciences, School of Energy, Geosciences, Infrastructure and Society, Heriot-Watt University, Edinburgh, United Kingdom.3Department of Agriculture, Food and the Marine, Celbridge, Ireland.2School of Biology and Environmental Science and UCD Earth Institute, University College Dublin, Belfield, Ireland. ![]() 1School of Agriculture & Food Science and UCD Earth Institute, University College Dublin, Belfield, Ireland.All rights reserved.Paola Pilo 1, Colleen Lawless 2, Anna M. Fast green Hoescht 34580 Nile red Propidium iodide Spore Staining Wheat germ agglutinin 488.Ĭopyright © 2018 Elsevier Ltd. The proposed staining method has broad applicability and can potentially be applied to follow Clostridium spore behaviour in a range of different environments.ģD-SIM CLSM Clostridium sp. ![]() Direct staining of a cheese specimen using Nile Red and Fast Green allowed in situ observation of spores within the cheese and their position within the cheese matrix. Super-resolution microscopy of spores probed by Hoechst 34580 also allowed chromatin to be visualised. Dual staining with Hoechst/Propidium iodide differentiated spores from vegetative cells, provided information on the viability of cells and was successfully applied to follow spore production induced by heating. Staining was conducted without drying the sample, preventing changes induced by dehydration and cells observed by confocal laser scanner microscopy or using a super-resolution microscope equipped with a 3D-structured illumination module. ![]() Hoechst 34580, Propidium iodide and wheat germ agglutinin WGA 488 were used in combination to stain four strains of Clostridia at different life cycle stages. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pre-treatment. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. ![]() |