Archive for the ‘Microscope Cameras’ Category


Color Infidelity: Love your Monitor!

Stop staring at your screen and give your monitor some love

“It’s like you’re looking right past me, like I’m not even here, and then when you do notice me, all you do is complain, complain, complain. How about a little love, I’m working hard over here!” your monitor screams at you as you question why the image in the microscope doesn’t match the image on the screen. The color accuracy is way off! Maybe it’s the microscope, lighting, or camera. Or maybe the poor color quality is because of your monitor.


Choosing the Wrong Monitor Cheats your Color Accuracy!

When you were considering the microscope options, you bought the best microscope, camera, and software possible with your budget. You were careful; you scrutinized every detail during your decision. Then there, screaming at you toward the bottom of the quote, is that crazy overpriced monitor. You calmly ask your microscope representative: “What do you mean the monitor is $3,000 – I bought one for my computer at home on a Black Friday deal for $150!?!”

Your representative agrees and suggests that you save a few dollars by bringing your own monitor. So you pull one off the shelf, dust it off, and feel proud of yourself for being so thrifty.

Fast forward 30 days, the microscope arrives and is working seamlessly. You’re thrilled, excited, it’s Christmas in February – that is until you’re not happy with the image that you’re seeing. This is NOT the image you saw during the demonstration! The colors are all wrong – but why?


First Impressions are All About the Technology

Not all monitors are created equally and have unique technology driving an image to your eyes. Since we are all technology consumers, we are familiar with some aspects of the technology, but probably not all. Additionally, although we are familiar, we probably don’t fully understand. Below are some of the monitors that’re available on the market and some insight about the technology inside them.

  • CRT
  • Cathode Ray Tube – Big, bulky, low resolution. These fell out of favor in the early 2000’s because of their size – but they were also not great at producing accurate color. This technology would mix red, green, and blue light at a single spot on a screen at a rate of 60Hz. Mixing the three components of light equally should result in accurate color – but unfortunately only about 70% of the color range is displayed.

  • LCD
  • Liquid Crystal Display – The most common form of monitor on the market today. This technology uses an array of liquid crystals which modulate the backlight to produce millions – or billions- of colors.

    The color accuracy depends on the backlight (Halogen or LED) and the display technology (Twisted Nematic vs. In-Plane Switching vs. Vertical Alignment). There are plenty of detailed articles on each; however, it’s safe to assume we have seen the side effects of the Twisted Nematic display where brightness, contrast, and color adjust with the angle of the screen. Recall an occasion where you were adjusting the tilt on your laptop screen and noticed the image changes with the tilt of the screen. In-Plane Switching (IPS) panels have a fewer viewing angle distortions and superior color reproduction (16.7 million vs. 1.07 billion) but are typically more expensive. The Vertical Alignment technology is a hybrid between low cost and accurate color reproduction, but it still can’t quite reach IPS.

  • OLED
  • Organic Light Emitting Diode – These displays feature high pixel density, with individual red, green, and blue LED’s. Color is modulated by adjusting brightness of the three LED’s in one pixel to blend everything together. These are most common on the Android phone or iPhone in your pocket – however they’re making their way into computer monitors, albeit at astronomical prices. OLED’s display 1.07 billion colors like the LCD IPS displays, but are thinner, use less power, and have a better contrast ratio. The color improvement over LCD IPS displays is negligible and not worth the price increase.


    The Ultimate Connection

    Although the importance of the cable connecting your computer to the monitor is often overstated by your local electronics store attempting to sell you a $100 HDMI cable, its important to use the right format.

  • VGA
  • Video Graphics Array – These pesky blue monitor connectors won’t go away. Although your HD monitor works with a 9-pin VGA analog monitor cable, it shouldn’t be used. The VGA cable will distort everything from color reproduction to resolution when it is used. Even black and white images look different with a VGA cable. However, if a digital cable, such as the ones described below, is used, the problems disappear.

  • DVI-I
  • Digital Video Interface – The DVI cable is vastly superior to analog and comes with a variety of pin combinations (DVI-D and DVI-I, Dual link vs. single link). Don’t get caught up in the “more is better” philosophy; any DVI cable with 16 pins will produce an accurate image at 1920×1080.

  • HDMI
  • High Definition Multimedia Interface – Welcome to 19 pins of simplicity. An elegant solution to high quality displays and digital audio. The interface was developed by several companies who charge royalties for it’s use. In addition, there are several different versions of HDMI – so which one do you need? Look for HDMI v2.0 and above and you won’t have any issues with up to 8K resolution or color reproduction. No need for a $100 HDMI cable, the $10 bargain HDMI v 2.0 cable will be just fine, thank you.

  • Display Port
  • Developed as a royalty-free competitor to HDMI, the 20-pin display port cable has all the advantages of HDMI with less confusion about different cable versions. Whether the display port is standard format or mini display port, this format produces up to 8K resolution with no limitations on color reproduction.


