Detailed | sCMOS camera high-speed imaging performance in the field of biological applications? Look and know

Help users quickly find samples and adjust focus

The first benefit of high-speed imaging is that it is more convenient to find samples. In the "old ccd era" where the camera's imaging speed is still very slow, in order to quickly adjust the focal plane position and find suitable samples, the experimenter must observe directly under the eyepiece. This is not difficult for brightfield imaging, but some fluorescent signals may be too weak to be directly observed under the eyepiece, which gives the experimenter operational inconvenience.

The high sensitivity of flash4.0 makes it possible to image the sample with low exposure time under weak fluorescence. The experimenter can directly observe the sample under the microscope and adjust the stage position and image focal length without “delay”. Under reasonable conditions, the excitation fluorescence power is greatly reduced to minimize phototoxicity.

This feature makes it unique in some special applications. For example, patch clamps, which require the avoidance of mechanical vibrations as much as possible, enable high-speed cameras to microscopically manipulate neurons with glass electrodes on the display screen (to avoid the shock caused by the naked eye under the eyepiece).

Application conditions: Real-time display without delay requires a camera of 30fps or more

Product model recommendation: flash4.0 lt+/flash4.0 lt

Capture high-speed moving biological samples

Some biological samples undergo rapid displacement changes during the observation process. Sometimes it is necessary to record these fast-moving samples in detail. The faster the frame speed that the camera can reach, the clearer the depiction of these displacement details.

Original video point here >>

Mouse cardiomyocyte beat high-speed imaging 100 frames / sec

Application conditions: Brightfield high-speed imaging requires only a high-speed scmos camera, and the heart-related imaging speed is generally above 100fps.

Product model recommendation: flash4.0 lt+ / flash4.0 lt

Original video point here >>

Zebrafish blood vessel high-speed fluorescence imaging 400 frames / sec slow down 20 times

Application conditions: High-speed fluorescence imaging requires a highly sensitive and high-speed scmos camera. The observation of fast-flowing fluorescent markers requires more than 200fps, and the camera qe needs more than 80%.

Product model recommendation: flash4.0 v3

Capture rapid changes in fluorescent signals

Many physiological and biochemical processes are accompanied by rapid changes in fluorescent signals. These fast-changing signals are sometimes indistinguishable by the naked eye. At this time, the high-speed imaging of flash4.0 can capture this information well.

Ultra-high speed fluorescence imaging of neuronal membrane potential

Application conditions: High-speed imaging of membrane potential as a special application, only sCMOS cameras can be used. The observation of fast-flowing fluorescent markers requires more than 200fps, and the camera QE needs more than 80%.

Product model recommendation: Flash4.0 V3

In the above figure, the fluorescence signal of the neuronal cell body due to the change of membrane potential is not visible to the naked eye; however, after the time analysis of the gray level of the image under high-speed imaging, the fluorescence in the 1 ms time scale can be seen quickly. Changed.

Other changes that can be observed by the naked eye, through high-speed imaging, can better study the change of its signal over time. For example, calcium ion fluorescent dyes or fluorescent proteins (fluo-4, gcamp, etc.), the fluorescence intensity increases with the increase of calcium ion concentration.

Cardiomyocyte calcium ion concentration changes with time 200 frames per second slows 10 times

Application conditions: Fluorescence high-speed imaging requires a highly sensitive and high-speed scmos camera. The observation of calcium wave and calcium spark requires more than 200fps, and the camera qe needs more than 80%.

Product model recommendation: flash4.0 v3 / flash4.0 lt+

Signal transduction and downstream effect recording

The high speed of flash4.0 can also record the transduction of biological signals and the downstream effects of cell production below the millisecond level.

For example, in the following example, after the simultaneous imaging of the cell phase calcium signal and the cell systolic diastolic phase difference, quantitative analysis of the grayscale change of the image can obtain time information of two events. Comparing the two time curves, we can see that the calcium ion signal changes first, then the cell undergoes contraction deformation, and the time difference between the two is the calcium ion signal transduction time.

Apoptosis of cardiomyocytes induced by ips and changes in calcium signaling

Application conditions: Fluorescence high-speed imaging combined with bright field for simultaneous imaging requires a highly sensitive and high-speed scmos camera, and with a two-color splitter component, the machine needs more than 200fps, and the camera qe needs more than 80%.

Product model recommendation: flash4.0 v3/ flash4.0 lt+ + w-view gemini

With the application of high-end imaging methods

The rapidly developing microscopic imaging technology is constantly placing new demands on the performance of photodetectors. In the depth imaging of thick samples or super-resolution imaging of subcellular structures, flash4.0 cameras have a place in the field of imaging detectors with their superior performance.

1. Light-sheet illumination has significant advantages in the imaging of animal organs and embryos in some models because of its z-axis resolution comparable to laser confocal and faster imaging speed than laser confocal. The high frame rate imaging capability of the flash4.0 camera, combined with the scanning of flake light, enables rapid z-stack scanning of thick samples and reconstruction of their 3d structure. This method is especially suitable for taking pictures of high-speed changes in tissues and organs (such as heartbeat).

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2, spinning disk confocal as a very mature confocal imaging means, can provide point scanning confocal unmatched high-speed imaging capabilities. In addition to the multi-pinholes brought by the nipkow turntable, this high-speed imaging capability requires the high-speed shooting performance of the scmos to achieve a high-resolution image equivalent to point-scanning confocal video-rate.

3. The great breakthrough achieved by super-resolution imaging technology in recent years has caused humans to break the diffraction limit of visible light and increase the sub-microscopic resolution of cells in the visible light range to a level comparable to that of electron microscopy.

Among the various super-resolution technologies, flash4.0's high-speed and high-resolution chips are well-suited for use in a large class of super-resolution imaging called smsn (single-molecular switching nanoscopy) (including our well-known strom and palm). Through a short period of random single-molecule fluorescence on-off, a super-resolution image is finally formed on the millimeter scale time scale. It is the high speed and high sensitivity of flash4.0 that allows the experimenter to use a higher frequency to randomly excite the single-molecule fluorescent switch, thereby shortening the imaging time of the entire system.

Attachment: The relationship between the frame rate of the flash4.0 camera and the size of the roi

Ok~flash 4.0 camera Jun's explanation is here, because the platform video restrictions, there may be, you want to see the original video of the small partner, you can scan the QR code to see the original ha ~

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