“Lightsheet Readout Mode” is a unique and patented feature of Hamamatsu sCMOS cameras which can improve signal to noise ratios in Lightsheet microscopy.
Utilizing selective plane illumination, Lightsheet microscopy is a fluorescence method that places a thin sheet of excitation light across only the focal plane being imaged. The sheet of light is swept in very precise synchronization to the rolling shutter of the camera. The scan height of the rolling shutter effectively becomes a slit, enabling less background signal and scattered light. By using this combination of illumination and camera readout, very fast acquisition of optically sectioned images is enabled. Additionally, there is significantly reduced photobleaching and phototoxicity to the sample as compared to many other methods.
Lightsheet microscopy is used for live cell imaging at scales that range from small groups of cells to whole organisms. The rapid increase in the use of this technique over the past several years has been remarkable.
Exposure and readout methods of CMOS image sensors are classified into two types; Rolling shutter and global shutter. Rolling shutter uses sequential exposure and readout with one pixel or one line as a single unit. Thus, the exposure timing is different within one image capture. Global shutter performs exposure and readout of all pixels at the same time, the exposure timing within one capture is simultaneous. (See Fig 1.)
To take advantage of the nature of the sweeping light in this application Hamamatsu Photonics K.K. developed Lightsheet Readout Mode using the intrinsic characteristics and speed of rolling shutter readout.
Fig 1. Readout timing of rolling shutter and global shutter
The standard mode of operation in sCMOS cameras is to readout each line from the center of the sensor outwards upper half and lower half simultaneously, row after row. (See Fig 2.)
With Lightsheet Readout Mode, the user can scan the sensor in one continuous sweep. (See Fig 3.)
Fig 2. Standard mode
Fig 3. Lightsheet Readout Mode
Lightsheet Readout Mode provides selectable rolling shutter operations from top to bottom and bottom to top readout. (See Fig 4. and Fig 5.)
Fig 4. Top to bottom readout
Fig 5. Bottom to top readout
The rolling shutter mechanism used in Lightsheet Readout Mode enables exposure reset right before exposure of the next line starts. This feature provides sharper and more resolved images by eliminating scattered light.
By comparison, using global shutter all of the pixels are exposed simultaneously and scattered light from the illuminating lightsheet can fall onto neighboring lines, causing blurred images. (See Fig 6. and Fig 7.)
Fig 6. Exposure reset
Fig 7. Effect of exposure reset
“The gentle illumination schemes of light-sheet microscopes allow for imaging thick samples with unprecedented speed and resolution. However scattering within the sample can impair image quality. Electronic confocal detection in modern sCMOS cameras allows for rejection of scattered photons, resulting in better image quality."
Lars Hufnagel, PhD
In order to synchronize a variety of laser widths and scanning speeds with the sensor, Lightsheet Readout Mode offers a flexible readout interval and the ability to set exposure times independently. The user can easily adjust the readout interval from 10 us to 100 ms for each line. (See Fig 8.)
Fig 8. Flexible adjustment of readout interval and exposure time
External trigger output has been built into Lightsheet Readout Mode whereby the camera generates an external trigger along with each line or frame, which enables the user to control peripheral devices such as a stage or light source. (See Fig 9.)
Fig 9. External trigger output
|Japan||Patented||JP06475307, JP05770958, JP06240056, JP05639670|
GB2544913, GB2544666, GB2544667, GB2535367, GB2532906, GB2531200, GB2524183, GB2523012, GB2523263, GB2522793
|Deutschland||Patented||DE112014006152, DE112014000240, DE112014000195.3|
|United States||Patented||US10142566, US9423601, US10362245, US10602087|
|China||Patented||ZL201480002237.1, ZL201810113454.8, ZL201810113455.2|
It looks like you're in the . If this is not your location, please select the correct region or country below.
You're headed to Hamamatsu Photonics website for JP (English). If you want to view an other country's site, the optimized information will be provided by selecting options below.
For modern websites to work according to visitor’s expectations, they need to collect certain basic information about visitors. To do this, a site will create small text files which are placed on visitor’s devices (computer or mobile) - these files are known as cookies when you access a website. Cookies are used in order to make websites function and work efficiently. Cookies are uniquely assigned to each visitor and can only be read by a web server in the domain that issued the cookie to the visitor. Cookies cannot be used to run programs or deliver viruses to a visitor’s device.
Cookies do various jobs which make the visitor’s experience of the internet much smoother and more interactive. For instance, cookies are used to remember the visitor’s preferences on sites they visit often, to remember language preference and to help navigate between pages more efficiently. Much, though not all, of the data collected is anonymous, though some of it is designed to detect browsing patterns and approximate geographical location to improve the visitor experience.
Certain type of cookies may require the data subject’s consent before storing them on the computer.
This website uses two types of cookies:
There are two ways to manage cookie preferences.
If you wish to restrict or block web browser cookies which are set on your device then you can do this through your browser settings; the Help function within your browser should tell you how. Alternatively, you may wish to visit www.aboutcookies.org, which contains comprehensive information on how to do this on a wide variety of desktop browsers.
Occasionally, we may use internet tags (also known as action tags, single-pixel GIFs, clear GIFs, invisible GIFs and 1-by-1 GIFs) at this site and may deploy these tags/cookies through a third-party advertising partner or a web analytical service partner which may be located and store the respective information (including your IP-address) in a foreign country. These tags/cookies are placed on both online advertisements that bring users to this site and on different pages of this site. We use this technology to measure the visitors' responses to our sites and the effectiveness of our advertising campaigns (including how many times a page is opened and which information is consulted) as well as to evaluate your use of this website. The third-party partner or the web analytical service partner may be able to collect data about visitors to our and other sites because of these internet tags/cookies, may compose reports regarding the website’s activity for us and may provide further services which are related to the use of the website and the internet. They may provide such information to other parties if there is a legal requirement that they do so, or if they hire the other parties to process information on their behalf.
If you would like more information about web tags and cookies associated with on-line advertising or to opt-out of third-party collection of this information, please visit the Network Advertising Initiative website http://www.networkadvertising.org.
We use third-party cookies (such as Google Analytics) to track visitors on our website, to get reports about how visitors use the website and to inform, optimize and serve ads based on someone's past visits to our website.
You may opt-out of Google Analytics cookies by the websites provided by Google:
We inform you that in such case you will not be able to wholly use all functions of our website.