    Sooth Your Weary Eyes with Monitor Calibration

    Armed with the information above, you walk into your local electronics store. You’re feeling confident as you look at the monitors, evaluate the technology, compare images from one monitor to the next – side by side – to find one with just the right color. The differences between the monitors are amazing, some images look more saturated, others look cooler or warmer. Is it the difference between a TN display or IPS display? Maybe, but it could also be a simple color calibration.

    Each monitor has “scene selections” and the ability to fine tune brightness, contrast, saturation, and in some cases, gamma. All of these settings can influence the color accuracy of a monitor. A good rule of thumb is to choose sRGB as a scene selection. If that’s not available, maybe pick up an inexpensive monitor calibration device. It sure beats trying to adjust all the monitor display settings manually.


    Resolution, Refresh Rate, Height, Swivel, and Tilt – Do Tell!

    In addition to color reproduction, monitors also differ in resolution, aspect ratio, refresh rate, and ergonomic adjustments such as height, swivel, and tilt. All of the Black Friday deals advertise resolution, so most are familiar with 2K, 4K, and 5K displays – but often overlook refresh rate. Some bargain 4K and 5K monitors only offer that resolution at a paltry 30Hz. At this frequency, even mouse movements appear to lag – 60Hz should be the minimum.

    Another overlooked monitor feature is height, swivel, and tilt. These adjustments can create a comfortable work space and also allow users to adjust the monitor height to fit it into cramped lab space. The bargain bin monitors do not include these adjustments, which are essential to a functional work space.


    Trust W. Nuhsbaum, Inc.

    Technology can be overwhelming and confusing; however, the team at W. Nuhsbaum has selected monitors that will allow investigators to get the most out of their images. The monitors we sell are not the lowest price, but they will deliver the most features for your dollar.

    News Flash: The upstart sCMOS technology is now dominating the market

    Once considered a speed king, sCMOS is showing its versatility

    In the early days of sCMOS technology, manufacturers touted only the readout speed of the sensor. One of the first entry’s into the market, Hamamatsu released the Flash 2.8 with promises of speed – but what it offered in speed it lacked in low light sensitivity. Now with sCMOS sensors offering quantum efficiency of more than 95% the sensitivity issue has been addressed – in force. Although very few sCMOS cameras offer 95% QE, at this point most offer 82% QE.

    In addition to excellent sensitivity, most sCMOS sensors are also extremely low noise. Where most CCD cameras offer readout noise of 6 electrons rms most sCMOS cameras offer a median readout noise of less than one electron rms. The combination of high sensitivity and low noise means that even long exposures provide very clean data for image analysis.


    sCMOS technology takes over for routine imaging

    When sCMOS first entered the market at a price over $20,000, most people still opted for the less expensive CCD technology for any application other than high speed. However, currently, most value oriented sCMOS cameras such as the Leica DFC9000G are available for around $14,000. Some sCMOS camerasare now available for less than $10,000 and creeping into the routing imaging realm.

    The entry in to routine imaging comes with good reason, sCMOS technology is superior to CCD by offering lower noise, higher resolutions, faster readout, and better low light sensitivity. Models such as the QImaging Optimos and pco.edge come in either at or near the $10,000 mark, making sCMOS available for mainstream users.


    sCMOS takes aim at EMCCD

    Earlier in 2016 Photometrics disrupted the EMCCD market by introducing the Prime 95B which offers 95% quantum efficiency and 1.44 million 11 micron pixles with a massive 18.7 mm sensor. The Prime 95B is currently in process of devouring what is left of the EMCCD market. Yes there is still a place for EMCCD for extremely low light applications such as photon counting – but with the Prime 95B the applications are dwindling. Also, with a price that is nearly half of most EMCCD’s, the price advantage is significant as well.


    High Speed in Low Light

    Conventional wisdom in scientific imaging chose EMCCD for applications where high sensitivity and high speed is required for accurate data collection. Again sCMOS technology is challenging conventional wisdom with clever on camera processing.

    The Photometrics Prime employs a process called Prime Enhance where pixel grayscale values are rapidly compared to neighbors thereby reducing the effect of readout noise on the image data. This processing can, in some cases, offer a higher signal to noise ratio (SNR) by a factor of eight! An image acquired at 100ms has equivalent SNR of an image acquired at 800ms!


    The End of Big Data

    When a 4.2 megapixel sCMOS camera is unleashed at 100FPS with 16 bit images, the file size can increase rapidly. Datasets of 100GB are not uncommon in high speed or multidimensional experiments. However, with the release of the Hamamatsu Flash v3 and Photometrics Prime, sCMOS cameras have started to address the issue of file size by offering lower bit depth images and “blank data” elimination with Prime Locate.

