- Improved Lateral Resolution of 170nm
- Better Signal to Noise Ratio and High QE
- Open System Architecture
- Deeper Confocal Z-Sectioning
- High Sensitivity and Low Noise
- Aﬀordable Budget
During scanning, re-scan mirrors (SM2) move faster than the first scan mirrors (SM1), which magnifies the image on the camera chip compared to the sample, and eventually results in the higher resolution of the image. The resolution of the system can be improved with the re-scan step by a factor of √2 (i.e. 1.41 times), compared to Abbe’s resolution limit by changing the angular amplitude of the re-scanner (SM2). Reduction of pinhole is no longer necessary to increase resolution – in fact, closing down the pinhole will only limit the amount of light passing through, and increase the signal to noise ratio due to weaker signal. Since the re-scan is purely optical method, and no further image processing is required, there is also no time cost of improving the resolution.
The excitation lasers (blue and yellow lines) are directed via a dichroic mirror towards the first scanning unit SM1. As in a standard confocal microscope, the scanning unit scans the laser light in the sample and de-scans the emission light, directing it at the pinhole PH (green and red lines). After the pinhole, a second re-scan unit SM2 directs the light onto a camera chip.
Because of the sensitive camera as a detector, instead of the photo-multiplier tube as in most confocal microscopes (except the spinning disc systems), the signal-to-noise ratio of the RCM is 4 times higher than in standard confocal microscopy.
As described in “Re-scan confocal microscopy: scanning twice for better resolution; G. M. R. De Luca, R. M. P. Breedijk, E. M. M. Manders et al. Biomedical Optics Express 4, 2644-2656 (2013)”, this improved resolution is pinhole independent and therefore a strongly enhanced photon-eﬃciency (and lower photo-toxicity) is observed. Additional deeper confocality in the sample is established.
In the RCM unit, the excitation path is similar to traditional confocal systems (the blue light path in the image above). The unique feature of the RCM is based on the additional re-scanning mirrors used for the emission ﬂuorescence (the green light path above), which is written on a sensitive camera chip. Together with the open pinhole (AU=2), this provides an exceptional image with high resolution and superior signal to noise ratio. The lateral resolution of the RCM is increased to 170nm compared to 240nm of regular confocal microscope and the signal to noise ratio is 4x better.
Multi-channel image and 3D reconstruction with RCM.
|Lasers||Support up to 4 laser lines: 405, 488, 568, and 638nm|
|Additional laser wavelengths upon request|
|3rd party laser combiner supported|
|Laser output through single line fiber (FC Connector)|
|Microscope||Standard microscope with C-mount adapter|
|Major brand microscopes supported|
|Camera||Hamamatsu ORCA-Flash 4.0 (V2 or V3)|
|Andor iXON 897|
|PCO Edge 4.2|
|Photometrics Prime 95B|
|Other major EMCCD/sCMOS camera supported (min 6.5um pixel size)|
|Features||170nm lateral resolution (FWHM @ 488nm)|
|600nm axial resolution (@488nm excitation)|
|1fps (@ 512 x 512)|
|50um (2.2 AU for 100x objective)|
|Optimized for 100x, 60x and 40x objectives|
|Software||Image acquisition software included|
|Other major image processing software packages supported|