    Although disk space is increasingly becoming less expensive, waiting 30 minutes to copy 100GB of data to a network drive is about as exciting as rush hour traffic.


    Trust W. Nuhsbaum

    The world of scientific cameras is rapidly changing with new technology and models released regularly. Trust the imaging experts at W. Nuhsbaum to educate you on the many choices and help select the right camera for your application.

    Love your color microscope camera

    Color infidelity: Why choosing the correct camera is being faithful to your data

    Many microscope users dismiss the color microscope camera as mundane and do not consider the technology with as much thought as a low light monochrome camera for fluorescence. Although this is understandable considering the wide range of options and high degree of pricing variability in monochrome cameras, it is no excuse to dismiss the color camera evaluation process.

    Many disciplines depend on accurate color such as pathology, material sciences, and forensic science – however because of budget constraints or the influence of mainstream technology, users ultimately make decisions based on interface (FireWire, USB 3.0, USB 2.0, Camera Link, etc.), software functions, or the dreaded maximum megapixel count. Although these are important, color accuracy and image quality should sit at the top of the list of items to consider!

    Assuming the user has already addressed Kohler Illumination, lighting conditions, and the objective lens have all been addressed, the camera selected can have major ramifications for image analysis, image quality, and ultimately results.


    It’s Not Flashy, but Your C-Mount Will Help You fall in Love

    In a modern microscope with infinity corrected optics, there is a highly corrected tube lens built into the microscope, which projects the final image to the eyepiece and camera. The final lens is either the eyepieces or the camera c-mount – which matches the microscope magnification to the camera chip size.

    For some reason, unknown to me, c-mounts generally do not have any correction. So if you select a camera with a ½” sensor, there should be a matching 0.5X c-mount – and that convex lens has little to no correction! In theory the sensor uses the center of the lens, which has the best correction, but if someone wants the best image possible they can either use a 1X c-mount with no lens or a 1.1X APO c-mount from QImaging.


    Finding a Heart of Gold in the Specifications

    Many people are familiar with evaluating monochrome cameras based on read noise, pixel size, dynamic range, among other specifications. However, most do not consider these technical specifications when evaluating color cameras. They are critically important to a quality image!

    Generally speaking, inexpensive cameras cut corners on pixel size and noise, which reduce the dynamic range of a camera. These issues will lead to an image that appears grainy with a restricted field of view. Be wary of 1/3 inch sensors and some ½ inch sensors, depending on the number of megapixels, these could lead you down the wrong path.

    Small sensors are contrasted by more expensive cameras, which have larger pixels, are physically larger in area, and are more sensitive. These traits produce a beautiful image with an excellent field of view. Sensors that are 2/3 inch or larger are traits of a quality sensor.


    Beauty is in the Eye of the Beholder?

    When evaluating the output of a color camera, the simple conclusion is that there is a color picture, however, the camera doesn’t have the ability to produce purple, brown, orange, or any other intermediate color without color interpolation. On a digital camera, each pixel is assigned a color, red, green, or blue and based on the signal in neighboring pixels, the camera will produce a color image.

    There are a few ways to address whether the color is correct with lighting, lenses, and alignment, however, the color interpolation algorithms are written by people. Humans. Error-prone people who do their best to faithfully represent color, but are not always perfect. Printers solve this issue with the Pantone color palette, but the same standards do not exist for digital images.

    Users are left to adjust settings such as white balance and saturation to get the image on the computer monitor to represent the view through the microscope eyepeices. This is a highly subjective and error-prone process, because without a color standard such as the one used in ChromaCal, the color accuracy is left to the user – and the quality of the computer monitor. But that’s another blog post…


    Camera Drivers Are Like Tasting the Same Wine in a Different Wine Glass!

    Even with identical hardware components, something as simple as a camera diver can influence color accuracy. Using a QImaging camera in both QCapture Suite and Image Pro Premier will produce different color.

    Color differences between the two programs can, in part, be attributed to subtle differences in default image settings. However, some of the difference is built into the driver itself and depends on the author of the software program and subsequent camera driver.


    Trust W. Nuhsbaum, Inc.

    Since the dawn of digital imaging the microscope sales representatives and imaging specialists at W. Nuhsbaum, Inc. have demonstrated and sold color cameras from companies such as Leica, QImaging, Jenoptik, SPOT, and many more, with several different software programs and hundreds of camera drivers.

    The experience, and customer feedback, regarding color accuracy has provided each sales representative with the expertise to be able to recommend a camera and software package that will most faithfully represent the color in your image. Contact the team at W. Nuhsbaum, Inc. to learn which camera and software platform are best for your application.

    sCMOS runs up the score on CCD

    sCMOS scores a touchdown, spikes the ball, and receives a penalty for excessive celebration

    The technology in complementary metal-oxide semiconductors (CMOS) sensors has been around for a long time. These sensors are inexpensive to manufacture and have made their way into the many imaging devices found in your pockets (phones, cameras, MP3 players, etc.).

    As a technology, CMOS is not merely relegated to funny cat pictures or a video of your buddy’s failed attempt at an American Ninja Warrior course. Instead, CMOS has evolved from a simple sensor design to a niche high-speed camera, all the way to a robust technology that benefits a wide variety of microscopy applications as diverse as time-lapse applications, to cell trafficking, to light sheet microscopy.

    The coming of age of CMOS happened a few years ago with the launch of a new Fairchild sensor design incorporated into cameras such as the Hamamatsu Flash 4 v2, pco.edge, and Andor Neo/Zyla. What has been coined scientific CMOS (sCMOS), in many arenas, has overtaken CCD as the gold standard for fluorescence imaging.

    Although the new generation of CCD sensors has its place alongside sCMOS, as noted in a previous blog post, sCMOS technology has eroded, if not completely replaced, CCD’s position as the preferred sensor technology for advanced imaging applications.


    sCMOS Scores Another Victory in Technological Advancement

    Just in time for the 45th meeting of the Society of Neuroscience in Chicago, IL, Hamamatsu, pco, and Andor have announced major technological advancements in their respective sCMOS cameras. Photometrics announced its entry into the sCMOS market with very unique shot noise reduction technology.

    For Hamamatsu, pco, and Andor, the release of the new cameras include sensors that employ improved microlenses. The microlens improvement increases the overall QE of the sensor by 5-10% across the visible light spectrum. This offers users the ability to capture more light in less time, increasing signal-to-noise ratio, shortening exposure times, and increasing frame rates.

    Although three companies have announced new cameras, only Hamamatsu has access to the new chips now, where other companies will not have access for several more months.

    Without a sCMOS camera to offer microscopists until last week, Photometrics has tossed an 80 yard touchdown-scoring bomb into the sCMOS market. Named the Photometrics PRIME, the standard 4.2 megapixel sCMOS sensor used in the vast majority of cameras in this class has been juiced with noise and data-reducing algorithms. These advanced features are unique and stay with the tradition of advanced technology in flagship cameras from Photometrics.


    Striving for Superior Signal to Noise Ratio

    In low light fluorescence imaging, one of the most important aspects of the detector is signal-to-noise ratio (SNR). Although the equation to calculate signal-to-noise ratio (SNR) is complicated, the concept is simple: How well does the camera sensor read out signal above the level of electronic noise?

    Smart engineers from the various companies have been working on improving SNR and appear to have addressed it in force. The new 82% QE sensors offer an incremental improvement. However, with Photometrics Prime Enhance technology, Photometrics reports an improvement of 3X-5X in SNR. The data provided in Photometrics technical notes provide a glimpse at what is possible. And what is possible is amazing!


    Higher Frame Rates with Fewer Photons

    A characteristic of sCMOS that has always been attractive is high frame rates. If you thought a base, full resolution frame rate of 100FPS was impressive, the Hamamatsu Flash 4.0 v2 can achieve over 25,000 FPS with ROI. These frame rates, however, are exposure-time limited. What is the most common cause of limited exposure time? Photons.

    With a higher QE in the Hamamatsu Flash 4.0 v2 PLUS and the Prime Enhance technology from Photometrics, sCMOS cameras are set to challenge the conventional wisdom that EMCCD’s are required for low light, high speed imaging.

    As documented in the technical note, Photometrics Prime Enhance technology can generate equivalent data in 100 milliseconds for what would normally be 800 milliseconds in standard operation! Take that, EMCCD!

    It is worth noting, however, that although Prime Enhance generates clean and beautiful data, the same cannot be said for the visual image. The Prime Enhance algorithms reduce noise in the pixel gray values but, because noise is reduced by factoring in neighboring pixels, the final result is an image with a Photoshop Palette Knife appearance. This is most noticeable as signal decreases to “near noise” levels, but incredibly, the grey level histogram still looks good. If beautiful images are what you are after, fear not, Prime Enhance can be turned off, exposure time extended, and a beautiful image will result. However, if you want to go fast in low light, Prime Enhance makes it possible!


    More Information with Less Data

    At 4.2 megapixels, 65,536 gray levels (16 bit depth), and 100 FPS, the current generation of sCMOS cameras generates a lot of data! Localization-based super resolution systems are already using sCOMS cameras, which is why the Photometrics PRIME has two more tricks up its sleeve: Prime Locate and Multi-ROI.

    Prime Locate allows the data transfer of only the pixels which register a grey value in localization-based super resolution systems. Considering many of these systems generate 60,000 – 100,000 images before building the journal cover-worthy super resolution image, the data savings will be tremendous. This technology also increases frame rates, lowers file size, and reduces storage concerns.

    The Multi ROI function in the Photometrics Prime also allows users to capture multiple regions of interest (ROI) in a single field of view. So if the user has two small features in one huge field of view, leave the empty data on the microscope and only acquire the ROIs. Reducing file size and collecting more data, what could be better?


    The sCMOS landscape

    With the introduction of new sCMOS sensors and sCMOS sensor technology, the market is rapidly changing. The new 82% QE sCMOS sensors have brought more than just high performance. There are now several variants that differ on price, cooling, triggering, resolution, shutter, interface, and now quantum efficiency.

    While the new Flash 4 V2 Plus is sure to be priced at a premium, companies such as pco have released USB3 cameras at a much lower price, which are based on the current and popular sensor. Whatever your priority, there is a sCMOS camera to step up to the challenge.


    With sCMOS Leading the way, is CCD Obsolete?

    Although sCMOS has firmly positioned itself as the premier technology for high-end fluorescence imaging, it does not cover the entire range of scientific imaging – particularly when price and application are considered. Many investigators are not going to be able to justify the price premium on sCMOS cameras for features that will be rarely used on, for example, a routine fluorescent stereo microscope.

    Although CCD has fallen out of favor for high-end widefield acquisition, it still has its place on microscopes. Recently QImaging released the Retiga R1, R3, and R6 cameras at shockingly low prices. With the Retiga EXi, what used to cost $12,000 three months ago has bottomed-out at less than $5,000 with the introduction of the new Retiga R1. In addition, the new R1 has deeper cooling, higher QE, and a higher frame rate for live cell imaging!

    Interestingly, this opens the door to simultaneous, multi-channel, imaging applications that require several cameras and employ the Multi-Cam from Cairn Research. What used to be a $40,000-$60,000 investment now costs a fraction of the price because of QImaging’s new CCD cameras!


    Trust W. Nuhsbaum, Inc.

    Choosing a camera can be intimidating, but identifying the needs for the application is the first step in making a smart decision. When evaluating technology for your lab, which will be used for many years into the future, it’s important to consider the latest products and technology advancements.

    The Imaging Specialists at W. Nuhsbaum, Inc have seen cameras evolve over the years and can provide perspective on the latest technology to arrive on the market. Trust the experience of W. Nuhsbuam, Inc to weather the technology winds of change and advise on the proper technology for your experiments.

    Megapixel meltdown: Canon’s 250 megapixel sensor

    Megapixel Meltdown: Cannon announces the release of a 250 megapixel sensor

    With the recent release of a 250 megapixel sensor, the world of microscopy will never be the same. Or will it? Although microscope users will continue to demand higher resolution cameras it is not the primary factor for comparison.

    For years consumer electronics have lead the way in educating people in the standards for how to evaluate a camera. The most common standard – megapixels. Consumer camera manufacturers make little to no mention of the lens, resolving power, sensor noise, dynamic range, sensitivity or anything that actually influences quality images. So when Canon releases a 250 megapixel sensor, consumers immediately assume more is better – and that sensor should be connected to the microscope!

    Although the 250 megapixel sensor will allow an investigator to make a high resolution print the size of a small house, it will not allow said investigator to learn more about the sample in the image.

    Resolving power of optical system influences the need for sensor resolution

    Although camera sensors are amazing at capturing the detail in an image provided by a series of lenses, the series of lenses dictate the resolving power of the system. In short, unless the optical system is capable of resolving to the level of the camera pixel, the camera’s pixels are wasted. There could be more than one pixel capturing the same exact feature on the sample, providing your image a lot of pixels, but not a lot of data. Thankfully smart people who know numbers have worked out the details of digital sensors in microscopy. Some important equations are below:


    Optical Resolution (um) = (0.61 * wavelength)/(NA)

    Object size (um) = (Optical Resolution * Objective Magnification (i.e. 63X)/(c-Mount Adaptor)

    Nyquist Sampling Frequency: Object size/2.3 = Pixel Size to Resolve Object (um)

    Alternatively, if one would simply prefer plug numbers into a calculator, there is a web based calculator available also. Or for more information, Photometrics has an excellent learning zone dedicated to matching resolution.

    In the case of the Canon sensor, the pixels are about 1.49 microns across, so in the case of a compound microscope with a 10X objective lens with a numerical aperture of 0.4 and a lens free 1X c-mount, the ideal pixel size would be 3.646 microns. Or in the case of a stereo microscope with a 1X objective lens ideal pixel size would be about 3.5 microns.

    At 1.49 microns, the 250 megapixel sensor is at least 2X over sampling at low magnification and 8X over sampling at high magnification. Furthermore, this behemoth sensor is so big (29.2 x 20.2 mm) that it would require a magnifying intermediate lens to avoid vignetting – making the pixel size over sampling problem worse.


    Microscopes are made for imaging molecules, not mountains

    Nothing is more awesome than one upping a friend as it relates to Megapixels. If this were not true, Apple would not have released an iPhone with 4K streaming and a 12 megapixel sensor. Nothing like megapixels to make someone feel like their phone is obsolete…

    From the calculations above it’s clear that there is a limit to the value of megapixels as it relates to resolution. In the case of the 250 megapixel sensor, the developers never intended for the sensor to go onto a microscope, since they cite the sensors ability to read a serial number off an airplane from 11 miles away. Ideally Canon’s 250 megapixel sensor would be used for telescopes, not microscopes.

    When considering the specialized optics of light microscopes there are practical limits to resolution. Cameras that go beyond the three micron pixel size limit are simply offering users bragging rights, because there is not benefit to additional data.


    Real factors to consider when evaluating a camera

    While megapixels are a simple number to digest and compare across many different products, there are other factors to consider:


  • Quantum Efficiency

  • Read noise

  • Signal to noise ratio

  • Full well capacity

  • Dark current

  • Pixel size

  • Sensor size

  • All of these factors contribute to what many people will consider to be a beautiful image. Inky blacks with vibrant whites all while providing the field of view and resolution that is enough to record data.


    Trust W. Nuhsbaum, Inc.

    Choosing a camera can be intimidating, but the process can be made easy by consulting with the team of microscope and imaging specialists from W. Nuhsbaum, Inc. Trust the experience of W. Nuhsbuam, Inc to weather the technology winds of change and advise on the proper technology for your experiments.

    Sony announces CCD production to end

    Sony announces end of life for CCD production: The CCD zombie apocalypse is upon us!

    Just in time for the mid-season restart of AMC’s The Walking Dead, on January 30th 2015 Sony let the cat out of the bag that they would stop production of its existing line of CCD sensors. There is a difference, however, in announcing the end of production, and actually ending production. Current reports suggest that the end will not actually happen for another 10 years.

    When the end does finally arrive, unlike the zombies popularized in The Walking Dead, CCD cameras will not die and come back as half dead creatures. Cameras that employ CCD sensors will continue to function happily for years to come – with new cameras built on CCD technology ready to release now, and into the future.


    The Future of CCD sensors

    While many might assume Sony’s announcement signals the end of life for CCD sensors, there will be plenty of time to prepare. Although the rumor mill is inaccurate with dates, there are reports that Sony targeted final ship dates for 2025. As of today, 2025 is a full 10 earth years away. Or in technology years, approximately 2,200 – that’s a joke people.

    Unfortunately there is no public announcement from Sony regarding the end of CCD’s, however there are CCD distributors that made the announcement for them.

    Thankfully there are other companies out there that produce CCD sensors who will be happy to pick up the slack provided by Sony – such as Texas Instruments. There are many other, smaller companies that may pick up the slack as well which can be found on an unconfirmed list of CCD manufacturers.


    Impact of Sony’s discontinuation of CCD’s for scientific cameras

    In the short term, there is little impact. With years to prepare, camera manufacturers can continue using the wildly popular Sony ICX-285 sensor cameras such as the Photometrics HQ2, Leica DFC365, and Qimaging QIClick.

    In addition to the continuation of existing camera models, companies such as QImaging are actually preparing the launch of a new camera line which includes Sony sensors. The all new QImaging R1 camera uses the successor to the Sony ICX-285 chip, the new ICX-825 sensor. With and improved quantum efficiency of 75%, 16 bit digitization, higher frame rates, the QImaging R1 is betting on the availability of Sony CCD’s for years to come.

    Interestingly, Hamamatsu may also be prepared to pick up the demand for scientific cameras based on CCD technology as it owns the technology in its ER-150 sensor. Since Hamamatsu does not make public the company they use for production, with the many manufacturers of CCD sensors, one must assume the Hamamatsu ER-150 is safe.

    Hamamatsu has kept its technology secret for years and with the ER-150 sensor being used in the Hamamatsu Orca R2, Orca D2, and 8484 series of cameras, Hamamatsu is uniquely positioned to meet the demand for scientific cameras built on CCD technology.


    Expected lifetime of CCD’s in scientific cameras

    If the CCD sensor built into your favorite camera is working well now, chances are it will work well for years to come. Particularly when purchased from an established manufacturer such as the brands represented by W. Nuhsbaum, Inc.

    Many of the first generation monochrome CCD sensors are still being used today in many scientific labs and Research and Development facilities around the country. Although the Hamamatsu Orca 100 was discontinued many years ago, there are plenty of 15 year old Hamamatsu Orca 100 cameras that still function well for low light fluorescence imaging.


    Buying technology with a defined end date

    Although purchasing a camera with a component that is slated for cancellation may be intimidating, consider that hundreds of components go into scientific cameras – any of which could be changed with very little notice. Sony is doing everyone a favor by making the end of life clear to manufacturers so they can prepare.

    The current time frame of 10 years is very, very long particularly in the technology sector that develops so quickly. There are numerous cases of technology becoming obsolete after 10 years not because of hardware or electronics inside of a device, but because of software support, driver support, data transfer interface, or a new innovation that supplants the existing technology. Although the device may still work, it is unusable because of something as silly as computer manufacturers making a switch from 5V PCI bus to 3.3V PCI bus.


    Trust W. Nuhsbaum, Inc.

    Choosing a camera can be intimidating, but identifying the needs for the application is the first step in making a smart decision. The second step is to disregard rumors and big announcements, and instead focus on the image and application. If your application is best addressed with a CCD sensor, fear not, CCD’s will be around for a long time. Although they will go away some day, it will not resemble a CCD zombie apocalypse where all CCD’s die and come back as half dead creatures.

    Trust the experience of W. Nuhsbuam, Inc to weather the technology winds of change and advise on the proper technology for your experiments.

    sCMOS v CCD – adversaries or partners?

    Scientific CMOS cameras: Arch rivals to CCD or partners in crime?

    When building a microscope system, there are many important components to consider, such as the microscope stand, objective lenses, and contrast methods. However, increasingly software and cameras are playing a critical role in the design of new microscope systems. At the forefront of this paradigm shift are the many new university faculty members setting up new labs, making decisions that will affect the next several years of their research projects. Therefore, considering the investment required for a quality camera and software package, it is imperative to get the decision right – the first time.

    While every point in time has new technology to consider, the decision facing new investigators regarding fluorescence microscope cameras has never had two such strongly opposing forces as it does today. Researchers are accustomed to opposing technology for color and monochrome fluorescence microscope cameras, but now there is a dramatic difference in fluorescence microscope cameras – Charge Coupled Device (CCD) vs. scientific complementary metal oxide semiconductor (sCMOS).

    In the past, the difference was cooling, length of exposure, and resolution. Today, the decision includes frame rate, noise, resolution, bit depth, gain, binning, dynamic range, and quantum efficiency. These many variables can make even the most tech-savvy scientist’s head spin. When considering a microscope system, new investigators can feel the tension between the two technologies. Adding to the pressure is the sense that one is buying “old” technology if choosing a CCD microscope camera instead of the newer sCMOS – but are they really opposing forces where sCMOS will replace CCD for monochrome fluorescence cameras? Or are they two independent technologies that work together to cover the wide range for fluorescence microscopy applications?


    Applications that determine the sCMOS or CCD microscope camera


    High speed

    When looking for pure speed, it’s hard to find a camera for fluorescence imaging that beats sCMOS. There are some electron multiplying CCD (EMCCD) cameras that with region of interest (ROI), binning, and unreasonable clock and clearing settings that can challenge the raw speed of the sCMOS camera, but at 100 FPS full frame and 25,000 FPS or higher with ROI, the sCMOS camera is king of speed. By contrast, CCD sensors are slow – most topping out around 100-150 FPS with ROI and binning. So not only is the camera slower, but to get to the base line speed of sCMOS, the user needs to compromise on resolution and field of view.

    Winner: sCMOS

    High speed in low light fluorescence

    Speed is where the clear victory ends, because when considering frame rates, one must also consider exposure times. So unless your exposure times are on the microsecond time scale, stop dreaming about 25,000 FPS. However, at 10 milliseconds, 100 FPS is at least plausible. Although 10 ms is a very short exposure time for fluorescence, the 16 bit register and 65,536 grey levels means the sensor can produce significant grey levels above background noise. Combined with the lower read noise in sCMOS the user gets more of what they need: pixel data.

    The CCD sensor, surprisingly, has some virtuous characteristics that benefit users in low light conditions, most notably analog gain and binning – two features that are unavailable on sCMOS cameras. These two functions, although they amplify noise and reduce resolution, provide the user with the ability to amplify signal by multiplying the electrons produced by the sensor and create “super pixels” – effectively increasing the sensitivity of the sensor. These two functions, combined with a 12-, 14-, or 16-bit digitizer with 4,096, 16,384, or 65,536 grey levels respectively, can amplify signal and get what users need most: pixel data.

    Winner: Tie

    Multi dimensional live cell

    Live cell imaging presents its own challenges, although live cell could require both speed and low light sensitivity, a lot of live cell is slow, time lapse imaging. Long exposure times? Who cares! There are 10 minutes between each image.

    As far as field of view, sCMOS has a 50% larger field of view over most fluorescence CCD cameras. This means each position can increase sample size and possibly decrease the number of images required to collect the necessary data. When smaller areas are required, the user can simply add an ROI and create a mosaic image with smaller images, albiet larger than the single image FOV.

    Another area for importance is sensitivity (QE), noise, and the resulting signal to noise ratio. Whenever real data is required for image analysis, more signal and less noise is always preferred. With readout noise around 1 electron rms, the sCMOS wins the day compared to readout noise of 6 electrons rms found in the most popular CCD sensors.

    Where CCD provides a counter punch is with dark current noise which can range but is normally on the order of 0.0002 electrons/pixel. By contrast, the industry leading Hamamatsu Orca Flash 4 comes in at 0.06 electrons/pixel. This difference is orders of magnitude in favor of the CCD, however, the dark current noise doesn’t present itself until exposure times go out into multiple seconds to minutes of exposure. And really, who wants to wait that long for an image particularly if there are multiple channels, Z stacks, and stitching taking place.

    Winner: sCMOS

    Deconvolution

    Deconvolution requires proper digital imaging sampling rates, pioneered by Harry Nyquist, which is the topic of an article unto itself. However, the good news is that both CCD and sCMOS have an excellent pixel size for proper deconvolution with 63X and 100X lenses.

    Most CCD sensors have a 6.45 micron pixel size, sCMOS have a 6.5 micron pixel size – which is perfect. However, CCD sensors produce smaller data sets which when working through the calculation heavy deconvolution algorithms dramatically speed the time to result. Although the larger file size of the sCMOS takes time for deconvolution, the end result is completely stunning. The high resolution 4.2 megapixel image with zero light scatter is simply unbeatable.

    Winner: Tie

    Mosaic Image Stitching

    When capturing a large area with a high magnification, the only method for acquisition is mosaic image stitching. In this application field of view equals speed, and the sCMOS sensor has a 50% larger field of view, which means 50% fewer images and 50% more speed. When a CCD is used in combination with Objective Imaging Surveyor software and Turboscan, the CCD can go faster, but not as fast as the sCMOS and its huge field of view.

    Winner: sCMOS

    Ratiometric Imaging (FURA and FRET)

    When comparing the difference between image 1 and image 2, ratiometric data is only as good as the grey level difference and time in between the two images being compared. One might assume higher frame rates are better, however, frame rates are dictated by the biological process they are measuring. In this case, biology is the neutralizer because FRET and FURA measures fluorescent proteins or calcium dyes, not extremely bright quantum dots. So attributes such as a high bit depth, gain, and binning can be very helpful. However, this needs to be balanced with speed and all of that depends on the experimental biology. In this case, there is no clear winner.

    Winner: Tie

    Quantitative fluorescence

    If you were to ask 10 people involved in biological imaging to explain quantitative fluorescence microscopy you would probably get three or four answers that focused on different concepts. Comparing fluorescence intensity, ratiometric imaging, and photon counting would probably arrive at the top of the list. Your journey through the world of quantitative fluorescence microscopy would begin there and you would embark on a long voyage, for which a camera would naturally be required.

    Important characteristics for a quantitative fluorescence microscopy camera would be low read noise, good full well capacity, high dynamic range, and a high bit depth. All of which sCMOS cameras have. For example, the pco.edge 4.2 has a read noise of 0.8 electrons, dynamic range of 37,500:1, full well capacity of 30,000 electrons, and a 16 bit digitizer. By contrast, CCD cameras such as the Hamamatsu Orca R2 have a read noise of 6 electrons, dynamic range of 3,000:1, full well capacity of 18,000 electrons, and two digitizers of 12 and 16 bits. If the numbers look intimidating, sCMOS is bigger, and in this case, bigger is better.

    In the case of quantitative fluorescence microscopy, sCMOS is truly the better fit. Ratiometric imaging depends on the biology, so there is no clear winner. If one is interested in photon counting, welcome to the land of EMCCD cameras, where signals are low and camera sensitivity is high.

    Once the camera is selected, the method for qualifying fluorescence begins. Expanding on this part of the journey is best left to highly experienced scientists. A great resource for learning more about this method is from the book Quantitative Imaging in Cell Biology: Methods in Cell Biology by Waters and Wittmann.

    Good luck on your journey!

    Winner: sCMOS

    Trust W. Nuhsbaum, Inc.

    Choosing a camera can be intimidating, but identifying the needs for the application is the first step in making a smart decision. The second step is to not get caught up in any trend or hype around a specific technology. The differences are clear, but although there is some overlap in technology, both sCMOS and CCD have their place in fluorescence microscopy. So although they oppose each other on occasion, in many situations they compliment each other under the large umbrella of fluorescence microscopy.

    The Imaging Specialists at W. Nuhsbaum, Inc have been around long enough to know about where each technology fits and can advise on what is best for your set of applications. Trust the experience of W. Nuhsbuam, Inc to weather the technology winds of change and advise on the proper technology for your experiments.